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Volume 18,Issue 2,2011 Table of Contents
2011, 18(2):115-125.
DOI: 10.3872/j.issn.1007-385X.2011.2.001
Abstract:
With the fast development of early diagnosis and treatment for cancers, the survival rates of victims of this deadly disease has been greatly increased. Yet the survivors still face the threat of reoccurrence. More and more evidence documented that these disease-free patients (and even some so-called healthy people) harbor dormant tumor cells, which are now regarded as the main cause for tumor reoccurrence. Dormant tumor cells refer to a group of cells featured by its minimal number and the inconvenience for detection. These cells are caught in cell cycle arrest and can exist in both patients and healthy people. At present, the mechanism of how these cells are formed and dormancy timing patterns are still unknown. As the bone marrow and peripheral blood of patients are easy obtained, sensitive detection method has been established aimed to detect dormant tumor cells from these two sources. Evidence has shown that immunological suppression, angiogenesis in tumor tissues as well as surgery may all play roles in tumor dormancy activation, and thus lead to tumor reoccurrence and metastasis. As a result, dormant tumor cells have become a crucial target for preventing and treating cancers. However, since dormant tumor cells stay in a tranquil state, they are insensitive to traditional clinical therapies including radiation treatment and chemotherapy, and therefore impose great difficulty for the radical cure. According to the latest research, immune surveillance is closely related to the phenomena of tumor dormancy. As a result, adoptive immunotherapy may bring radical revolution in the area of tumor prevention and treatment. We hope that, with the deepening of research on tumor dormancy, clinical options with high efficiency will be available to wipe out tumor cells or induce tumor cells into permanent dormancy.
With the fast development of early diagnosis and treatment for cancers, the survival rates of victims of this deadly disease has been greatly increased. Yet the survivors still face the threat of reoccurrence. More and more evidence documented that these disease-free patients (and even some so-called healthy people) harbor dormant tumor cells, which are now regarded as the main cause for tumor reoccurrence. Dormant tumor cells refer to a group of cells featured by its minimal number and the inconvenience for detection. These cells are caught in cell cycle arrest and can exist in both patients and healthy people. At present, the mechanism of how these cells are formed and dormancy timing patterns are still unknown. As the bone marrow and peripheral blood of patients are easy obtained, sensitive detection method has been established aimed to detect dormant tumor cells from these two sources. Evidence has shown that immunological suppression, angiogenesis in tumor tissues as well as surgery may all play roles in tumor dormancy activation, and thus lead to tumor reoccurrence and metastasis. As a result, dormant tumor cells have become a crucial target for preventing and treating cancers. However, since dormant tumor cells stay in a tranquil state, they are insensitive to traditional clinical therapies including radiation treatment and chemotherapy, and therefore impose great difficulty for the radical cure. According to the latest research, immune surveillance is closely related to the phenomena of tumor dormancy. As a result, adoptive immunotherapy may bring radical revolution in the area of tumor prevention and treatment. We hope that, with the deepening of research on tumor dormancy, clinical options with high efficiency will be available to wipe out tumor cells or induce tumor cells into permanent dormancy.
ZHENG Qiu-hong
,
XU Yang-mei
,
WEI Zhi-qiang
,
GONG Fu-sheng
,
YANG Jian-wei
,
XIE Yun-qing
,
YING Min-gang
2011, 18(2):126-132.
DOI: 10.3872/j.issn.1007-385X.2011.2.002
Abstract:
Objective:To isolate stem-like cells from hepatocellular carcinoma cell line PLC/PRF-5 and to study their miRNA profile. Methods：〖WTBZ〗ABCG2+ and ABCG2- PLC/PRF-5 cells were isolated from the PLC/PRF-5 cell line by magnetic activated cell sorting (MACS) method, and further identified by flow cytometry. The colony formation ability in soft agar and tumor formation ability in NOD/SCID mice of ABCG2+ and ABCG2- PLC/PRF-5 cells were observed. miRNA chip was adopted to screen the differentially expressed miRNAs between ABCG2+ and ABCG2- PLC/PRF-5 cells; and real-time PCR assay was used to confirm the results of miRNA chip. Results：〖WTBZ〗The purity of ABCG2+PLC/PRF-5 cells isolated by MACS method was (84.20±4.52)%. The colony number and size formed by ABCG2+ PLC/PRF-5 cells were more and larger than those formed by ABCG2- cells (47.17±10.50 vs 23.33±7.31, P<0.05). 1×104 ABCG2+ cells could form tumors with at least 5×105 cells needed for ABCG2- cells. The size of tumors generated by 5×105 ABCG2+ cells was larger than that by the ABCG2- cells (\[3.73±1.19\] cm3 vs \[0.72 ±0.57\] cm3, P<0.01). There are 20 miRNAs differentially expressed between ABCG2+ and ABCG2- cells, with 13 up-regulated and 7 down-regulated. Real-time PCR assay was applied to further verify the differential expression of hsa-miR-30a and hsa-miR-630, and the results were in accordance with those of miRNA chip. Conclusion:ABCG2+ PLC/PRF-5 cells in hepatocellular carcinoma cell line PLC/PRF-5 have the properties of cancer stem cells. Twenty miRNAs are differentially expressed between ABCG2+ and ABCG2- PLC/PRF-5 cells, which might play important roles in the carcinogenesis of hepatocellular carcinoma.
Objective:To isolate stem-like cells from hepatocellular carcinoma cell line PLC/PRF-5 and to study their miRNA profile. Methods：〖WTBZ〗ABCG2+ and ABCG2- PLC/PRF-5 cells were isolated from the PLC/PRF-5 cell line by magnetic activated cell sorting (MACS) method, and further identified by flow cytometry. The colony formation ability in soft agar and tumor formation ability in NOD/SCID mice of ABCG2+ and ABCG2- PLC/PRF-5 cells were observed. miRNA chip was adopted to screen the differentially expressed miRNAs between ABCG2+ and ABCG2- PLC/PRF-5 cells; and real-time PCR assay was used to confirm the results of miRNA chip. Results：〖WTBZ〗The purity of ABCG2+PLC/PRF-5 cells isolated by MACS method was (84.20±4.52)%. The colony number and size formed by ABCG2+ PLC/PRF-5 cells were more and larger than those formed by ABCG2- cells (47.17±10.50 vs 23.33±7.31, P<0.05). 1×104 ABCG2+ cells could form tumors with at least 5×105 cells needed for ABCG2- cells. The size of tumors generated by 5×105 ABCG2+ cells was larger than that by the ABCG2- cells (\[3.73±1.19\] cm3 vs \[0.72 ±0.57\] cm3, P<0.01). There are 20 miRNAs differentially expressed between ABCG2+ and ABCG2- cells, with 13 up-regulated and 7 down-regulated. Real-time PCR assay was applied to further verify the differential expression of hsa-miR-30a and hsa-miR-630, and the results were in accordance with those of miRNA chip. Conclusion:ABCG2+ PLC/PRF-5 cells in hepatocellular carcinoma cell line PLC/PRF-5 have the properties of cancer stem cells. Twenty miRNAs are differentially expressed between ABCG2+ and ABCG2- PLC/PRF-5 cells, which might play important roles in the carcinogenesis of hepatocellular carcinoma.
SUN Li-xin
,
RAN Yu-liang
,
YANG Wei-wei
,
SUN Li-chao
,
ZHANG Xiao-yan
,
RONG Yan
,
LIU Jun
,
YANG Zhi-hua
2011, 18(2):133-138.
DOI: 10.3872/j.issn.1007-385X.2011.2.003
Abstract:
Objective:To study the expression, subcellular localization and translocation of the targeted antigen of anti-lung cancer 15A1 mAb in lung cancer cells, and to study the inhibitory function of 15A1 mAb in vitro and in vivo against lung cancer cells, so as to provide target and antibody drug candidate for targeted therapy of lung cancer. Methods: The expression and localization of the targeted antigen of 15A1 mAb in lung cancer cell lines and lung cancer tissues were studied by immunofluorescence and flow cytometry. CCK-8 proliferation assay and tumor xenograft experiment were performed to evaluate the inhibitory function of 15A1 mAb aginst lung cancer cells in vitro and in vivo. Results:The antigen of 15A1 mAb was expressed in the cytoplasm and cell surface of 12 lung cancer cell lines, including human lung adenocarcinoma cell lines (A549, ANIP-793, GLC-82, Calu-3, H157, H1299), squamous carcinoma cell lines (GLC-P, H520), small cell lung cancer cell lines (H446, H209), and large cell lung cancer cell lines (PG, H460), with the highest expression level seen in the adenocarcinoma cell lines. The upregulated expression of the targeted antigen of 15A1 mAb was found in about 75% of human lung cancer tissues. Most targeted antigen translocated from the nucleus to cytoplasm in lung adenocarcinoma cells, but only part of the antigen translocated to the cytoplasm in lung squamous carcinoma cells. Some antigen also translocated to the cell membrane, which was more significantly in adenocarcinoma cells than in squamous carcinoma cells. The 15A1mAb greatly suppressed proliferation of lung cancer cells in vivo and in vitro, with the inhibitory rate being 25%〖KG-*2〗〖DK〗-80%. Conclusion: The 15A1 mAb can significantly inhibit growth of lung cancer cells in vivo and in vitro, and its targeted antigen is highly expressed on human lung cancer tissues and cells and can translocate to the cell surface, suggesting that 15A1 mAb might become a candidate target for lung cancer therapy.
Objective:To study the expression, subcellular localization and translocation of the targeted antigen of anti-lung cancer 15A1 mAb in lung cancer cells, and to study the inhibitory function of 15A1 mAb in vitro and in vivo against lung cancer cells, so as to provide target and antibody drug candidate for targeted therapy of lung cancer. Methods: The expression and localization of the targeted antigen of 15A1 mAb in lung cancer cell lines and lung cancer tissues were studied by immunofluorescence and flow cytometry. CCK-8 proliferation assay and tumor xenograft experiment were performed to evaluate the inhibitory function of 15A1 mAb aginst lung cancer cells in vitro and in vivo. Results:The antigen of 15A1 mAb was expressed in the cytoplasm and cell surface of 12 lung cancer cell lines, including human lung adenocarcinoma cell lines (A549, ANIP-793, GLC-82, Calu-3, H157, H1299), squamous carcinoma cell lines (GLC-P, H520), small cell lung cancer cell lines (H446, H209), and large cell lung cancer cell lines (PG, H460), with the highest expression level seen in the adenocarcinoma cell lines. The upregulated expression of the targeted antigen of 15A1 mAb was found in about 75% of human lung cancer tissues. Most targeted antigen translocated from the nucleus to cytoplasm in lung adenocarcinoma cells, but only part of the antigen translocated to the cytoplasm in lung squamous carcinoma cells. Some antigen also translocated to the cell membrane, which was more significantly in adenocarcinoma cells than in squamous carcinoma cells. The 15A1mAb greatly suppressed proliferation of lung cancer cells in vivo and in vitro, with the inhibitory rate being 25%〖KG-*2〗〖DK〗-80%. Conclusion: The 15A1 mAb can significantly inhibit growth of lung cancer cells in vivo and in vitro, and its targeted antigen is highly expressed on human lung cancer tissues and cells and can translocate to the cell surface, suggesting that 15A1 mAb might become a candidate target for lung cancer therapy.
2011, 18(2):139-143.
DOI: 10.3872/j.issn.1007-385X.2011.2.004
Abstract:
Objective:To study the relationship of CHFR CpG island methylation status on proliferation and apoptosis of esophageal cancer Eca109 cells. Methods: Eca109 cells were treated with different concentrations (2, 5, and 10 μmol/L) of 5-Aza-2′-deoxycytidine (5-Aza-CdR), and untreated Eca109 cells were used as control. Methylation-specific PCR (MSP) analysis was used to evaluate the CpG island methylation status of CHFR gene, and RT-PCR was used to detect the CHFR mRNA expression in Eca109 cells. MTT and flow cytometery were used to determine the proliferation and apoptosis of Eca109 cells treated with different concentrations of 5-Aza-CdR. Results: CHFR CpG island was hypermethylated in the untreated Eca109 cells, and methylated CHFR CpG was demethylated to different degrees in 5-Aza-CdR treatment groups. No expression of CHFR mRNA was found in untreated Eca109 cells, but the relative expression of CHFR mRNA was remarkably increased in 5-Aza-CdR (2, 5, and 10 μmol/L) treatment groups (0.174±0.010, 0.221±0.013, and 0.356±0014). Different concentrations of 5-Aza-CdR inhibited the proliferation (\[30.87±0.74\]%,\[44.60±0.79\]%, and \[56.67±035\]%), and promoted apoptosis of Eca109 cells (\[7.46±1.46\]%, \[16.27±1.61\]%, \[25.29±225\]% vs \[1.83±0.41\]%, P<001). Conclusion: 5-Aza-CdR can partly demethylate CHFR CpG island in esophageal cancer Eca109 cells, inducing CHFR mRNA expression, inhibiting proliferation and promoting apoptosis of Eca109 cells.
Objective:To study the relationship of CHFR CpG island methylation status on proliferation and apoptosis of esophageal cancer Eca109 cells. Methods: Eca109 cells were treated with different concentrations (2, 5, and 10 μmol/L) of 5-Aza-2′-deoxycytidine (5-Aza-CdR), and untreated Eca109 cells were used as control. Methylation-specific PCR (MSP) analysis was used to evaluate the CpG island methylation status of CHFR gene, and RT-PCR was used to detect the CHFR mRNA expression in Eca109 cells. MTT and flow cytometery were used to determine the proliferation and apoptosis of Eca109 cells treated with different concentrations of 5-Aza-CdR. Results: CHFR CpG island was hypermethylated in the untreated Eca109 cells, and methylated CHFR CpG was demethylated to different degrees in 5-Aza-CdR treatment groups. No expression of CHFR mRNA was found in untreated Eca109 cells, but the relative expression of CHFR mRNA was remarkably increased in 5-Aza-CdR (2, 5, and 10 μmol/L) treatment groups (0.174±0.010, 0.221±0.013, and 0.356±0014). Different concentrations of 5-Aza-CdR inhibited the proliferation (\[30.87±0.74\]%,\[44.60±0.79\]%, and \[56.67±035\]%), and promoted apoptosis of Eca109 cells (\[7.46±1.46\]%, \[16.27±1.61\]%, \[25.29±225\]% vs \[1.83±0.41\]%, P<001). Conclusion: 5-Aza-CdR can partly demethylate CHFR CpG island in esophageal cancer Eca109 cells, inducing CHFR mRNA expression, inhibiting proliferation and promoting apoptosis of Eca109 cells.
XIAO Qing
,
HUANG Chuan
,
FAN Xiao-hui
,
SONG De-zhi
,
LIANG Ying
,
GONG Jin-ling
,
WANG Li-fang
,
LIU Jin-ying
,
LAI Zhen-ping
2011, 18(2):144-148.
DOI: 10.3872/j.issn.1007-385X.2011.2.005
Abstract:
Objective:To investigate the effects of Newcastle disease virus 7793 strain (NDV 7793) on the growth of human colon carcinoma LoVo cell-transplanted tumors in nude mice and the possible mechanism. Methods: Mouse models of LoVo cell-transplanted tumor were established and were randomly divided into 3 groups: intravenously injected with PBS, 5-FU and NDV 7793 groups. Tumor growth was observed in different groups, the apoptosis and necrosis rates of tumor cells were detected by FCM, expressions of Bax and Bcl-2 proteins were analyzed by immunohistochemical method, cyto-C level in tumor tissues was detected by cyto-C kit, and the concentration of TNF-α in tumor tissues was examined by ELISA. Results: NDV 7793 significantly inhibited the growth of LoVo-transplanted tumors compared with 5-FU (50.14% vs 3714%, P<0.05). The apoptosis rate of LoVo-transplanted tumor cells in NDV 7793 group was significantly higher than that in 5-FU group (\[28.7±1.5\]% vs \[1.46±0.3\]%), and LoVo-transplanted tumor cells had a similar apoptosis rate and necrosis rate in NDV 7793 group (\[28.7±1.5\]% vs \[27.80±3.32\]%). NDV 7793 enhanced the expression of Bax, but not Bcl-2, in LoVo-transplanted tumor tissues, NDV 7793 also increased the cyto-C (\[2.28±0.68\] vs \[0.68±0.13\] μg/μl) and TNF-α levels (\[489.6±5.2\] vs \[167.9±3.9\] pg/ml) in LoVo-transplanted tumor tissues. Conclusion：NDV 7793 can inhibit the growth of human colon carcinoma LoVo cell-transplanted tumors, which may be related to the up-regulation of Bax, cyto-C and TNF-α and the subsequent apoptosis of tumor cells.
Objective:To investigate the effects of Newcastle disease virus 7793 strain (NDV 7793) on the growth of human colon carcinoma LoVo cell-transplanted tumors in nude mice and the possible mechanism. Methods: Mouse models of LoVo cell-transplanted tumor were established and were randomly divided into 3 groups: intravenously injected with PBS, 5-FU and NDV 7793 groups. Tumor growth was observed in different groups, the apoptosis and necrosis rates of tumor cells were detected by FCM, expressions of Bax and Bcl-2 proteins were analyzed by immunohistochemical method, cyto-C level in tumor tissues was detected by cyto-C kit, and the concentration of TNF-α in tumor tissues was examined by ELISA. Results: NDV 7793 significantly inhibited the growth of LoVo-transplanted tumors compared with 5-FU (50.14% vs 3714%, P<0.05). The apoptosis rate of LoVo-transplanted tumor cells in NDV 7793 group was significantly higher than that in 5-FU group (\[28.7±1.5\]% vs \[1.46±0.3\]%), and LoVo-transplanted tumor cells had a similar apoptosis rate and necrosis rate in NDV 7793 group (\[28.7±1.5\]% vs \[27.80±3.32\]%). NDV 7793 enhanced the expression of Bax, but not Bcl-2, in LoVo-transplanted tumor tissues, NDV 7793 also increased the cyto-C (\[2.28±0.68\] vs \[0.68±0.13\] μg/μl) and TNF-α levels (\[489.6±5.2\] vs \[167.9±3.9\] pg/ml) in LoVo-transplanted tumor tissues. Conclusion：NDV 7793 can inhibit the growth of human colon carcinoma LoVo cell-transplanted tumors, which may be related to the up-regulation of Bax, cyto-C and TNF-α and the subsequent apoptosis of tumor cells.
2011, 18(2):155-159.
DOI: 10.3872/j.issn.1007-385X.2011.2.007
Abstract:
Objective:To explore the role of artesunate (Art) in resistance-reversal of Eca109/ADM cells and the related mechanism. Methods：The present study was divided into 4 groups: normal saline (NS) control group, Art (0.1 μmol/L) group, ADM (0.2 μg/ml) group, and Art (0.1 μmol/L)+ADM (0.2 μg/ml) group. The apoptosis rate, ADM content and ABCG2 (ATP-binding cassette transporter G2) protein expression in Eca109/ADM cells were detected by flow cytometry 48 h after treated with Art, ADM, and Art+ADM. ABCG2 protein expression in Eca109/ADM cells was further examined by Western blotting analysis. Results：The apoptosis rate of Eca109/ADM cells in Art+ADM group was (12.89±0.87)%, being significantly higher than that in the Art group (1.58±0.12)%, ADM group (6.55±090)% and NS group (1.44±0.10)% (P<0.05). Art increased the sensitivity of Eca109/ADM cells to ADM. Flow cytometry results showed that the ABCG2 protein expression in Eca109/ADM cells of Art+ADM group (644.60±3.21) was significantly lower than that in ADM group (659.15±4.59) and NS group (658.14±6.88) (P<0.05), but was similar to that in Art group (644.31±3.96) (P>0.05). Western blotting analysis results were consistent with those detected by flow cytometry. The ABCG2 protein expression of Eca109/ADM cells in Art group (0.70±0.02) was significantly decreased compared with NS group (0.80±0.03) (P<0.05), and that in Art+ADM group (0.71±0.04) was also decreased compared with ADM group (0.81±0.05) (P<0.05). The ADM content of Eca109/ADM cells in Art+ADM group was significantly higher than those in Art, ADM and NS groups (848.02±5.04 vs 763.29±4.02, 800.25±3.84, 763.88±2.03; P<0.01). Conclusion: Art can decrease ABCG2 protein expression and increase ADM content in Eca109/ADM cells, and it can also reverse the drug resistance of Eca109/ADM cells.
Objective:To explore the role of artesunate (Art) in resistance-reversal of Eca109/ADM cells and the related mechanism. Methods：The present study was divided into 4 groups: normal saline (NS) control group, Art (0.1 μmol/L) group, ADM (0.2 μg/ml) group, and Art (0.1 μmol/L)+ADM (0.2 μg/ml) group. The apoptosis rate, ADM content and ABCG2 (ATP-binding cassette transporter G2) protein expression in Eca109/ADM cells were detected by flow cytometry 48 h after treated with Art, ADM, and Art+ADM. ABCG2 protein expression in Eca109/ADM cells was further examined by Western blotting analysis. Results：The apoptosis rate of Eca109/ADM cells in Art+ADM group was (12.89±0.87)%, being significantly higher than that in the Art group (1.58±0.12)%, ADM group (6.55±090)% and NS group (1.44±0.10)% (P<0.05). Art increased the sensitivity of Eca109/ADM cells to ADM. Flow cytometry results showed that the ABCG2 protein expression in Eca109/ADM cells of Art+ADM group (644.60±3.21) was significantly lower than that in ADM group (659.15±4.59) and NS group (658.14±6.88) (P<0.05), but was similar to that in Art group (644.31±3.96) (P>0.05). Western blotting analysis results were consistent with those detected by flow cytometry. The ABCG2 protein expression of Eca109/ADM cells in Art group (0.70±0.02) was significantly decreased compared with NS group (0.80±0.03) (P<0.05), and that in Art+ADM group (0.71±0.04) was also decreased compared with ADM group (0.81±0.05) (P<0.05). The ADM content of Eca109/ADM cells in Art+ADM group was significantly higher than those in Art, ADM and NS groups (848.02±5.04 vs 763.29±4.02, 800.25±3.84, 763.88±2.03; P<0.01). Conclusion: Art can decrease ABCG2 protein expression and increase ADM content in Eca109/ADM cells, and it can also reverse the drug resistance of Eca109/ADM cells.
SUN Mao-ben
,
GUO Kun-yuan
,
HU Liang-shan
,
DENG Lan
,
LI Yu-hua
,
SONG Chao-yang
,
HE Yan-jie
,
CHEN Jun
2011, 18(2):160-164.
DOI: 10.3872/j.issn.1007-385X.2011.2.008
Abstract:
Objective:To investigate the apoptosis-inducing effect of honokiol (HNK) on human acute myeloid leukemia KG1a cells and the possible mechanism. Methods: KG1a cells were treated with different concentrations of HNK, and then the proliferation of KG1a cells was detected by XTT assay. Flow cytometry was performed to examine the cell cycle and apoptosis of KG1a cells after HNK (0, 5, 10 μg/ml) treatment. RT-PCR technique was used to detect the expression of apoptosis-related genes (〖STBX〗Bcl-2, Bid, Bax, Bak, Bad, P53 and NF-κB〖STBZ〗) in KG1a cells. Results: HNK (2.5, 5, 8, 10, 15, 20, and 40 μg/ml) significantly inhibited the growth of KG1a cells in a time- and dose-dependent manner (P<0.01), and IC50 concentrations in 24 h, 48 h were 10.23 μg/ml and 8.25 μg/ml, respectively. Flow cytometry results revealed that KG1a cells were arrested at G0/G1 phase after treated with HNK; the early apoptotic rates of KG1a cells after HNK treatments (5 μg/ml and 10 μg/ml) were significantly higher than those in control group (0 μg/ml), with apoptotic rates being (\[11.16±1.27\]%, \[21.46±3.13\]% vs \[6.03±1.10\]%, P<0.01). Meanwhile, RT-PCR revealed the mRNA expression of apoptosis-promoting gene Bax was significantly increased, with Bad slightly improved after HNK treatments compared with control group; meantime, the apoptosis-inhibiting gene (NF-κB) was markedly decreased. Conclusion: HNK can induce apoptosis of KG1a cells, which might be related to increased expression of Bax, Bad genes, and decreased expression of NF-κB gene.
Objective:To investigate the apoptosis-inducing effect of honokiol (HNK) on human acute myeloid leukemia KG1a cells and the possible mechanism. Methods: KG1a cells were treated with different concentrations of HNK, and then the proliferation of KG1a cells was detected by XTT assay. Flow cytometry was performed to examine the cell cycle and apoptosis of KG1a cells after HNK (0, 5, 10 μg/ml) treatment. RT-PCR technique was used to detect the expression of apoptosis-related genes (〖STBX〗Bcl-2, Bid, Bax, Bak, Bad, P53 and NF-κB〖STBZ〗) in KG1a cells. Results: HNK (2.5, 5, 8, 10, 15, 20, and 40 μg/ml) significantly inhibited the growth of KG1a cells in a time- and dose-dependent manner (P<0.01), and IC50 concentrations in 24 h, 48 h were 10.23 μg/ml and 8.25 μg/ml, respectively. Flow cytometry results revealed that KG1a cells were arrested at G0/G1 phase after treated with HNK; the early apoptotic rates of KG1a cells after HNK treatments (5 μg/ml and 10 μg/ml) were significantly higher than those in control group (0 μg/ml), with apoptotic rates being (\[11.16±1.27\]%, \[21.46±3.13\]% vs \[6.03±1.10\]%, P<0.01). Meanwhile, RT-PCR revealed the mRNA expression of apoptosis-promoting gene Bax was significantly increased, with Bad slightly improved after HNK treatments compared with control group; meantime, the apoptosis-inhibiting gene (NF-κB) was markedly decreased. Conclusion: HNK can induce apoptosis of KG1a cells, which might be related to increased expression of Bax, Bad genes, and decreased expression of NF-κB gene.
YANG De-meng
,
WANG Jie
,
PENG Yuan-yuan
,
WANG Li
,
XIA Bing
,
ZHANG Hong-bin
,
YANG Chuang-hong
,
ZHANG Hai-yan
2011, 18(2):165-169.
DOI: 10.3872/j.issn.1007-385X.2011.2.009
Abstract:
Objective:To investigate the effect of bone sialoprotein (BSP) on the PI3K-AKT signaling pathway in breast cancer MDA-MB-231 cells. Methods: BSP-silenced MDA-MB-231 cells were treated with recombinant human BSP (rhBSP) and the PI3K-AKT specific inhibitor LY294002. Western blotting analysis was used to detect the phosphorylation of AKT, qPCR was conducted to evaluate caspase-3, cyclin D1 mRNA expressions, and the proliferation of cells was analyzed by MTT assay. Results: Compared with the 231BO-Scrambled cells in control group, BSP protein expression in BSP-silenced 231BO-BSP27 cells was significantly lower (74.32±2.18)% (P<0.01), and expression level of AKT phosphorylation was also significantly lower (33.30±2.61) % (P<0.01), resulting in up-regulation of caspase-3 mRNA level (1.000±0.000 vs 1.733±0.039, P<0.01) , down-regulation of cyclin D1 mRNA (1.000±0.000 vs 0370±0.012, P<0. 01),and the inhibition of 231BO-BSP27 cells growth (P<0.05). After treatment with exogenous rhBSP, the phosphorylation of AKT was increased in both 231BO-Scrambled and 231BO-BSP27 cells \[(17.86±2.27)%, (33.78±1.51)%, P<0.01\]. rhBSP treatment decreased caspase-3 mRNA (1.000±0.039 vs 0.541±0.091, P<0.01) , and increased cyclin D1 mRNA (1.000±0.000 vs 2.921±0.032, P<0.01) expression in 231BO-BSP27 cells, and stimulated the proliferation of 231BO-Scrambled and 231BO-BSP27 cells (P<0.01). Furthermore, rhBSP-induced activation of AKT was reversed by LY294002 in 231BO-Scrambled and 231BO-BSP27 cells (P<0.05), with an increase in caspase-3 mRNA and decrease in cyclin D1 mRNA expression in 231BO-BSP27 cells (all P<0.01), causing proliferation inhibition in 231BO-Scrambled and 231BO-BSP27 cells (P<0.01). Conclusion: BSP can regulate the mRNA expressions of caspase-3 and cyclin D1, and affect the proliferation of breast cancer MDA-MB-231 cells through the PI3K-AKT signaling pathway.
Objective:To investigate the effect of bone sialoprotein (BSP) on the PI3K-AKT signaling pathway in breast cancer MDA-MB-231 cells. Methods: BSP-silenced MDA-MB-231 cells were treated with recombinant human BSP (rhBSP) and the PI3K-AKT specific inhibitor LY294002. Western blotting analysis was used to detect the phosphorylation of AKT, qPCR was conducted to evaluate caspase-3, cyclin D1 mRNA expressions, and the proliferation of cells was analyzed by MTT assay. Results: Compared with the 231BO-Scrambled cells in control group, BSP protein expression in BSP-silenced 231BO-BSP27 cells was significantly lower (74.32±2.18)% (P<0.01), and expression level of AKT phosphorylation was also significantly lower (33.30±2.61) % (P<0.01), resulting in up-regulation of caspase-3 mRNA level (1.000±0.000 vs 1.733±0.039, P<0.01) , down-regulation of cyclin D1 mRNA (1.000±0.000 vs 0370±0.012, P<0. 01),and the inhibition of 231BO-BSP27 cells growth (P<0.05). After treatment with exogenous rhBSP, the phosphorylation of AKT was increased in both 231BO-Scrambled and 231BO-BSP27 cells \[(17.86±2.27)%, (33.78±1.51)%, P<0.01\]. rhBSP treatment decreased caspase-3 mRNA (1.000±0.039 vs 0.541±0.091, P<0.01) , and increased cyclin D1 mRNA (1.000±0.000 vs 2.921±0.032, P<0.01) expression in 231BO-BSP27 cells, and stimulated the proliferation of 231BO-Scrambled and 231BO-BSP27 cells (P<0.01). Furthermore, rhBSP-induced activation of AKT was reversed by LY294002 in 231BO-Scrambled and 231BO-BSP27 cells (P<0.05), with an increase in caspase-3 mRNA and decrease in cyclin D1 mRNA expression in 231BO-BSP27 cells (all P<0.01), causing proliferation inhibition in 231BO-Scrambled and 231BO-BSP27 cells (P<0.01). Conclusion: BSP can regulate the mRNA expressions of caspase-3 and cyclin D1, and affect the proliferation of breast cancer MDA-MB-231 cells through the PI3K-AKT signaling pathway.
9 Cisplatin-resistance increases proliferation, invasion and migration of ovarian cancer OVCAR-3 cells
2011, 18(2):170-174.
DOI: 10.3872/j.issn.1007-385X.2011.2.010
Abstract:
Objective:To investigate the differences in biological behavior between cisplatin-sensitive (OVCAR-3 cells) and cisplatin-resistant human ovarian cancer cells (OVCAR-3/CDDP cells), and to explore the possible mechanisms. Methods: OVCAR-3 and OVCAR-3/CDDP cells in logarithmic phase were collected, their proliferation were assessed by soft agar colony formation assay, their in vitro and in vivo invasion and migration abilities were measured by Transwell chamber assay, anoikis assay and subcutaneous transplanted-tumor formation assay in nude mice, and the expressions of MMP-2 and MMP-9 in transplanted tumor tissues were examined by immunohistochemical assay. Results: Colony formation rate of OVCAR-3/CDDP cells was significantly increased compared with OVCAR-3 cells (\[0.66%±009%\] vs \[0.31%±0.07%\], P<0.05). Compared with OVCAR-3 cells, OVCAR-3/CDDP cells had significantly higher migration capability (\[233.1±8.5\] vs \[167.4±5.9\], P<0.01) and invasive capability (\[143.6±9.1\] vs \[95.8±6.2\], P<0.01). OVCAR-3/CDDP cells tended to aggregate and their apoptosis index was decreased compared with that OVCAR-3 cells (\[7.78±1.32\]% vs \[15.41±1.26\]%, P<0.01). The expressions of MMP-2 and MMP-9 in transplanted tumor tissues of OVCAR-3/CDDP cell group were up-regulated compared with those of OVCAR-3 cell group (P<0.05). Conclusion: The proliferation, anti-anoikis, invasion and migration abilities of cisplatin-resistant OVCAR-3/CDDP cells are greatly increased, which might be related with the up-regulation of MMP-2 and MMP-9.
Objective:To investigate the differences in biological behavior between cisplatin-sensitive (OVCAR-3 cells) and cisplatin-resistant human ovarian cancer cells (OVCAR-3/CDDP cells), and to explore the possible mechanisms. Methods: OVCAR-3 and OVCAR-3/CDDP cells in logarithmic phase were collected, their proliferation were assessed by soft agar colony formation assay, their in vitro and in vivo invasion and migration abilities were measured by Transwell chamber assay, anoikis assay and subcutaneous transplanted-tumor formation assay in nude mice, and the expressions of MMP-2 and MMP-9 in transplanted tumor tissues were examined by immunohistochemical assay. Results: Colony formation rate of OVCAR-3/CDDP cells was significantly increased compared with OVCAR-3 cells (\[0.66%±009%\] vs \[0.31%±0.07%\], P<0.05). Compared with OVCAR-3 cells, OVCAR-3/CDDP cells had significantly higher migration capability (\[233.1±8.5\] vs \[167.4±5.9\], P<0.01) and invasive capability (\[143.6±9.1\] vs \[95.8±6.2\], P<0.01). OVCAR-3/CDDP cells tended to aggregate and their apoptosis index was decreased compared with that OVCAR-3 cells (\[7.78±1.32\]% vs \[15.41±1.26\]%, P<0.01). The expressions of MMP-2 and MMP-9 in transplanted tumor tissues of OVCAR-3/CDDP cell group were up-regulated compared with those of OVCAR-3 cell group (P<0.05). Conclusion: The proliferation, anti-anoikis, invasion and migration abilities of cisplatin-resistant OVCAR-3/CDDP cells are greatly increased, which might be related with the up-regulation of MMP-2 and MMP-9.
2011, 18(2):175-180.
DOI: 10.3872/j.issn.1007-385X.2011.2.011
Abstract:
Objective:To investigate the inhibitory effect of basic fibroblast growth factor monoclonal antibody (bFGF-mAb) combined with radiotherapy against B16-transplanted tumors in mice. Methods: bFGF-mAb was prepared and purified. B16-transplanted melanoma tumor models were established and the mice were randomly divided into 4 groups: control group, radiotherapy group, bFGF-mAb group, and bFGF-mAb combined with radiotherapy group. Tumor volumes were measured in different treatment groups. Twenty days after treatment, the tumors were collected and weighted, and the inhibitory rates of tumor growth were calculated. TUNEL staining was used to detect the apoptosis rate of transplanted tumors; immunohistochemical method was used to examine the positive expression of bFGF, vascular endothelial growth factor (VEGF) and microvessel density (MVD) in transplanted tumor tissues. Results: The inhibitory rate of tumor growth in the combined treatment group was significantly higher than those in radiotherapy group and bFGF-mAb treatment group (48.76% vs 12.17%, 30.49%, P<0.05). The radiotherapy sensitization enhancement ratio of the combined treatment group was 2.37 times that in the radiotherapy group. The apoptosis rate of transplanted tumor cells in the combined treatment group was significantly increased compared with those in the control, radiotherapy, bFGF-mAb treatment groups (\[58.56±6.47\]% vs \[17.21±2.86\]%, \[28.45±5.47\]%, \[10.62±1.73\]%; P<0.05 or P<001), with bFGF, VEGF expression and MVD being significantly decreased (P<005). Conclusion: bFGF-mAb combined with radiotherapy have synergistic inhibitory effect on the growth of B16-transplanted melanoma tumors, and it can increase the radio-sensitivity of tumor cells by reducing the expressions of bFGF and VEGF, decreasing angiogenesis, and promoting apoptosis.
Objective:To investigate the inhibitory effect of basic fibroblast growth factor monoclonal antibody (bFGF-mAb) combined with radiotherapy against B16-transplanted tumors in mice. Methods: bFGF-mAb was prepared and purified. B16-transplanted melanoma tumor models were established and the mice were randomly divided into 4 groups: control group, radiotherapy group, bFGF-mAb group, and bFGF-mAb combined with radiotherapy group. Tumor volumes were measured in different treatment groups. Twenty days after treatment, the tumors were collected and weighted, and the inhibitory rates of tumor growth were calculated. TUNEL staining was used to detect the apoptosis rate of transplanted tumors; immunohistochemical method was used to examine the positive expression of bFGF, vascular endothelial growth factor (VEGF) and microvessel density (MVD) in transplanted tumor tissues. Results: The inhibitory rate of tumor growth in the combined treatment group was significantly higher than those in radiotherapy group and bFGF-mAb treatment group (48.76% vs 12.17%, 30.49%, P<0.05). The radiotherapy sensitization enhancement ratio of the combined treatment group was 2.37 times that in the radiotherapy group. The apoptosis rate of transplanted tumor cells in the combined treatment group was significantly increased compared with those in the control, radiotherapy, bFGF-mAb treatment groups (\[58.56±6.47\]% vs \[17.21±2.86\]%, \[28.45±5.47\]%, \[10.62±1.73\]%; P<0.05 or P<001), with bFGF, VEGF expression and MVD being significantly decreased (P<005). Conclusion: bFGF-mAb combined with radiotherapy have synergistic inhibitory effect on the growth of B16-transplanted melanoma tumors, and it can increase the radio-sensitivity of tumor cells by reducing the expressions of bFGF and VEGF, decreasing angiogenesis, and promoting apoptosis.
2011, 18(2):181-185.
DOI: 10.3872/j.issn.1007-385X.2011.2.012
Abstract:
Objective:To study the in vitro and in vivo anti-tumor activities of Fomitopsis pinicola extract and their possible mechanisms. Methods: Fomitopsis pinicola was extracted by alcohol and the resulting acid ethyl esters extract was named FP-I. The inhibitory effects of FP-I on proliferation of mouse hepatocellular carcinoma H22 cells and mouse sarcoma S180 cells were detected by MTT assay in vitro. S180-implanted mouse model was established, and the tumor weight, tumor inhibitory rate, thymus index, spleen index, peripheral leukocyte number, peripheral lymphocyte rate, and associate tumor erythocyte rosette rate (ATER) were examined in different group. Effect of FP-I on apoptosis of S180-implanted tumor cells and the histological changes of the heart, liver, kidney, thymus and spleen were observed by H-E staining. Results: The inhibitory rates of FP-I (50, 100, 200, 400 μg/ml) on proliferation of S180 cells were 22.35%, 32.49%, 40.01%, and 74.01%, respectively; and those on H22 cells were 45.19%, 51.10%, 66.37%, and 82.40%, rspectively. The inhibitory rates of FP-I (25, 50, 100 μg/ml) on growth of S180-implanted tumors were 79.92%, 66.18% and 78.45%, respectively, and that of CTX (30 mg/kg) positive control was 84.10%. Proportion of lymphocyte in periphoral blood, thymus index and ATER of S180-bearing mice were significantly increased in both the high-and medium-dose FP-I groups (P<0.05 or P<0.01). S180-implanted tumor cells in all FP-I treatment groups showed typical apoptotic morphology. Conclusion: Fomitopsis pinicola extract has anti-tumor activity, which might relate to its ability to improve the immune function of mice and to induce apoptosis of tumor cells.
Objective:To study the in vitro and in vivo anti-tumor activities of Fomitopsis pinicola extract and their possible mechanisms. Methods: Fomitopsis pinicola was extracted by alcohol and the resulting acid ethyl esters extract was named FP-I. The inhibitory effects of FP-I on proliferation of mouse hepatocellular carcinoma H22 cells and mouse sarcoma S180 cells were detected by MTT assay in vitro. S180-implanted mouse model was established, and the tumor weight, tumor inhibitory rate, thymus index, spleen index, peripheral leukocyte number, peripheral lymphocyte rate, and associate tumor erythocyte rosette rate (ATER) were examined in different group. Effect of FP-I on apoptosis of S180-implanted tumor cells and the histological changes of the heart, liver, kidney, thymus and spleen were observed by H-E staining. Results: The inhibitory rates of FP-I (50, 100, 200, 400 μg/ml) on proliferation of S180 cells were 22.35%, 32.49%, 40.01%, and 74.01%, respectively; and those on H22 cells were 45.19%, 51.10%, 66.37%, and 82.40%, rspectively. The inhibitory rates of FP-I (25, 50, 100 μg/ml) on growth of S180-implanted tumors were 79.92%, 66.18% and 78.45%, respectively, and that of CTX (30 mg/kg) positive control was 84.10%. Proportion of lymphocyte in periphoral blood, thymus index and ATER of S180-bearing mice were significantly increased in both the high-and medium-dose FP-I groups (P<0.05 or P<0.01). S180-implanted tumor cells in all FP-I treatment groups showed typical apoptotic morphology. Conclusion: Fomitopsis pinicola extract has anti-tumor activity, which might relate to its ability to improve the immune function of mice and to induce apoptosis of tumor cells.
2011, 18(2):186-189.
DOI: 10.3872/j.issn.1007-385X.2011.2.013
Abstract:
Objective:To explore the expression of apoptotic inhibitory protein survivin in pancreatic cancer cell line PaTu8988, and to study its role in the drug resistance of PaTu8988 cells to gemcitabine (GEM). Methods: The inhibitory effect of GEM (0.01, 0.1, 1.0, 2.5, 5.0, and 10.0 μg/ml) on PaTu8988 cells was detected by MTT assay; apoptosis rate of PaTu8988 cells treated with GEM was determined by flow cytometry; and the survivin mRNA expression in PaTu8988 cells was examined by RT-PCR. Results: High dosage of GEM (≥1.0 μg/ml) greatly inhibited growth and promoted apoptosis of PaTu8988 cells, while low dosage of GEM (0.01, 0.1 μg/ml) showed no effects. Low dose of GEM time-dependently increased expression of survinin mRNA in PaTu8988 cells; high dosage of GEM gradually inhibited survivin mRNA expression within the first 48 h, and then survivin mRNA expression gradually increased as time went by. Conclusion: Survivin mRNA is highly expressed in pancreatic cancer cell line PaTu8988, which may be one of the reasons for drug resistance to GEM.
Objective:To explore the expression of apoptotic inhibitory protein survivin in pancreatic cancer cell line PaTu8988, and to study its role in the drug resistance of PaTu8988 cells to gemcitabine (GEM). Methods: The inhibitory effect of GEM (0.01, 0.1, 1.0, 2.5, 5.0, and 10.0 μg/ml) on PaTu8988 cells was detected by MTT assay; apoptosis rate of PaTu8988 cells treated with GEM was determined by flow cytometry; and the survivin mRNA expression in PaTu8988 cells was examined by RT-PCR. Results: High dosage of GEM (≥1.0 μg/ml) greatly inhibited growth and promoted apoptosis of PaTu8988 cells, while low dosage of GEM (0.01, 0.1 μg/ml) showed no effects. Low dose of GEM time-dependently increased expression of survinin mRNA in PaTu8988 cells; high dosage of GEM gradually inhibited survivin mRNA expression within the first 48 h, and then survivin mRNA expression gradually increased as time went by. Conclusion: Survivin mRNA is highly expressed in pancreatic cancer cell line PaTu8988, which may be one of the reasons for drug resistance to GEM.
2011, 18(2):190-195.
DOI: 10.3872/j.issn.1007-385X.2011.2.014
Abstract:
Objective:To compare the microRNA expression profiles between gastrointestinal stromal tumors (GISTs) and extra-gastrointestinal stromal tumors (EGISTs), and to evaluate their relationship with primary sites and mutant status of tumors. Methods: Formalin-fixed paraffin-embedded tissues of 5 GISTs and 3 EGISTs were evaluated for differential microRNA (miRNA) expression signatures using Agilent microarrays containing 866 human miRNAs and mutation status of these samples were analyzed by PCR amplification and direct sequencing. Results: Unsupervised hierarchical clustering analysis revealed that 8 samples had been divided into 3 clusters. One gastric GIST and 1 EGIST with the same mutation grouped together and the remaining 2 gastric GISTs also formed a cluster. The other 4 samples including 2 intestinal GISTs and 2 EGISTs could be divided into a third separate cluster. And then we divided 5 GSITs samples into 2 groups according to their anatomical location (stomach and intestine) and assigned 3 EGISTs samples to the group with a similar miRNA expression pattern. Twelve miRNAs, most of which were predicted to participate in the regulation of KIT/PDGFRA signaling, were found to be differentially expressed between the two groups; among them, 5 miRNAs known to be involved in tumor progression were found down-regulated in intestinal group. In addition, only 3 miRNAs were found differentially expressed between GISTs and EGISTs. Conclusion:GISTs and EGISTs have similar miRNA expression patterns related with sites and mutation status; miRNA expression patterns may help to differentiate GISTs at the molecular level.
Objective:To compare the microRNA expression profiles between gastrointestinal stromal tumors (GISTs) and extra-gastrointestinal stromal tumors (EGISTs), and to evaluate their relationship with primary sites and mutant status of tumors. Methods: Formalin-fixed paraffin-embedded tissues of 5 GISTs and 3 EGISTs were evaluated for differential microRNA (miRNA) expression signatures using Agilent microarrays containing 866 human miRNAs and mutation status of these samples were analyzed by PCR amplification and direct sequencing. Results: Unsupervised hierarchical clustering analysis revealed that 8 samples had been divided into 3 clusters. One gastric GIST and 1 EGIST with the same mutation grouped together and the remaining 2 gastric GISTs also formed a cluster. The other 4 samples including 2 intestinal GISTs and 2 EGISTs could be divided into a third separate cluster. And then we divided 5 GSITs samples into 2 groups according to their anatomical location (stomach and intestine) and assigned 3 EGISTs samples to the group with a similar miRNA expression pattern. Twelve miRNAs, most of which were predicted to participate in the regulation of KIT/PDGFRA signaling, were found to be differentially expressed between the two groups; among them, 5 miRNAs known to be involved in tumor progression were found down-regulated in intestinal group. In addition, only 3 miRNAs were found differentially expressed between GISTs and EGISTs. Conclusion:GISTs and EGISTs have similar miRNA expression patterns related with sites and mutation status; miRNA expression patterns may help to differentiate GISTs at the molecular level.
2011, 18(2):196-200.
DOI: 10.3872/j.issn.1007-385X.2011.2.015
Abstract:
Objective:To retrospective study the clinical characteristics of melanoma patients receiving adjuvant therapy after surgery and to evaluate their disease-free survival (DFS) after different treatment options. Methods: Four hundred and fifty patients with malignant melanoma (stageⅠ to Ⅲ), who had received surgery in our center from January 2006 to July 2009, were included in the present study. The patients were from 28 cities, including 239 males and 211 females, with an age range of 12〖KG-*2〗〖DK〗-85 years. After surgery, the patients received different adjuvant therapies, including high-dose interferon treatment (interferon 22 000 000 IU iv. 5 times a week for 4 weeks, or 9 000 000 IU sc. 3 times a week for 11 months), chemotherapy (most cases were treated with dacarbazine or temozolomide; some were treated with cisplatin, paclitaxel and vincristine combination), radiochemotherapy, or radiotherapy (primary tumors or tumor-draining lymph nodes, 40〖KG-*2〗〖DK〗-60 Gy). Results: The surgical options included local resection of the primary lesion, expanded resection of the primary lesion, and extended resection and removal of regional lymph node. The median DFS were 15 months, 24 months and 13 months, respectively. Totally 184 patients did not receive any treatment after surgery, 84 received chemotherapy, 25 received radiochemotherapy, and 2 received radiotherapy; their median DFS were 13 months, 20 months, 29 months and 23 months, respectively. Totally 155 patients received high-dose interferon treatment and the median DFS was not reached. The side effects of chemotherapy included gastrointestinal side effects, bone marrow suppression and liver damage. The side effects of high-dose interferon included leukopenia, fever, fatigue, increased aminotransferase and anorexia. The incidences of severe leukopenia and increased aminotransferase (reaching 3 or 4 degree) were 15% and 10%, respectively. All patients returned to normal after symptomatic treatment. Conclusion:Surgical resection and adjuvant therapy strategy are extremely important for DFS of patients with malignant melanoma in stageⅠ to Ⅲ. The best DFS can be obtained in malignant melanoma patients who receive high-dose interferon therapy after surgery, with better safety.
Objective:To retrospective study the clinical characteristics of melanoma patients receiving adjuvant therapy after surgery and to evaluate their disease-free survival (DFS) after different treatment options. Methods: Four hundred and fifty patients with malignant melanoma (stageⅠ to Ⅲ), who had received surgery in our center from January 2006 to July 2009, were included in the present study. The patients were from 28 cities, including 239 males and 211 females, with an age range of 12〖KG-*2〗〖DK〗-85 years. After surgery, the patients received different adjuvant therapies, including high-dose interferon treatment (interferon 22 000 000 IU iv. 5 times a week for 4 weeks, or 9 000 000 IU sc. 3 times a week for 11 months), chemotherapy (most cases were treated with dacarbazine or temozolomide; some were treated with cisplatin, paclitaxel and vincristine combination), radiochemotherapy, or radiotherapy (primary tumors or tumor-draining lymph nodes, 40〖KG-*2〗〖DK〗-60 Gy). Results: The surgical options included local resection of the primary lesion, expanded resection of the primary lesion, and extended resection and removal of regional lymph node. The median DFS were 15 months, 24 months and 13 months, respectively. Totally 184 patients did not receive any treatment after surgery, 84 received chemotherapy, 25 received radiochemotherapy, and 2 received radiotherapy; their median DFS were 13 months, 20 months, 29 months and 23 months, respectively. Totally 155 patients received high-dose interferon treatment and the median DFS was not reached. The side effects of chemotherapy included gastrointestinal side effects, bone marrow suppression and liver damage. The side effects of high-dose interferon included leukopenia, fever, fatigue, increased aminotransferase and anorexia. The incidences of severe leukopenia and increased aminotransferase (reaching 3 or 4 degree) were 15% and 10%, respectively. All patients returned to normal after symptomatic treatment. Conclusion:Surgical resection and adjuvant therapy strategy are extremely important for DFS of patients with malignant melanoma in stageⅠ to Ⅲ. The best DFS can be obtained in malignant melanoma patients who receive high-dose interferon therapy after surgery, with better safety.
2011, 18(2):201-205.
DOI: 10.3872/j.issn.1007-385X.2011.2.016
Abstract:
Objective:To investigate the correlation of Toll-like receptors（TLR）4 and TLR9 expression with the pathological stages and metastases of gastric cancer. Methods: Forty-two gastric cancer and paracancerous tissues were included in the present study (patients were diagnosed in No.174 Hospital of PLA from Jan.1 2009 to Oct. 2009). The mRNA and protein expressions of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 in the gastric cancer and paracancerous tissues were detected by RT-PCR and Western blotting analysis, respectively. Results: The expressions of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 mRNA were significantly higher in the gastric cancer tissues than those in the paracancerous tissues (1.29±0.03 vs 0.53±0.02, 0.99±0.04 vs 0.22±0.05；P<0.05). T3+T4 gastric cancer tissues demonstrated significantly higher levels of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 mRNA than the T1+T2 tissues (P<0.05). Significantly higher levels of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 mRNA were observed in patients with distant metastases than in those without (P<0.05). However, no correlation was observed between 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 mRNA expression with the age, gender or lymphatic metastasis status of patients. The expressions of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 protein were significantly higher in the gastric cancer tissues than those in the paracancerous tissues (1.36±0.05 vs 0.48±0.04, 112±0.05 vs 0.34±0.03;P<0.05), T3+T4 gastric cancer tissues demonstrated significantly higher levels of TLR4 and TLR9 proteins than the T1+T2 tissues (P<0.05). Conclusions: TLR4 and TLR9 are up-regulated in gastric carcinoma tissues, and TLR4/9 expression is correlated with the TNM staging and distant metastasis. These data suggest that TLR4/9 may be involved in the development and progression of gastric cancer, and TLR4/9 may be potential target for gastric cancer treatment．
Objective:To investigate the correlation of Toll-like receptors（TLR）4 and TLR9 expression with the pathological stages and metastases of gastric cancer. Methods: Forty-two gastric cancer and paracancerous tissues were included in the present study (patients were diagnosed in No.174 Hospital of PLA from Jan.1 2009 to Oct. 2009). The mRNA and protein expressions of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 in the gastric cancer and paracancerous tissues were detected by RT-PCR and Western blotting analysis, respectively. Results: The expressions of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 mRNA were significantly higher in the gastric cancer tissues than those in the paracancerous tissues (1.29±0.03 vs 0.53±0.02, 0.99±0.04 vs 0.22±0.05；P<0.05). T3+T4 gastric cancer tissues demonstrated significantly higher levels of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 mRNA than the T1+T2 tissues (P<0.05). Significantly higher levels of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 mRNA were observed in patients with distant metastases than in those without (P<0.05). However, no correlation was observed between 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 mRNA expression with the age, gender or lymphatic metastasis status of patients. The expressions of 〖STBX〗TLR4〖STBZ〗 and 〖STBX〗TLR9〖STBZ〗 protein were significantly higher in the gastric cancer tissues than those in the paracancerous tissues (1.36±0.05 vs 0.48±0.04, 112±0.05 vs 0.34±0.03;P<0.05), T3+T4 gastric cancer tissues demonstrated significantly higher levels of TLR4 and TLR9 proteins than the T1+T2 tissues (P<0.05). Conclusions: TLR4 and TLR9 are up-regulated in gastric carcinoma tissues, and TLR4/9 expression is correlated with the TNM staging and distant metastasis. These data suggest that TLR4/9 may be involved in the development and progression of gastric cancer, and TLR4/9 may be potential target for gastric cancer treatment．
ZHANG Peng
,
HUANG Peng
,
GUO Xian-wen
,
LEI Rong-e
,
YUE Cheng-si
,
LING Min
,
LIN Wen-zhen
,
CAI Dan-zhao
,
HE Su-jia
,
TANG Wen-jue
,
ZHOU Su-fang
2011, 18(2):206-210.
DOI: 10.3872/j.issn.1007-385X.2011.2.017
Abstract:
Objective:To explore the expression of S100 calcium binding protein A13 (S100A13) in hepatocellular carcinoma (HCC) tissues and its clinical significance. Methods: Thirty-four HCC tissues, 18 corresponding paracancerous tissues, and 9 normal liver tissues were obtained from First Affiliated Hospital of Guangxi Medical University, and the high density chips of 197 HCC tissues and corresponding paracancerous tissues were obtained from commercial company. In situ RT-PCR was used to detect the expression of S100A13 mRNA in HCC tissues, paracancerous tissues, and normal liver tissues. Immunohistochemistry was used to detect the expression of S100A13 protein in HCC tissues, paracancerous tissues, and normal liver tissues. We analyzed the optical density of immunohistochemistry results by Image-Pro Plus. Results: The expression rates of S100A13 mRNA in HCC tissues, their corresponding paracancerous tissues and normal liver tissues were 70.21%, 51.06%, and 33.33%, respectively. S100A13 mRNA was mainly expressed in the nucleus, especially in nuclear membranes. The expression rates of S100A13 protein in HCC tissues, their corresponding paracancerous tissues and normal liver tissues were 55.84%, 38.96%, and 26.32%, respectively. The highest average optical density was found in HCC tissue (0.038±0.051), followed by paracancerous tissues (0.022±0.034), and the lowest was found in normal liver tissues (0.01±0.009) (P＜0.05). S100A13 protein was mainly expressed in the cytoplasm of HCC cells, occasionally in the nucleus; the secondary bile canaliculi and some inflammatory cells also showed high S100A13 protein expression. There was no correlation between S100A13 expression and the sex, age, and tumor grade of HCC patients. Conclusion: S100A13 protein is highly expressed in HCC, and it might serve as a potential target in HCC therapy.
Objective:To explore the expression of S100 calcium binding protein A13 (S100A13) in hepatocellular carcinoma (HCC) tissues and its clinical significance. Methods: Thirty-four HCC tissues, 18 corresponding paracancerous tissues, and 9 normal liver tissues were obtained from First Affiliated Hospital of Guangxi Medical University, and the high density chips of 197 HCC tissues and corresponding paracancerous tissues were obtained from commercial company. In situ RT-PCR was used to detect the expression of S100A13 mRNA in HCC tissues, paracancerous tissues, and normal liver tissues. Immunohistochemistry was used to detect the expression of S100A13 protein in HCC tissues, paracancerous tissues, and normal liver tissues. We analyzed the optical density of immunohistochemistry results by Image-Pro Plus. Results: The expression rates of S100A13 mRNA in HCC tissues, their corresponding paracancerous tissues and normal liver tissues were 70.21%, 51.06%, and 33.33%, respectively. S100A13 mRNA was mainly expressed in the nucleus, especially in nuclear membranes. The expression rates of S100A13 protein in HCC tissues, their corresponding paracancerous tissues and normal liver tissues were 55.84%, 38.96%, and 26.32%, respectively. The highest average optical density was found in HCC tissue (0.038±0.051), followed by paracancerous tissues (0.022±0.034), and the lowest was found in normal liver tissues (0.01±0.009) (P＜0.05). S100A13 protein was mainly expressed in the cytoplasm of HCC cells, occasionally in the nucleus; the secondary bile canaliculi and some inflammatory cells also showed high S100A13 protein expression. There was no correlation between S100A13 expression and the sex, age, and tumor grade of HCC patients. Conclusion: S100A13 protein is highly expressed in HCC, and it might serve as a potential target in HCC therapy.
17 Expressions of 14-3-3σ and cyclin B1 proteins in breast carcinoma and their clinical significances
2011, 18(2):211-215.
DOI: 10.3872/j.issn.1007-385X.2011.2.018
Abstract:
Objective:To study the expressions of 14-3-3σ and cyclin B1 proteins in breast carcinoma tissues and to analyze their relationships with clinical characteristics of breast carcinoma. Methods: Ninety-five breast carcinoma tissues and thirty corresponding adjacent tissues (patients were diagnosed in First Affiliated Hospital of Liaoning Medical University from Dec. 2006 to Dec. 2009; 31〖KG-*2〗〖DK〗-85 years old, median age 58 years) were included in the present study. The expressions of 14-3-3σ and cyclin B1 proteins in the breast carcinoma tissues and adjacent tissues were detected by immunohistochemistry PV and Western blotting methods, and their correlation with clinical characteristics of breast carcinoma was analyzed. Results: Immunohistochemistry and Western blotting results showed that the expression rate of 14-3-3σ in breast carcinoma was 22.1% (21/95), which was significantly lower than that in the adjacent tissues (93.3%, 28/30; P<005); the expression of 14-3-3σ was correlated with the histological grade, TNM stage, and lymph node metastasis of breast carcinoma (P<0.05). The expression rates of cyclin B1 in breast carcinoma was 69.5% (66/95), being significantly higher than that in the adjacent tissues (40%, 12/30; P<0.05). The expression of cyclin B1 was only correlated with lymph node metastasis of breast carcinoma (P<005); expression of 14-3-3σ was negatively correlated with cyclin B1 in the breast carcinoma tissues (r=-0.333, P=0.001). Conclusion: 14-3-3σ and cyclin B1 proteins are related to the progression and metastasis of breast carcinoma, and their combined detection may help the diagnosis and treatment of breast carcinoma.
Objective:To study the expressions of 14-3-3σ and cyclin B1 proteins in breast carcinoma tissues and to analyze their relationships with clinical characteristics of breast carcinoma. Methods: Ninety-five breast carcinoma tissues and thirty corresponding adjacent tissues (patients were diagnosed in First Affiliated Hospital of Liaoning Medical University from Dec. 2006 to Dec. 2009; 31〖KG-*2〗〖DK〗-85 years old, median age 58 years) were included in the present study. The expressions of 14-3-3σ and cyclin B1 proteins in the breast carcinoma tissues and adjacent tissues were detected by immunohistochemistry PV and Western blotting methods, and their correlation with clinical characteristics of breast carcinoma was analyzed. Results: Immunohistochemistry and Western blotting results showed that the expression rate of 14-3-3σ in breast carcinoma was 22.1% (21/95), which was significantly lower than that in the adjacent tissues (93.3%, 28/30; P<005); the expression of 14-3-3σ was correlated with the histological grade, TNM stage, and lymph node metastasis of breast carcinoma (P<0.05). The expression rates of cyclin B1 in breast carcinoma was 69.5% (66/95), being significantly higher than that in the adjacent tissues (40%, 12/30; P<0.05). The expression of cyclin B1 was only correlated with lymph node metastasis of breast carcinoma (P<005); expression of 14-3-3σ was negatively correlated with cyclin B1 in the breast carcinoma tissues (r=-0.333, P=0.001). Conclusion: 14-3-3σ and cyclin B1 proteins are related to the progression and metastasis of breast carcinoma, and their combined detection may help the diagnosis and treatment of breast carcinoma.
2011, 18(2):216-219.
DOI: 10.3872/j.issn.1007-385X.2011.2.019
Abstract:
Objective:To investigate the expression and clinical significance of IL-17 in primary human hepatocellular carcinoma (HCC). Methods: In the present study 23 primary HCC patients and 9 benign liver tumor patients were collected from the First Affiliated Hospital of Nanjing Medical University, with the tumor tissues and paracancerous tissues obtained by surgery. The serum IL-17 level was measured by ELISA, the expression of IL-17, ROR-γt mRNA in HCC tissues were detected by real-time PCR, and the protein expression of IL-17 in HCC tissues was examined by immunohistochemistry method. Results: The serum IL-17 level was greatly higher in HCC patients compared with that in benign liver tumor patients. Real-time PCR results showed that the expression levels of IL-17, ROR-γt mRNA were significantly higher in cancer tissues than in the peritumoral and normal liver tissues (P<005). Immunohistochemistry results showed that the expression of IL-17 protein was greatly higher in the cancer and peritumoral tissues than that in the normal liver tissues, and that in the cancer tissues was higher than that in the peritumoral tissues. Conclusion: IL-17 is highly expressed in HCC tissues, which may provide a new target in the diagnosis and therapy of HCC.
Objective:To investigate the expression and clinical significance of IL-17 in primary human hepatocellular carcinoma (HCC). Methods: In the present study 23 primary HCC patients and 9 benign liver tumor patients were collected from the First Affiliated Hospital of Nanjing Medical University, with the tumor tissues and paracancerous tissues obtained by surgery. The serum IL-17 level was measured by ELISA, the expression of IL-17, ROR-γt mRNA in HCC tissues were detected by real-time PCR, and the protein expression of IL-17 in HCC tissues was examined by immunohistochemistry method. Results: The serum IL-17 level was greatly higher in HCC patients compared with that in benign liver tumor patients. Real-time PCR results showed that the expression levels of IL-17, ROR-γt mRNA were significantly higher in cancer tissues than in the peritumoral and normal liver tissues (P<005). Immunohistochemistry results showed that the expression of IL-17 protein was greatly higher in the cancer and peritumoral tissues than that in the normal liver tissues, and that in the cancer tissues was higher than that in the peritumoral tissues. Conclusion: IL-17 is highly expressed in HCC tissues, which may provide a new target in the diagnosis and therapy of HCC.
19 Anti-cancer effect and its mechanism of human α-lactalbumin made lethal to tumor cells: An advance
2011, 18(2):220-224.
DOI: 10.3872/j.issn.1007-385X.2011.2.020
Abstract:
肿瘤细胞致死性人α-乳清蛋白（human α-lactalbumin made lethal to tumor cells，HAMLET）是人α-乳清蛋白与油酸结合后改变自身三级结构而形成的脂蛋白复合物，能够选择性杀伤肿瘤细胞，而对正常细胞几乎无影响。HAMLET杀伤肿瘤细胞的机制目前还未完全明确。早期研究显示，HAMLET通过细胞凋亡机制杀伤肿瘤细胞；进一步研究发现，HAMLET并不主要依赖凋亡，而是通过细胞自噬、内质网应激、蛋白酶体、溶酶体、组蛋白和DAN等多种途径杀伤肿瘤细胞。HAMLET对皮肤乳头状瘤、膀胱癌局部用药的抗肿瘤临床试验已取得良好的效果， 它很可能成为具有良好应用前景的绿色抗肿瘤新药。
肿瘤细胞致死性人α-乳清蛋白（human α-lactalbumin made lethal to tumor cells，HAMLET）是人α-乳清蛋白与油酸结合后改变自身三级结构而形成的脂蛋白复合物，能够选择性杀伤肿瘤细胞，而对正常细胞几乎无影响。HAMLET杀伤肿瘤细胞的机制目前还未完全明确。早期研究显示，HAMLET通过细胞凋亡机制杀伤肿瘤细胞；进一步研究发现，HAMLET并不主要依赖凋亡，而是通过细胞自噬、内质网应激、蛋白酶体、溶酶体、组蛋白和DAN等多种途径杀伤肿瘤细胞。HAMLET对皮肤乳头状瘤、膀胱癌局部用药的抗肿瘤临床试验已取得良好的效果， 它很可能成为具有良好应用前景的绿色抗肿瘤新药。
2011, 18(2):225-229.
DOI: 10.3872/j.issn.1007-385X.2011.2.021
Abstract:
芳香烃受体（aryl hydrocarbon receptor，AhR）又名二噁英受体，是一种配体激活性转录因子，当与多环芳烃、卤代芳烃等配体结合后，可调控一系列基因的表达。AhR除了参与外源化合物的代谢外，还参与调控许多重要的生物学过程，包括个体发育、细胞分化、癌变等。AhR 高表达于乳腺癌、肺癌、胰腺癌和胃癌等多种肿瘤中，可调控肿瘤细胞的增殖、周期、凋亡，以及细胞迁移和侵袭, 并且在不同的细胞、不同的环境所起的作用不同。AhR的某些配体（3,3′-吲哚甲烷，tranilast等）显示有抑制肿瘤细胞生长、预防肿瘤发生的作用，因此AhR有望成为肿瘤治疗的新靶点。
芳香烃受体（aryl hydrocarbon receptor，AhR）又名二噁英受体，是一种配体激活性转录因子，当与多环芳烃、卤代芳烃等配体结合后，可调控一系列基因的表达。AhR除了参与外源化合物的代谢外，还参与调控许多重要的生物学过程，包括个体发育、细胞分化、癌变等。AhR 高表达于乳腺癌、肺癌、胰腺癌和胃癌等多种肿瘤中，可调控肿瘤细胞的增殖、周期、凋亡，以及细胞迁移和侵袭, 并且在不同的细胞、不同的环境所起的作用不同。AhR的某些配体（3,3′-吲哚甲烷，tranilast等）显示有抑制肿瘤细胞生长、预防肿瘤发生的作用，因此AhR有望成为肿瘤治疗的新靶点。
2011, 18(2):230-234.
DOI: 10.3872/j.issn.1007-385X.2011.2.022
Abstract:
信号转导及转录活化因子3（signal transducer and activator of transcription 3，STAT3）是STATs家族中的重要成员，其在多种肿瘤细胞中持续激活并过度表达，与肿瘤细胞的增殖、侵袭、转移等密切相关，已成为肿瘤治疗的热点靶分子之一。目前已发现的针对STAT3的抑制剂主要有核酸类抑制剂、蛋白类抑制剂以及小分子化合物抑制剂等，其中靶向STAT3核酸类抑制剂根据其作用方式可分为小分子干扰RNA（small interfering RNA，siRNA）、反义寡核苷酸（antisense oligonucleotide，AS-ON）、诱饵寡核苷酸（decoy oligonucleotide，Decoy ON）、G-四联体寡聚脱氧核苷酸（G-quartet oligodeoxynucleotides，GQ-ODN）和显性负性质粒（dominant-negative plasmids）等5种。这些核酸类抑制剂能够在DNA或RNA水平上，通过多种方式影响STAT3活性，抑制其下游信号分子的表达，发挥抗肿瘤的活性。此类抑制剂具有作用位点明确、特异性强以及可操作性强等优点，已成为分子靶向治疗肿瘤研究中不可或缺的工具，具有潜在的临床应用价值。
信号转导及转录活化因子3（signal transducer and activator of transcription 3，STAT3）是STATs家族中的重要成员，其在多种肿瘤细胞中持续激活并过度表达，与肿瘤细胞的增殖、侵袭、转移等密切相关，已成为肿瘤治疗的热点靶分子之一。目前已发现的针对STAT3的抑制剂主要有核酸类抑制剂、蛋白类抑制剂以及小分子化合物抑制剂等，其中靶向STAT3核酸类抑制剂根据其作用方式可分为小分子干扰RNA（small interfering RNA，siRNA）、反义寡核苷酸（antisense oligonucleotide，AS-ON）、诱饵寡核苷酸（decoy oligonucleotide，Decoy ON）、G-四联体寡聚脱氧核苷酸（G-quartet oligodeoxynucleotides，GQ-ODN）和显性负性质粒（dominant-negative plasmids）等5种。这些核酸类抑制剂能够在DNA或RNA水平上，通过多种方式影响STAT3活性，抑制其下游信号分子的表达，发挥抗肿瘤的活性。此类抑制剂具有作用位点明确、特异性强以及可操作性强等优点，已成为分子靶向治疗肿瘤研究中不可或缺的工具，具有潜在的临床应用价值。
2011, 18(2):235-238.
DOI: 10.3872/j.issn.1007-385X.2011.2.023
Abstract:
微小RNA（microRNA，miRNA）通过调控基因的表达，参与细胞生命过程中一系列重要的进程，包括胚胎发育、细胞增殖和分化、细胞死亡与凋亡、体内生化代谢等。成熟miRNA通过RNA诱导的沉默复合体（RNA-induced silencing complex，RISC）结合到靶mRNA上，依赖于序列的互补性机制、剪切或阻遏靶mRNA、沉默基因的表达。miRNA与肿瘤等疾病的发生、发展密切相关。miRNA的表达谱在肿瘤细胞与正常细胞之间具有明显差异，起到类似于癌基因或抑癌基因的作用。miRNA通过沉默肿瘤侵袭转移相关基因的表达，参与肿瘤侵袭转移过程。肿瘤细胞在miRNA表达谱上的特异性为肿瘤的诊断提供了一项生物标志物，同时也为调控miRNA表达以治疗肿瘤提供了新的靶点。以miRNA为基础的抗肿瘤治疗还可与传统的化疗结合起来，提高肿瘤的治疗效果，为肿瘤的生物治疗开拓新视野。
微小RNA（microRNA，miRNA）通过调控基因的表达，参与细胞生命过程中一系列重要的进程，包括胚胎发育、细胞增殖和分化、细胞死亡与凋亡、体内生化代谢等。成熟miRNA通过RNA诱导的沉默复合体（RNA-induced silencing complex，RISC）结合到靶mRNA上，依赖于序列的互补性机制、剪切或阻遏靶mRNA、沉默基因的表达。miRNA与肿瘤等疾病的发生、发展密切相关。miRNA的表达谱在肿瘤细胞与正常细胞之间具有明显差异，起到类似于癌基因或抑癌基因的作用。miRNA通过沉默肿瘤侵袭转移相关基因的表达，参与肿瘤侵袭转移过程。肿瘤细胞在miRNA表达谱上的特异性为肿瘤的诊断提供了一项生物标志物，同时也为调控miRNA表达以治疗肿瘤提供了新的靶点。以miRNA为基础的抗肿瘤治疗还可与传统的化疗结合起来，提高肿瘤的治疗效果，为肿瘤的生物治疗开拓新视野。
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.