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Volume 19,Issue 5,2012 Table of Contents
2012, 19(5):467-471.
DOI: 10.3872/j.issn.1007-385X.2012.5.001
Abstract:
In-depth study of the molecular mechanisms of lung adenocarcinoma resistance to docetaxel (DTX) not only offers us the new promising targets of individualized treatment for lung adenocarcinoma patients, but also provides new insights into clinical intervention strategies. In this study, the docetaxel-resistant human lung adenocarcinoma cell line SPC-A1/DTX was derived from parental SPC-A1 cell line by continuous exposure to increasing concentration of DTX in vitro. Differences in biological characteristics, such as cell morphology, chemosensitivity, cell proliferation, apoptosis, cell cycle, and drug transportation were compared between SPC-A1 and SPC-A1/DTX cell lines. Based on gene expression- and miRNA-microarray analysis, differentially expressed genes, such as ING4 and miRNAs (e.g. miR-200b and miR-100) were screened out from SPC-A1/DTX cells. With further gain-of-function and (or) loss-of-function studies of these molecules in both in vitro and in vivo models, we provided potential mechanistic explanations for DTX resistance of human lung adenocarcinoma. It was found that down-regulation of ING4 gene and abnormalities of crosstalk between miR-200b and E2F3gene and crosstalk between miR-100 and Plk-1gene might be essential for DTX resistance in human lung adenocarcinoma.
In-depth study of the molecular mechanisms of lung adenocarcinoma resistance to docetaxel (DTX) not only offers us the new promising targets of individualized treatment for lung adenocarcinoma patients, but also provides new insights into clinical intervention strategies. In this study, the docetaxel-resistant human lung adenocarcinoma cell line SPC-A1/DTX was derived from parental SPC-A1 cell line by continuous exposure to increasing concentration of DTX in vitro. Differences in biological characteristics, such as cell morphology, chemosensitivity, cell proliferation, apoptosis, cell cycle, and drug transportation were compared between SPC-A1 and SPC-A1/DTX cell lines. Based on gene expression- and miRNA-microarray analysis, differentially expressed genes, such as ING4 and miRNAs (e.g. miR-200b and miR-100) were screened out from SPC-A1/DTX cells. With further gain-of-function and (or) loss-of-function studies of these molecules in both in vitro and in vivo models, we provided potential mechanistic explanations for DTX resistance of human lung adenocarcinoma. It was found that down-regulation of ING4 gene and abnormalities of crosstalk between miR-200b and E2F3gene and crosstalk between miR-100 and Plk-1gene might be essential for DTX resistance in human lung adenocarcinoma.
2012, 19(5):472-477.
DOI: 10.3872/j.issn.1007-385X.2012.5.002
Abstract:
Objective:To explore the cytotoxicity and the underlying mechanisms of peripheral blood mononuclear cells (PBMCs) activated by IL-2 and IL-15 on human Burkitt lymphoma Raji cells after being treated by norcantharidin (NCTD).Methods: Trypan blue assay was used to detect the inhibitory effect of NCTD on Raji cells and PBMC. The cytotoxic effects of PBMC activated by IL-2 and IL-15 against K562 cells and Raji cells were analyzed by LDH releasing assay. The expression of NKG2D on the surface of PBMC activated by IL-2 and IL-15 was assayed by flow cytometry (FCM). The expressions of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) on Raji cells were assayed by FCM before and after NCTD treatment. Results: NCTD inhibited the proliferation of Raji cells significantly in a dose- and time-dependent manner (P<0.05), on the contrary, it showed no significant effect on PBMC proliferation (P>0.05). The cytotoxic rates of IL-2 and IL-15 activated PBMC on K562 cells were more than that of inactivated PBMC when the ratio of effect/target (E∶T) were 10∶1 and 20∶1 (\[52.42±3.89\]% vs \[15.82±5.12\]%, \[79.55±922\]% vs \[2767±366\]%, P<0.05), respectively. The cytotoxic rates of inactivated PBMC against Raji cells after being treated by NCTD were increased compared with untreated Raji cells (\[23.63±6.20\]% vs \[5.04±1.25\]%, \[41.80±409\]% vs \[8.59±2.19\]%, P<0.05) with E∶T of 10∶1 and 20∶1 respectively. Meanwhile, the cytotoxic rates of PBMC after being activated by IL-2 and IL-15 on NCTD-treated Raji cells were also increased compared with that of untreated PBMC (\[38.97±2.76\]% vs \[13.19±3.67\]%, \[63.09±7.30\]% vs \[19.89±4.15\]%, P<0.05). The expression of NKG2D on PBMC activated by IL-2 and IL-15 was increased compared with untreated PBMC (\[4491±585\]% vs \[25.28±769\]%, P<0.05). The expressions of NKG2D ligands on Raji cells, especially ULBP2 were up-regulated (\[12.69±3.99\]% vs \[1.03±0.42\]%, P<0.05) after being treated by NCTD, while no significant changes were found in other ligands (P>0.05). Conclusion: Combining NCTD with IL-2 and IL-15 can increase the cytotoxicity of PBMC on Raji cells, which may be related with NCTD-increased expression of ULBP2 on Raji cell surface and cytokine-increased expression of NKG2D on PBMC surface.
Objective:To explore the cytotoxicity and the underlying mechanisms of peripheral blood mononuclear cells (PBMCs) activated by IL-2 and IL-15 on human Burkitt lymphoma Raji cells after being treated by norcantharidin (NCTD).Methods: Trypan blue assay was used to detect the inhibitory effect of NCTD on Raji cells and PBMC. The cytotoxic effects of PBMC activated by IL-2 and IL-15 against K562 cells and Raji cells were analyzed by LDH releasing assay. The expression of NKG2D on the surface of PBMC activated by IL-2 and IL-15 was assayed by flow cytometry (FCM). The expressions of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) on Raji cells were assayed by FCM before and after NCTD treatment. Results: NCTD inhibited the proliferation of Raji cells significantly in a dose- and time-dependent manner (P<0.05), on the contrary, it showed no significant effect on PBMC proliferation (P>0.05). The cytotoxic rates of IL-2 and IL-15 activated PBMC on K562 cells were more than that of inactivated PBMC when the ratio of effect/target (E∶T) were 10∶1 and 20∶1 (\[52.42±3.89\]% vs \[15.82±5.12\]%, \[79.55±922\]% vs \[2767±366\]%, P<0.05), respectively. The cytotoxic rates of inactivated PBMC against Raji cells after being treated by NCTD were increased compared with untreated Raji cells (\[23.63±6.20\]% vs \[5.04±1.25\]%, \[41.80±409\]% vs \[8.59±2.19\]%, P<0.05) with E∶T of 10∶1 and 20∶1 respectively. Meanwhile, the cytotoxic rates of PBMC after being activated by IL-2 and IL-15 on NCTD-treated Raji cells were also increased compared with that of untreated PBMC (\[38.97±2.76\]% vs \[13.19±3.67\]%, \[63.09±7.30\]% vs \[19.89±4.15\]%, P<0.05). The expression of NKG2D on PBMC activated by IL-2 and IL-15 was increased compared with untreated PBMC (\[4491±585\]% vs \[25.28±769\]%, P<0.05). The expressions of NKG2D ligands on Raji cells, especially ULBP2 were up-regulated (\[12.69±3.99\]% vs \[1.03±0.42\]%, P<0.05) after being treated by NCTD, while no significant changes were found in other ligands (P>0.05). Conclusion: Combining NCTD with IL-2 and IL-15 can increase the cytotoxicity of PBMC on Raji cells, which may be related with NCTD-increased expression of ULBP2 on Raji cell surface and cytokine-increased expression of NKG2D on PBMC surface.
2012, 19(5):478-485.
DOI: 10.3872/j.issn.1007-385X.2012.5.003
Abstract:
Objective:To explore the possibility of enriching breast cancer stem cell-like cells by cell passage in a tumor-bearing mouse model treated with 5-fluorouracil （5-FU） in vivo and to lay a foundation for cancer stem cell-targeted therapy. Methods: To establish a tumor-bearing model by inoculating breast cancer cell line 4T1 into mouse subcutaneously. 5-FU was injected into these mice intraperitonealy according to different groups. The mice were executed 4 weeks after injection. Cell suspension made from mouse tumor was injected into the mice again to establish the next generation of tumor-bearing mouse model. The mice were treated with 5-FU in the third and fourth generation as above. Four generations of the mouse model were established in all. The mice in control group were injected with normal saline instead of 5-FU, and the following four generation tumor-bearing models were treated as 5-FU group. Flow cytometry was used to measure the proportion of CD44+CD24-/low cells in different tumor tissues; Hoechast 33342 staining was used to measure the proportion of SP (side population) cells; and expressions of ALDH1 and CD55 proteins were detected by immunohistochemistry. The formation of mammospheres was observed under an inverted microscope. Tumorigenic ability of different tumor cells was detected by mouse tumorigenesis experiment. Results: In the control group, the proportion of CD44+CD24-/lowcells in mouse tumor was (11.5±0.9)%, and the proportion of SP cells was (9.7±13)%. The expression level of ALDH1 was negative, the proportion of cells in which CD55 was highly expressed, was (0.6±0.3)%, and the proportion of mammospheres was (0.5±0.2)%. While in 5-FU treated group, the proportions of CD44+CD24-/lowcells were respectively (49.8±1.2)%, (56.8±1.7)%, (66.4±1.5)% and (69.0±1.6)%; the proportions of SP cells were respectively (25.0±1.2)%, (42.6±2.8)%, (58.4±2.1)% and (61.3±2.6)%; the proportions of cells in which ALDH1 was positively expressed, were respectively (3.8±0.7)%, (14.1±24)%, (25.2±31)% and (275±2.7)%, and the proportions of cells in which CD55 were highly expressed, were respectively (7.8±16)%, (10.1±2.0)%, (15.6±1.4)% and (17.3±1.9)%, and the proportion of mammospheres was (5.9±04)%. The significant differences were found between the two groups（P<0.05 or P<0.01）. The third generation tumor cells enriching cancer stem cells were more tumorigenic than the control tumor cells. Conclusion: 5-FU enrichs the cancer stem cell-like cells in mouse breast cancer cell line 4T1 by the cell passage in the tumor-bearing mouse model in vivo.
Objective:To explore the possibility of enriching breast cancer stem cell-like cells by cell passage in a tumor-bearing mouse model treated with 5-fluorouracil （5-FU） in vivo and to lay a foundation for cancer stem cell-targeted therapy. Methods: To establish a tumor-bearing model by inoculating breast cancer cell line 4T1 into mouse subcutaneously. 5-FU was injected into these mice intraperitonealy according to different groups. The mice were executed 4 weeks after injection. Cell suspension made from mouse tumor was injected into the mice again to establish the next generation of tumor-bearing mouse model. The mice were treated with 5-FU in the third and fourth generation as above. Four generations of the mouse model were established in all. The mice in control group were injected with normal saline instead of 5-FU, and the following four generation tumor-bearing models were treated as 5-FU group. Flow cytometry was used to measure the proportion of CD44+CD24-/low cells in different tumor tissues; Hoechast 33342 staining was used to measure the proportion of SP (side population) cells; and expressions of ALDH1 and CD55 proteins were detected by immunohistochemistry. The formation of mammospheres was observed under an inverted microscope. Tumorigenic ability of different tumor cells was detected by mouse tumorigenesis experiment. Results: In the control group, the proportion of CD44+CD24-/lowcells in mouse tumor was (11.5±0.9)%, and the proportion of SP cells was (9.7±13)%. The expression level of ALDH1 was negative, the proportion of cells in which CD55 was highly expressed, was (0.6±0.3)%, and the proportion of mammospheres was (0.5±0.2)%. While in 5-FU treated group, the proportions of CD44+CD24-/lowcells were respectively (49.8±1.2)%, (56.8±1.7)%, (66.4±1.5)% and (69.0±1.6)%; the proportions of SP cells were respectively (25.0±1.2)%, (42.6±2.8)%, (58.4±2.1)% and (61.3±2.6)%; the proportions of cells in which ALDH1 was positively expressed, were respectively (3.8±0.7)%, (14.1±24)%, (25.2±31)% and (275±2.7)%, and the proportions of cells in which CD55 were highly expressed, were respectively (7.8±16)%, (10.1±2.0)%, (15.6±1.4)% and (17.3±1.9)%, and the proportion of mammospheres was (5.9±04)%. The significant differences were found between the two groups（P<0.05 or P<0.01）. The third generation tumor cells enriching cancer stem cells were more tumorigenic than the control tumor cells. Conclusion: 5-FU enrichs the cancer stem cell-like cells in mouse breast cancer cell line 4T1 by the cell passage in the tumor-bearing mouse model in vivo.
2012, 19(5):486-490.
DOI: 10.3872/j.issn.1007-385X.2012.5.004
Abstract:
Objective:To detect the expression of Rac1 protein in colorectal cancer cells and to observe its correlation with cytoskeleton, cell cycle and apoptosis of colorectal cancer LoVo cells. Methods: The expression of Rac1 protein was detected by Western blotting in five colorectal cancer cell lines. The cytoskeleton changes were observed by confocal microscopy in LoVo cells transfected with Rac1-shRNA. The cell cycle and apoptosis were observed by flow cytometry in LoVo cells transfected with Rac1-shRNA. Results: Rac1 protein was highly expressed in five colorectal cancer cells. The cross-linked F-actin network was significantly reduced and disordered in LoVo cells transfected with Rac1-shRNA. The cell number in G0/G1 phase was significantly increased in Rac1-shRNA group compared with the control group (\[74.63±440\]% vs \[56.46±3.09\]%，P<0.05), whereas it was reduced in S phase (\[12.87±1.77\]% vs \[29.66±192\]%，P<0.05) and the apoptosis rate of LoVo cells was significantly increased in Rac1-shRNA group compared with the control group (\[25.31±2.05\]% vs \[9.38±1.16\]%, P<0.05). Conclusion: Silencing Rac1 expression mediated by RNA interference affects the formation of cytoskeleton and the cell cycle of LoVo cells, which provides clinical evidence for Rac1-targeted treatment of colorectal cancer.
Objective:To detect the expression of Rac1 protein in colorectal cancer cells and to observe its correlation with cytoskeleton, cell cycle and apoptosis of colorectal cancer LoVo cells. Methods: The expression of Rac1 protein was detected by Western blotting in five colorectal cancer cell lines. The cytoskeleton changes were observed by confocal microscopy in LoVo cells transfected with Rac1-shRNA. The cell cycle and apoptosis were observed by flow cytometry in LoVo cells transfected with Rac1-shRNA. Results: Rac1 protein was highly expressed in five colorectal cancer cells. The cross-linked F-actin network was significantly reduced and disordered in LoVo cells transfected with Rac1-shRNA. The cell number in G0/G1 phase was significantly increased in Rac1-shRNA group compared with the control group (\[74.63±440\]% vs \[56.46±3.09\]%，P<0.05), whereas it was reduced in S phase (\[12.87±1.77\]% vs \[29.66±192\]%，P<0.05) and the apoptosis rate of LoVo cells was significantly increased in Rac1-shRNA group compared with the control group (\[25.31±2.05\]% vs \[9.38±1.16\]%, P<0.05). Conclusion: Silencing Rac1 expression mediated by RNA interference affects the formation of cytoskeleton and the cell cycle of LoVo cells, which provides clinical evidence for Rac1-targeted treatment of colorectal cancer.
2012, 19(5):491-495.
DOI: 10.3872/j.issn.1007-385X.2012.5.005
Abstract:
Objective : To construct a recombinant adenovirus for calreticulin (CRT)/melanoma antigen gene-A3 (MAGE-A3) and evaluate its inhibitory effect on breast cancer MDA-MB-231 cells. Methods: Ad-CRT/MAGE-A3 vector was constructed and infected into MDA-MB-231 cells. The optimal multiplicity of infection (MOI) was obtained by fluorescence microscope method. There were five groups: epirubicin, Ad-GFP, Ad-CRT/MAGE-A3, epirubicin+Ad-GFP and epirubicin+Ad-CRT/MAGE-A3. The proliferation and invasion of MDA-MB-231 cells in different groups were detected by MTT and Transwell assays. Results: Recombinant adenovirus vector Ad-CRT/MAGE-A3 was constructed successfully, and its optimal MOI in MDA-MB-231 cells was 100. Ad-CRT/MAGE-A3 combined with epirubicin decreased the proliferation (\[83.27±1.04\]% vs \[57.42±1.27\]%, \[43.26±0.95\]%, \[61.23±1.47\]%, \[55.38±1.62\]%, P<0.05) and the invasion (\[8.41±4.20\] vs \[14.62±5.33\], \[107.66±3.35\], \[100.60±442\], \[104.20±260\], P<0.05) of MDA-MB-231 cells significantly. Conclusion: Recombinant adenovirus vector Ad-CRT/MAGE-A3 can enhance epirubicin to inhibit the proliferation and invasion of breast cancer MDA-MB-231 cells.
Objective : To construct a recombinant adenovirus for calreticulin (CRT)/melanoma antigen gene-A3 (MAGE-A3) and evaluate its inhibitory effect on breast cancer MDA-MB-231 cells. Methods: Ad-CRT/MAGE-A3 vector was constructed and infected into MDA-MB-231 cells. The optimal multiplicity of infection (MOI) was obtained by fluorescence microscope method. There were five groups: epirubicin, Ad-GFP, Ad-CRT/MAGE-A3, epirubicin+Ad-GFP and epirubicin+Ad-CRT/MAGE-A3. The proliferation and invasion of MDA-MB-231 cells in different groups were detected by MTT and Transwell assays. Results: Recombinant adenovirus vector Ad-CRT/MAGE-A3 was constructed successfully, and its optimal MOI in MDA-MB-231 cells was 100. Ad-CRT/MAGE-A3 combined with epirubicin decreased the proliferation (\[83.27±1.04\]% vs \[57.42±1.27\]%, \[43.26±0.95\]%, \[61.23±1.47\]%, \[55.38±1.62\]%, P<0.05) and the invasion (\[8.41±4.20\] vs \[14.62±5.33\], \[107.66±3.35\], \[100.60±442\], \[104.20±260\], P<0.05) of MDA-MB-231 cells significantly. Conclusion: Recombinant adenovirus vector Ad-CRT/MAGE-A3 can enhance epirubicin to inhibit the proliferation and invasion of breast cancer MDA-MB-231 cells.
2012, 19(5):496-501.
DOI: 10.3872/j.issn.1007-385X.2012.5.006
Abstract:
Objective : To construct two recombinant adenovirus vectors carrying reporter gene enhanced green fluorescent protein (EGFP) or suicide gene herpes simple virus thymidine kinase (HSV-TK) at the downstream of enhancer and promoter of human mammaglobin (hMAM-EP). To explore breast-cancer-cell-specific regulation effect of hMAM-EP and new methods of targeted therapy for breast cancer. Methods: Two recombinant plasmid vectors, hMAM-EP-EGFP and hMAM-EP-TK, were constructed, which respectively carried reporter gene EGFP and suicide gene HSV-TK at the downstream of hMAM-EP. Recombinant adenovirus vectors Ad-EP-EGFP and Ad-EP-TK were obtained after the target genes from the recombinant plasmids were transferred into adenovirus skeleton cosmid pAxcwit2; recombinant adenovirus vectors Ad-EP-EGFP and Ad-EP-TK were then transfected into breast cancer T-47D cells, ZR-75-30 cells and nasopharyngeal cancer 5-8F cells. The expression of EGFP was observed under a fluorescence microscope. Recombinant adenovirus Ad-EP-TK-infected T-47D cells were cultured with 1, 10, 20 and 50 μg/ml prodrug GCV to observe specific cell-killing effect on breast cancer cells. Results: The recombinant plasmid vectors Ad-EP-EGFP and Ad-EP-TK controlled by hMAM-EP were successfully constructed. EGFP could be observed in human breast cancer T-47D cells infected with Ad-EP-EGFP recombinant adenovirus, and could not be detected in ZR-75-30 and 5-8F cells. Compared with un-infected and Ad-EP-EGFP-infected groups, the survival rate of T-47D cells in Ad-EP-EGFP-infection combined with GCV (50 μg/ml) group was significantly decreased (\[35.69±0.07\]% vs \[91.74±0.02\]%, \[87.69±011\]%, P＜0.05). With an increase in mass concentration of GCV, the survival rate decreased. Cell survival rates were (94.34±0.04)%, (86.26±0.02)%, (66.51±0.09)% and (35.69±0.07)% when T-47D cells were infected with hMAM-EP-TK in a MOI of 100 and cultured with 1, 10, 20, and 50 μg/ml GCV. Conclusion: HSV-TK suicide gene controlled by hMAM-EP is specifically expressed in breast cancer T-47D cells, and T-47D cells can be killed by Ad-EP-TK combined with GCV.
Objective : To construct two recombinant adenovirus vectors carrying reporter gene enhanced green fluorescent protein (EGFP) or suicide gene herpes simple virus thymidine kinase (HSV-TK) at the downstream of enhancer and promoter of human mammaglobin (hMAM-EP). To explore breast-cancer-cell-specific regulation effect of hMAM-EP and new methods of targeted therapy for breast cancer. Methods: Two recombinant plasmid vectors, hMAM-EP-EGFP and hMAM-EP-TK, were constructed, which respectively carried reporter gene EGFP and suicide gene HSV-TK at the downstream of hMAM-EP. Recombinant adenovirus vectors Ad-EP-EGFP and Ad-EP-TK were obtained after the target genes from the recombinant plasmids were transferred into adenovirus skeleton cosmid pAxcwit2; recombinant adenovirus vectors Ad-EP-EGFP and Ad-EP-TK were then transfected into breast cancer T-47D cells, ZR-75-30 cells and nasopharyngeal cancer 5-8F cells. The expression of EGFP was observed under a fluorescence microscope. Recombinant adenovirus Ad-EP-TK-infected T-47D cells were cultured with 1, 10, 20 and 50 μg/ml prodrug GCV to observe specific cell-killing effect on breast cancer cells. Results: The recombinant plasmid vectors Ad-EP-EGFP and Ad-EP-TK controlled by hMAM-EP were successfully constructed. EGFP could be observed in human breast cancer T-47D cells infected with Ad-EP-EGFP recombinant adenovirus, and could not be detected in ZR-75-30 and 5-8F cells. Compared with un-infected and Ad-EP-EGFP-infected groups, the survival rate of T-47D cells in Ad-EP-EGFP-infection combined with GCV (50 μg/ml) group was significantly decreased (\[35.69±0.07\]% vs \[91.74±0.02\]%, \[87.69±011\]%, P＜0.05). With an increase in mass concentration of GCV, the survival rate decreased. Cell survival rates were (94.34±0.04)%, (86.26±0.02)%, (66.51±0.09)% and (35.69±0.07)% when T-47D cells were infected with hMAM-EP-TK in a MOI of 100 and cultured with 1, 10, 20, and 50 μg/ml GCV. Conclusion: HSV-TK suicide gene controlled by hMAM-EP is specifically expressed in breast cancer T-47D cells, and T-47D cells can be killed by Ad-EP-TK combined with GCV.
MAO Lian-gang
,
LI Ke-qiang
,
ZHUO Wen-ying
,
DAI Xiao-yu
,
YU Yong-ming
,
FENG Wei-yun
,
LE Dong-hai
2012, 19(5):502-507.
DOI: 10.3872/j.issn.1007-385X.2012.5.007
Abstract:
Objective : To investigate the effect of receptor tyrosine kinase recepteur d’origine nantais (RON) gene silencing on the invasion and anticancer drug resistance of human colon carcinoma HT-29 cells. Methods: RNAi lentiviral vector targeting RON gene (Lv-RON-siRNA) was constructed. The efficiency of Lv-RON-siRNA on RON gene silence and RON protein level in HT-29 cells were detected by real-time PCR and Western blotting, respectively. The effects of Lv-RON-siRNA on invasion and drug resistance of HT-29 cells were observed by Transwell assay and ATP-TCA (ATP-tumor chemosensitivity assay). Results: The silencing effect of Lv-RON-siRNA on RON gene expression in HT-29 cells reached 70%. Compared with the control group, the invasion of HT-29 cells in Lv-RON-siRNA infection group was decreased（097±0.07 vs 1.29±0.08, P<0.05). The values of IC90 and IC50 of HT-29 cells infected with Lv-RON-siRNA to 5-FU were (14.28±1.34) μg/ml and (8.93±1.2) μg/ml, respectively. The IC90 and IC50 of HT-29 cells infected with Lv-RON-siRNA to cisplatin (DDP) were (1.91±0.22) μg/ml and (0.64±0.07) μg/ml, respectively, and were significantly lower than those in the control group (P<0.01). Conclusion: Silencing RON gene expression can decrease the invasion ability of HT-29 cells and increase the sensitivity of HT-29 cells to 5-FU and DDP.
Objective : To investigate the effect of receptor tyrosine kinase recepteur d’origine nantais (RON) gene silencing on the invasion and anticancer drug resistance of human colon carcinoma HT-29 cells. Methods: RNAi lentiviral vector targeting RON gene (Lv-RON-siRNA) was constructed. The efficiency of Lv-RON-siRNA on RON gene silence and RON protein level in HT-29 cells were detected by real-time PCR and Western blotting, respectively. The effects of Lv-RON-siRNA on invasion and drug resistance of HT-29 cells were observed by Transwell assay and ATP-TCA (ATP-tumor chemosensitivity assay). Results: The silencing effect of Lv-RON-siRNA on RON gene expression in HT-29 cells reached 70%. Compared with the control group, the invasion of HT-29 cells in Lv-RON-siRNA infection group was decreased（097±0.07 vs 1.29±0.08, P<0.05). The values of IC90 and IC50 of HT-29 cells infected with Lv-RON-siRNA to 5-FU were (14.28±1.34) μg/ml and (8.93±1.2) μg/ml, respectively. The IC90 and IC50 of HT-29 cells infected with Lv-RON-siRNA to cisplatin (DDP) were (1.91±0.22) μg/ml and (0.64±0.07) μg/ml, respectively, and were significantly lower than those in the control group (P<0.01). Conclusion: Silencing RON gene expression can decrease the invasion ability of HT-29 cells and increase the sensitivity of HT-29 cells to 5-FU and DDP.
8 Effect of triterpenes compound of cortex periplocae on PCNA expression in rat esophageal carcinoma
2012, 19(5):508-512.
DOI: 10.3872/j.issn.1007-385X.2012.5.008
Abstract:
Objective : To investigate the effect of triterpenes compound of cortex periplocae (TCCP) on the expression of proliferating cell nuclear antigen (PCNA) in N-nitrosomethylbenzylamine (NMBA)-induced rat esophageal carcinoma. Methods: 120 male F344 rats were randomly divided into four groups: a model group treated with NMBA only and a TCCP treatment group receiving NMBA plus TCCP, a soya oil control group treated with soya oil and a normal control group. At 9, 15 and 25 weeks after treatment, the pathological changes of esophageal tissues were detected by H-E staining, while the expression of PCNA was measured by immunohistochemistry. Results: There were no abnormal changes in the normal and soya oil groups. At 9, 15 and 25 weeks after treatment, the incidence of preneoplastic lesion in the NMBA group was 20.0%, 46.7% and 933%, respectively. At 9 and 15 weeks, the rate of rat esophageal preneoplastic lesion significantly decreased in TCCP treatment group compared with NMBA group (0,0 vs 20.0%, 46.7%, P<0.05). There was a significant increase in the expression of PCNA in the NMBA group (week 9: 213.17±29.74; week 15: 268.35±39.56; week 25: 327.24±28.19), compared with that in the normal control group (\[167.96±2016\], \[170.76±14.79\], \[172.49±17.49\], P<0.05). TCCP significantly decreased the expression of PCNA compared with that in the NMBA group (P<0.05). Conclusion: TCCP inhibits NMBA-induced rat esophageal carcinoma probably via suppression of PCNA expression.
Objective : To investigate the effect of triterpenes compound of cortex periplocae (TCCP) on the expression of proliferating cell nuclear antigen (PCNA) in N-nitrosomethylbenzylamine (NMBA)-induced rat esophageal carcinoma. Methods: 120 male F344 rats were randomly divided into four groups: a model group treated with NMBA only and a TCCP treatment group receiving NMBA plus TCCP, a soya oil control group treated with soya oil and a normal control group. At 9, 15 and 25 weeks after treatment, the pathological changes of esophageal tissues were detected by H-E staining, while the expression of PCNA was measured by immunohistochemistry. Results: There were no abnormal changes in the normal and soya oil groups. At 9, 15 and 25 weeks after treatment, the incidence of preneoplastic lesion in the NMBA group was 20.0%, 46.7% and 933%, respectively. At 9 and 15 weeks, the rate of rat esophageal preneoplastic lesion significantly decreased in TCCP treatment group compared with NMBA group (0,0 vs 20.0%, 46.7%, P<0.05). There was a significant increase in the expression of PCNA in the NMBA group (week 9: 213.17±29.74; week 15: 268.35±39.56; week 25: 327.24±28.19), compared with that in the normal control group (\[167.96±2016\], \[170.76±14.79\], \[172.49±17.49\], P<0.05). TCCP significantly decreased the expression of PCNA compared with that in the NMBA group (P<0.05). Conclusion: TCCP inhibits NMBA-induced rat esophageal carcinoma probably via suppression of PCNA expression.
ZHANG Hong-li
,
ZENG Peng-yun
,
DENG Li-li
,
ZHANG Lian-sheng
,
CHAI Ye
,
YUE Ling-ling
,
WU Chong-yang
,
LI Li-juan
,
HAO Zheng-dong
,
LI Liang-liang
2012, 19(5):513-516.
DOI: 10.3872/j.issn.1007-385X.2012.5.009
Abstract:
Objective : To investigate the effects of bortezomib on the cytotoxic sensitivity of drug-resistant K562/ADM cells to natural killer (NK) cells and the underlying mechanisms. Methods: The expressions of MICA protein and mRNA on K562/ADM target cells before and after incubation with bortezomib were detected by flow cytometry and real-time PCR, respectively. The cytotoxic sensitivity of K562/ADM cells treated with or without bortezomib to NK cells was measured by LDH releasing assay. Results: The expression rates of MICA protein on K562/ADM cells incubated with bortezomib increased from (17.03±4.94)% to (23.77±5.26)% (P<0.05). The mRNA expression of MICA on K562/ADM cells treated with bortezomib increased (2.03±0.33) times. At the E∶T ratio of 10∶1 and 20∶1, the cytotoxic sensitivity of K562/ADM cells to NK cells increased from (23.22±3.03)% and (30.30±0.74)% in untreated cells to (33.69±128)% and (41.40±1.97)% in bortezomib-treated cells, respectively, showing significant differences (P<0.05). Conclusion: Bortezomib can up-regulate the MICA expression in K562/ADM cells and thus may enhance the cytotoxicity of NK cells against K562/ADM cells.
Objective : To investigate the effects of bortezomib on the cytotoxic sensitivity of drug-resistant K562/ADM cells to natural killer (NK) cells and the underlying mechanisms. Methods: The expressions of MICA protein and mRNA on K562/ADM target cells before and after incubation with bortezomib were detected by flow cytometry and real-time PCR, respectively. The cytotoxic sensitivity of K562/ADM cells treated with or without bortezomib to NK cells was measured by LDH releasing assay. Results: The expression rates of MICA protein on K562/ADM cells incubated with bortezomib increased from (17.03±4.94)% to (23.77±5.26)% (P<0.05). The mRNA expression of MICA on K562/ADM cells treated with bortezomib increased (2.03±0.33) times. At the E∶T ratio of 10∶1 and 20∶1, the cytotoxic sensitivity of K562/ADM cells to NK cells increased from (23.22±3.03)% and (30.30±0.74)% in untreated cells to (33.69±128)% and (41.40±1.97)% in bortezomib-treated cells, respectively, showing significant differences (P<0.05). Conclusion: Bortezomib can up-regulate the MICA expression in K562/ADM cells and thus may enhance the cytotoxicity of NK cells against K562/ADM cells.
2012, 19(5):517-520.
DOI: 10.3872/j.issn.1007-385X.2012.5.010
Abstract:
Objective : To obverse the effect of interleukin-21(IL-21) on CD4+CD25+ regulatory T cells (Tregs) in chronic myeloid leukemia (CML) patients. Methods: Peripheral blood mononuclear cells (PBMCs) were collected from newly diagnosed CML patients in the Hematology Department of Second Hospital of Lanzhou University, and CD4+T cells were isolated by magnetic beads. CD4+T cells were co-cultured with IL-21 (50, 100, and 200 ng/ml) and saline respectively for 72 h, and then the Treg ratio was determined by flow cytometry; Foxp3 mRNA expression was examined by real-time PCR; and IL-10 and TGF-β in different cell supernatants were detected by ELISA. Results: Compared with the control group, the ratios of Treg in CD4+T cells were decreased in 50 ng/ml, 100 ng/ml and 200 ng/ml IL-21 groups (\[342±0.76\]%, \[6.81±0.33\]%, \[7.98±0.76\]% vs \[12.09±0.91\]%， P<0.05); and Foxp3 mRNA expressions were also decreased (\[0.05±0.02\], \[0.16±002\], \[0.25±0.02\] vs 1, P<0.05), the levels of IL-10 (\[2613±7.28\], \[44.88±3.72\], \[79.77±394\] vs \[133.00±12.32\] pg/ml, P<0.05) and TGF-β (\[9.25±0.84\], \[16.70±1.00\], \[20.47±1.59\] vs \[26.05±1.81\] pg/ml, P<0.05) in cell supernatants were significantly decreased. Conclusion: IL-21 can inhibit the ratio and function of Treg in peripheral blood of CML patients.
Objective : To obverse the effect of interleukin-21(IL-21) on CD4+CD25+ regulatory T cells (Tregs) in chronic myeloid leukemia (CML) patients. Methods: Peripheral blood mononuclear cells (PBMCs) were collected from newly diagnosed CML patients in the Hematology Department of Second Hospital of Lanzhou University, and CD4+T cells were isolated by magnetic beads. CD4+T cells were co-cultured with IL-21 (50, 100, and 200 ng/ml) and saline respectively for 72 h, and then the Treg ratio was determined by flow cytometry; Foxp3 mRNA expression was examined by real-time PCR; and IL-10 and TGF-β in different cell supernatants were detected by ELISA. Results: Compared with the control group, the ratios of Treg in CD4+T cells were decreased in 50 ng/ml, 100 ng/ml and 200 ng/ml IL-21 groups (\[342±0.76\]%, \[6.81±0.33\]%, \[7.98±0.76\]% vs \[12.09±0.91\]%， P<0.05); and Foxp3 mRNA expressions were also decreased (\[0.05±0.02\], \[0.16±002\], \[0.25±0.02\] vs 1, P<0.05), the levels of IL-10 (\[2613±7.28\], \[44.88±3.72\], \[79.77±394\] vs \[133.00±12.32\] pg/ml, P<0.05) and TGF-β (\[9.25±0.84\], \[16.70±1.00\], \[20.47±1.59\] vs \[26.05±1.81\] pg/ml, P<0.05) in cell supernatants were significantly decreased. Conclusion: IL-21 can inhibit the ratio and function of Treg in peripheral blood of CML patients.
2012, 19(5):521-525.
DOI: 10.3872/j.issn.1007-385X.2012.5.011
Abstract:
Objective : To detect the expression of γ-synuclein in esophageal adenocarcinoma and evaluate its effect on invasion of esophageal carcinoma Eca-109 cells. Methods: SP immunohistochemical staining was used to examine the expression of γ-synuclein in 30 esophageal carcinoma specimens and 10 normal control specimens from January to December 2010 (Fourth Hospital of Hebei Medical University). The plasmid containing γ-synuclein (pcDNA3.0-γ-synuclein) was transfected into Eca-109 cells by lipofectamine. The expressions of γ-synuclein mRNA and protein were investigated by RT-PCR and flow cytometry, respectively. The invasion ability of transfected Eca-109 cells was dectected by Transwell assay. Results: The positive exppression rate of γ-synuclein was 83% in human esophageal carcinoma, which was significantly higher than that in normal esophageal tissues (P<0.05), and the positive rate of γ-synuclein in stage-Ⅱ esophageal carcinoma patients was significantly lower than that in stage Ⅲ-Ⅳ patients (22.2% vs 76.2%，P<0.05). By comparing transfected group in Eca-109 cells with empty plasmid group and control group, RT-PCR showed the expression level of γ-synuclein mRNA increased significantly (\[1.10±0.03\] vs \[0.42±0.03\]，\[0.45±0.15\]; P<0.01). Flow cytometry showed that the expression level of γ-synuclein protein increased significantly compared with the other two groups (\[1.05±0.06\] vs \[0.80±0.45\], \[0.79±0.46\]; P<0.05). Invasion through matrigel abilities of Eca-109 cells increased significantly (\[167±2.51\] vs \[65±2.60\], \[70±2.50\]; P<0.01). Conclusion: γ-synuclein is highly expressed in esophageal carcinoma tissues and can enhance the invasion of Eca-109 cells, which may play an important role in carcinogenesis progression and metastasis of esophageal carcinoma.
Objective : To detect the expression of γ-synuclein in esophageal adenocarcinoma and evaluate its effect on invasion of esophageal carcinoma Eca-109 cells. Methods: SP immunohistochemical staining was used to examine the expression of γ-synuclein in 30 esophageal carcinoma specimens and 10 normal control specimens from January to December 2010 (Fourth Hospital of Hebei Medical University). The plasmid containing γ-synuclein (pcDNA3.0-γ-synuclein) was transfected into Eca-109 cells by lipofectamine. The expressions of γ-synuclein mRNA and protein were investigated by RT-PCR and flow cytometry, respectively. The invasion ability of transfected Eca-109 cells was dectected by Transwell assay. Results: The positive exppression rate of γ-synuclein was 83% in human esophageal carcinoma, which was significantly higher than that in normal esophageal tissues (P<0.05), and the positive rate of γ-synuclein in stage-Ⅱ esophageal carcinoma patients was significantly lower than that in stage Ⅲ-Ⅳ patients (22.2% vs 76.2%，P<0.05). By comparing transfected group in Eca-109 cells with empty plasmid group and control group, RT-PCR showed the expression level of γ-synuclein mRNA increased significantly (\[1.10±0.03\] vs \[0.42±0.03\]，\[0.45±0.15\]; P<0.01). Flow cytometry showed that the expression level of γ-synuclein protein increased significantly compared with the other two groups (\[1.05±0.06\] vs \[0.80±0.45\], \[0.79±0.46\]; P<0.05). Invasion through matrigel abilities of Eca-109 cells increased significantly (\[167±2.51\] vs \[65±2.60\], \[70±2.50\]; P<0.01). Conclusion: γ-synuclein is highly expressed in esophageal carcinoma tissues and can enhance the invasion of Eca-109 cells, which may play an important role in carcinogenesis progression and metastasis of esophageal carcinoma.
2012, 19(5):526-530.
DOI: 10.3872/j.issn.1007-385X.2012.5.012
Abstract:
Objective : To investigate the methylation of apoptosis protease activating factor-1 ( Apaf-1) gene promoter and its correlation with protein expression of enhancer of zeste homolog 2 (EZH2) in gastric cardia adenocarcinoma (GCA). Methods: Methylation specific polymerase chain reaction (MSP) method was used to examine the methylation status of Apaf-1 gene, and immunohistochemistry method was used to detect the protein expression of Apaf-1 and EZH2 in GCA and paracancerous tissues. Results: The frequency of Apaf-1 methylation in GCA tissues (49.6%, 62/125) was significantly higher than that in paracancerous tissues (4.0%, 5/125, P<0.01). The methylation frequencies of Apaf-1gene in stage Ⅲ and Ⅳ GCA tissues were significantly higher than those in stage Ⅰ and Ⅱ GCA tissues (58.8% vs 38.6%, P<0.05). No significant difference in the methylation frequency of Apaf-1 gene was found in three histological differentiated groups (P＞0.05). The positive protein expression of Apaf-1 in GCA tissues was significantly decreased compared with paracancerous tissues (39.2% vs 96.0%, P<0.01) and was correlated with its methylation status. The positive protein expression of EZH2 in GCA tissues was significantly increased compared with that in paracancerous tissues (744% vs 11.2%, P<0.01) and was negatively correlated with the expression of Apaf-1 protein in GCA. Conclusion: The promoter of Apaf-1 gene is hypermethylated and both Apaf-1 and EZH2 may play important roles in the development of GCA.
Objective : To investigate the methylation of apoptosis protease activating factor-1 ( Apaf-1) gene promoter and its correlation with protein expression of enhancer of zeste homolog 2 (EZH2) in gastric cardia adenocarcinoma (GCA). Methods: Methylation specific polymerase chain reaction (MSP) method was used to examine the methylation status of Apaf-1 gene, and immunohistochemistry method was used to detect the protein expression of Apaf-1 and EZH2 in GCA and paracancerous tissues. Results: The frequency of Apaf-1 methylation in GCA tissues (49.6%, 62/125) was significantly higher than that in paracancerous tissues (4.0%, 5/125, P<0.01). The methylation frequencies of Apaf-1gene in stage Ⅲ and Ⅳ GCA tissues were significantly higher than those in stage Ⅰ and Ⅱ GCA tissues (58.8% vs 38.6%, P<0.05). No significant difference in the methylation frequency of Apaf-1 gene was found in three histological differentiated groups (P＞0.05). The positive protein expression of Apaf-1 in GCA tissues was significantly decreased compared with paracancerous tissues (39.2% vs 96.0%, P<0.01) and was correlated with its methylation status. The positive protein expression of EZH2 in GCA tissues was significantly increased compared with that in paracancerous tissues (744% vs 11.2%, P<0.01) and was negatively correlated with the expression of Apaf-1 protein in GCA. Conclusion: The promoter of Apaf-1 gene is hypermethylated and both Apaf-1 and EZH2 may play important roles in the development of GCA.
2012, 19(5):531-534.
DOI: 10.3872/j.issn.1007-385X.2012.5.013
Abstract:
Objective : To study the clinicopathological significance of the expressions of ERK, C-Jun and JAK2 in hepatic cancer tissues and adjacent tumor tissues. Methods: The expressions of ERK, C-Jun and JAK2 in 60 hepatic cancer specimens and adjacent tumor tissues (from Department of Gastroenterology, Affiliated Hospital of Medical College of Chinese People’s Armed Police Forces) were detected by immunohistochemical SP method. A Cox multivariate analysis was performed in those patients to study the factors that influence the prognosis of hepatic cancer. Results: The average values of ERK, C-Jun and JAK2 in hepatic cancer tissues were significantly higher than that in the tumor-adjacent tissues (\[0.23±0.03\], \[0.18±0.06\], \[0.19±0.07\] vs \[0.16±0.02\], \[0.13±0.02\], \[0.14±0.05\], P<0.01). The univariate analysis results showed that the major significant prognostic factors influencing survival was the expression of C-Jun and JAK2 (P<005), and multivariate analysis revealed that the expression of JAK2 was the most important prognostic factor for hepatic cancer (P<0.05). Conclusion: The expression of JAK2 may be an prognostic factor for hepatic cancer patients.
Objective : To study the clinicopathological significance of the expressions of ERK, C-Jun and JAK2 in hepatic cancer tissues and adjacent tumor tissues. Methods: The expressions of ERK, C-Jun and JAK2 in 60 hepatic cancer specimens and adjacent tumor tissues (from Department of Gastroenterology, Affiliated Hospital of Medical College of Chinese People’s Armed Police Forces) were detected by immunohistochemical SP method. A Cox multivariate analysis was performed in those patients to study the factors that influence the prognosis of hepatic cancer. Results: The average values of ERK, C-Jun and JAK2 in hepatic cancer tissues were significantly higher than that in the tumor-adjacent tissues (\[0.23±0.03\], \[0.18±0.06\], \[0.19±0.07\] vs \[0.16±0.02\], \[0.13±0.02\], \[0.14±0.05\], P<0.01). The univariate analysis results showed that the major significant prognostic factors influencing survival was the expression of C-Jun and JAK2 (P<005), and multivariate analysis revealed that the expression of JAK2 was the most important prognostic factor for hepatic cancer (P<0.05). Conclusion: The expression of JAK2 may be an prognostic factor for hepatic cancer patients.
2012, 19(5):535-538.
DOI: 10.3872/j.issn.1007-385X.2012.5.014
Abstract:
Objective : To evaluate the clinical efficacy of rituximab (R) combined CHOP ( R-CHOP) treatment on B-cell non-Hodgkin＇s lymphoma (NHL) and its influence on the expression of calreticulin (CRT). Methods: Totally 48 patients (From Jul. 2008 to Feb. 2011 in Affiliated Hospital of Nantong University) with previously untreated B-cell lymphoma were divided into R-CHOP group (25 cases) and CHOP group (23 cases). R-CHOP group was treated with R-CHOP chemotherapy and CHOP group was treated with CHOP chemotherapy. After 6 treatment courses, the clinical efficacy, adverse effect and the expression of CRT on CD20+ B cells were compared between these two groups. Results: The complete remission rate was 80.0% and the total effective rate was 92.0% in the R-CHOP group. The complete remission rate was 56.5% and the total effective rate was 696% in the CHOP group. There was a significant difference in the complete remission rate and the total effective rate between the two groups (P<0.05). There was no significant difference in adverse events between the two groups (P>005). The expression of CRT on CD20+ B cells in the R-CHOP group was higher than the CHOP group (\[255.00±5.57\] vs \[216.00±3.61\], P<0.05). Conclusion: Rituximab can improve the clinical effciency of CHOP in the treatment of B-cell NHL, which may be related with the expression of CRT induced by combined treatment.
Objective : To evaluate the clinical efficacy of rituximab (R) combined CHOP ( R-CHOP) treatment on B-cell non-Hodgkin＇s lymphoma (NHL) and its influence on the expression of calreticulin (CRT). Methods: Totally 48 patients (From Jul. 2008 to Feb. 2011 in Affiliated Hospital of Nantong University) with previously untreated B-cell lymphoma were divided into R-CHOP group (25 cases) and CHOP group (23 cases). R-CHOP group was treated with R-CHOP chemotherapy and CHOP group was treated with CHOP chemotherapy. After 6 treatment courses, the clinical efficacy, adverse effect and the expression of CRT on CD20+ B cells were compared between these two groups. Results: The complete remission rate was 80.0% and the total effective rate was 92.0% in the R-CHOP group. The complete remission rate was 56.5% and the total effective rate was 696% in the CHOP group. There was a significant difference in the complete remission rate and the total effective rate between the two groups (P<0.05). There was no significant difference in adverse events between the two groups (P>005). The expression of CRT on CD20+ B cells in the R-CHOP group was higher than the CHOP group (\[255.00±5.57\] vs \[216.00±3.61\], P<0.05). Conclusion: Rituximab can improve the clinical effciency of CHOP in the treatment of B-cell NHL, which may be related with the expression of CRT induced by combined treatment.
2012, 19(5):539-542.
DOI: 10.3872/j.issn.1007-385X.2012.5.015
Abstract:
Objective : To explore the relationship between the expression levels of P110α and pAKT in esophageal carcinoma (EC) tissues with clinical features of EC. Methods: EC tissues from 76 patients and esophageal tissues from 25 normal controls were collected from the Department of Pathology, No. 101 Hospital of PLA during May 2010 to May 2011. The expression levels of P110α and pAKT in EC and normal esophageal tissues were detected by SP immunohistochemistry. Results: The positive rates of pAKT and P110α in the EC group were 72.4% (55/76) and 57.9% (44/76), respectively, which were significantly higher than those in the control group（PpAKT=0.001，PP110α=0025）. The high level of protein expressions of pAKT and P110α were correlated with lymph node metastasis of EC（PpAKT=0.017，PP110α=0009). However, no correlation was found with other clinical pathological features. The double positive rate of pAKT and P110α expression in EC was 52.6% (40/76). The expression of pAKT was found in a positive correlation with P110α(r=0.486，P=0.001). Conclusion: P110α and pAKT expressions in EC are correlated with lymph node metastasis, and P110α and pAKT might be used as markers of lymph node metastasis in EC.
Objective : To explore the relationship between the expression levels of P110α and pAKT in esophageal carcinoma (EC) tissues with clinical features of EC. Methods: EC tissues from 76 patients and esophageal tissues from 25 normal controls were collected from the Department of Pathology, No. 101 Hospital of PLA during May 2010 to May 2011. The expression levels of P110α and pAKT in EC and normal esophageal tissues were detected by SP immunohistochemistry. Results: The positive rates of pAKT and P110α in the EC group were 72.4% (55/76) and 57.9% (44/76), respectively, which were significantly higher than those in the control group（PpAKT=0.001，PP110α=0025）. The high level of protein expressions of pAKT and P110α were correlated with lymph node metastasis of EC（PpAKT=0.017，PP110α=0009). However, no correlation was found with other clinical pathological features. The double positive rate of pAKT and P110α expression in EC was 52.6% (40/76). The expression of pAKT was found in a positive correlation with P110α(r=0.486，P=0.001). Conclusion: P110α and pAKT expressions in EC are correlated with lymph node metastasis, and P110α and pAKT might be used as markers of lymph node metastasis in EC.
2012, 19(5):543-546.
DOI: 10.3872/j.issn.1007-385X.2012.5.016
Abstract:
目的：探讨贝伐单抗（bevacizumab）对人非小细胞肺癌A549-luc细胞小鼠移植瘤生长、浸润的影响。 方法： 将稳定表达荧光素酶的人肺癌细胞系A549-Luc接种于裸鼠皮下，成瘤后随机分为对照组、低剂量贝伐单抗组及高剂量贝伐单抗组。对移植瘤进行生物发光成像，并与体积测量结果拟合；检测移植瘤质量和体积，计算贝伐单抗的抑瘤率。 结果： 生物发光成像观察结果显示，贝伐单抗组移植瘤发光区域和荧光强度较对照组均显著降低，且荧光高峰值出现较晚，高剂量贝伐单抗组的抑制效果更为显著。随着治疗时间的延长，贝伐单抗组移植瘤体积增长速度小于对照组（P<0.05），高剂量贝伐单抗组移植瘤体积缩小更为显著（P<0.05）。贝伐单抗组移植瘤细胞浸润程度和对肺组织的破坏程度均低于对照组，高剂量贝伐单抗组抑瘤率为77.3%，高于低剂量组的52.3%（P<0.05）。 结论： 贝伐单抗可以有效抑制活体内非小细胞肺癌A549-luc细胞移植瘤的生长和浸润。
目的：探讨贝伐单抗（bevacizumab）对人非小细胞肺癌A549-luc细胞小鼠移植瘤生长、浸润的影响。 方法： 将稳定表达荧光素酶的人肺癌细胞系A549-Luc接种于裸鼠皮下，成瘤后随机分为对照组、低剂量贝伐单抗组及高剂量贝伐单抗组。对移植瘤进行生物发光成像，并与体积测量结果拟合；检测移植瘤质量和体积，计算贝伐单抗的抑瘤率。 结果： 生物发光成像观察结果显示，贝伐单抗组移植瘤发光区域和荧光强度较对照组均显著降低，且荧光高峰值出现较晚，高剂量贝伐单抗组的抑制效果更为显著。随着治疗时间的延长，贝伐单抗组移植瘤体积增长速度小于对照组（P<0.05），高剂量贝伐单抗组移植瘤体积缩小更为显著（P<0.05）。贝伐单抗组移植瘤细胞浸润程度和对肺组织的破坏程度均低于对照组，高剂量贝伐单抗组抑瘤率为77.3%，高于低剂量组的52.3%（P<0.05）。 结论： 贝伐单抗可以有效抑制活体内非小细胞肺癌A549-luc细胞移植瘤的生长和浸润。
2012, 19(5):547-549.
DOI: 10.3872/j.issn.1007-385X.2012.5.017
Abstract:
目的：探讨乏氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）在乏氧状态下对鼻咽癌CNE-1细胞放射敏感性的影响。 方法： 氯化钴处理CNE-1细胞模拟乏氧条件下的培养传代，通过脂质体将不同 HIF-1α反义寡核苷酸酸导入CNE-1细胞中，根据转染复合物的不同将CNE-1细胞分为反义联合组、正义联合组、脂质体组和单纯放疗组（放疗对照组）。X射线单次照射，照射条件为照射率4 Gy/min，共8 Gy，细胞照射后继续乏氧条件下培养24 h。MTT法检测CNE-1细胞的存活率，Western blotting检测CNE-1细胞中HIF-1α和VEGF蛋白的表达。 结果： 在乏氧条件下，反义联合组的鼻咽癌CNE-1细胞存活率明显大于正义联合组和单纯放疗组。Western blotting结果显示，与正义联合组和单纯放疗组相比，反义联合组CNE-1细胞中HIF-1α蛋白表达显著下降 （0.162±0.055 vs 0.872±0.191，0.768±0.217，0.863±0.245，P<0.05）；同时反义联合组CNE-1细胞中VEGF蛋白的表达量较正义联合组和单纯放疗组明显减少 （0.364±0.078 vs 1.165±0.346，1.068±0379，1087±0.266，P<0.05）。 结论： HIF-1α反义寡核苷酸能有效抑制HIF-1α的表达，对乏氧状态下的鼻咽癌CNE-1细胞具有放射增敏作用。
目的：探讨乏氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）在乏氧状态下对鼻咽癌CNE-1细胞放射敏感性的影响。 方法： 氯化钴处理CNE-1细胞模拟乏氧条件下的培养传代，通过脂质体将不同 HIF-1α反义寡核苷酸酸导入CNE-1细胞中，根据转染复合物的不同将CNE-1细胞分为反义联合组、正义联合组、脂质体组和单纯放疗组（放疗对照组）。X射线单次照射，照射条件为照射率4 Gy/min，共8 Gy，细胞照射后继续乏氧条件下培养24 h。MTT法检测CNE-1细胞的存活率，Western blotting检测CNE-1细胞中HIF-1α和VEGF蛋白的表达。 结果： 在乏氧条件下，反义联合组的鼻咽癌CNE-1细胞存活率明显大于正义联合组和单纯放疗组。Western blotting结果显示，与正义联合组和单纯放疗组相比，反义联合组CNE-1细胞中HIF-1α蛋白表达显著下降 （0.162±0.055 vs 0.872±0.191，0.768±0.217，0.863±0.245，P<0.05）；同时反义联合组CNE-1细胞中VEGF蛋白的表达量较正义联合组和单纯放疗组明显减少 （0.364±0.078 vs 1.165±0.346，1.068±0379，1087±0.266，P<0.05）。 结论： HIF-1α反义寡核苷酸能有效抑制HIF-1α的表达，对乏氧状态下的鼻咽癌CNE-1细胞具有放射增敏作用。
2012, 19(5):550-555.
DOI: 10.3872/j.issn.1007-385X.2012.5.018
Abstract:
骨髓增生异常综合征（myelodysplastic syndrome，MDS）的发病机制涉及多阶段、多因素，基因改变与表观遗传修饰可能共同参与了这一过程。DNA甲基化是表观遗传学中一种最为重要的修饰，MDS患者常表现为总体DNA高甲基化。使用DNA甲基转移酶（DNA methyltransferase，DNMT）抑制剂降低总体甲基化水平，在MDS患者中取得了富有成效的临床反应及血液学改善。DNMT抑制剂可分为两类：5-氮杂胞苷(5-azacytidine, 5-Aza-CdR)、地西他滨（5-Aza-2-deoxycytidine, decitabine）等核苷和核苷衍生物类抑制剂，它们可提高MDS患者的临床完全反应率、部分反应率及血液学改善，但缓解率、疗效尚不够令人满意；肼苯哒嗪等非核苷类抑制剂。非核苷类抑制剂与丙戊酸镁联合应用治疗MDS获得成功，为MDS去甲基化治疗药物的研究开启了一种新思路。
骨髓增生异常综合征（myelodysplastic syndrome，MDS）的发病机制涉及多阶段、多因素，基因改变与表观遗传修饰可能共同参与了这一过程。DNA甲基化是表观遗传学中一种最为重要的修饰，MDS患者常表现为总体DNA高甲基化。使用DNA甲基转移酶（DNA methyltransferase，DNMT）抑制剂降低总体甲基化水平，在MDS患者中取得了富有成效的临床反应及血液学改善。DNMT抑制剂可分为两类：5-氮杂胞苷(5-azacytidine, 5-Aza-CdR)、地西他滨（5-Aza-2-deoxycytidine, decitabine）等核苷和核苷衍生物类抑制剂，它们可提高MDS患者的临床完全反应率、部分反应率及血液学改善，但缓解率、疗效尚不够令人满意；肼苯哒嗪等非核苷类抑制剂。非核苷类抑制剂与丙戊酸镁联合应用治疗MDS获得成功，为MDS去甲基化治疗药物的研究开启了一种新思路。
2012, 19(5):556-560.
DOI: 10.3872/j.issn.1007-385X.2012.5.019
Abstract:
膜联蛋白A2（annexin A2，AXⅡ）是依赖钙离子调节的膜磷脂结合的蛋白质多基因家族中的一员，具有多种生物学功能，包括参与细胞增殖、凋亡、信号转导、细胞迁移、DNA的合成复制、RNA结合等。AXⅡ在肝癌、胰腺癌、骨肉瘤等组织尤其是消化系统肿瘤中高表达，而在食管癌、前列腺癌组织中表达下调，说明AXⅡ在不同肿瘤发生、发展中起不同作用。AXⅡ在肿瘤细胞中主要发挥分子调节作用，如血管生成、增殖、凋亡、细胞侵袭、转移和黏附，因此，AXⅡ与肿瘤的临床诊断分期与预后、治疗密切相关。
膜联蛋白A2（annexin A2，AXⅡ）是依赖钙离子调节的膜磷脂结合的蛋白质多基因家族中的一员，具有多种生物学功能，包括参与细胞增殖、凋亡、信号转导、细胞迁移、DNA的合成复制、RNA结合等。AXⅡ在肝癌、胰腺癌、骨肉瘤等组织尤其是消化系统肿瘤中高表达，而在食管癌、前列腺癌组织中表达下调，说明AXⅡ在不同肿瘤发生、发展中起不同作用。AXⅡ在肿瘤细胞中主要发挥分子调节作用，如血管生成、增殖、凋亡、细胞侵袭、转移和黏附，因此，AXⅡ与肿瘤的临床诊断分期与预后、治疗密切相关。
2012, 19(5):561-564.
DOI: 10.3872/j.issn.1007-385X.2012.5.020
Abstract:
Twist属于高度保守的碱性螺旋-环-螺旋（basichelix-loop-helix，bHLH）结构的转录因子家族，在动物和人的胚胎发育、诱导细胞迁移以及组织塑形中起重要作用。Twist基因具有癌基因特征，在多种恶性肿瘤中高表达，不仅参与肿瘤的发生、侵袭和转移，而且与肿瘤细胞产生耐药性密切相关。Twist基因可以通过抗细胞凋亡、干扰抗微管类药物、诱发上皮-间质转型、诱导产生肿瘤干细胞样特性及影响肿瘤微环境等促使肿瘤细胞耐药，因此有望成为逆转肿瘤耐药的新靶点。
Twist属于高度保守的碱性螺旋-环-螺旋（basichelix-loop-helix，bHLH）结构的转录因子家族，在动物和人的胚胎发育、诱导细胞迁移以及组织塑形中起重要作用。Twist基因具有癌基因特征，在多种恶性肿瘤中高表达，不仅参与肿瘤的发生、侵袭和转移，而且与肿瘤细胞产生耐药性密切相关。Twist基因可以通过抗细胞凋亡、干扰抗微管类药物、诱发上皮-间质转型、诱导产生肿瘤干细胞样特性及影响肿瘤微环境等促使肿瘤细胞耐药，因此有望成为逆转肿瘤耐药的新靶点。
2012, 19(5):565-568.
DOI: 10.3872/j.issn.1007-385X.2012.5.021
Abstract:
腺相关病毒（adeno-associated virus，AAV）是一类单链线状DNA缺陷性病毒，属细小病毒科，是基因转移最为理想的载体之一。AAV介导肿瘤相关基因感染树突状细胞（dendritic cell，DC）诱导细胞毒性T淋巴细胞（cytotoxic T lymphocyte，CTL），可以内源性持续表达肿瘤相关抗原，并由MHCⅠ类途径得到充分提呈、放大免疫激发CTL的能力，但不影响DC的成熟度。AAV介导抗肿瘤血管生成基因，有抑制肿瘤生长的作用，并且克服了单克隆抗体生产过程中提纯的困难；AAV介导免疫相关基因具有直接抑制肿瘤细胞增殖或免疫调节的作用；AAV介导抑癌相关基因可以诱导肿瘤细胞凋亡；AAV介导自杀基因可以抑制肿瘤的生长。
腺相关病毒（adeno-associated virus，AAV）是一类单链线状DNA缺陷性病毒，属细小病毒科，是基因转移最为理想的载体之一。AAV介导肿瘤相关基因感染树突状细胞（dendritic cell，DC）诱导细胞毒性T淋巴细胞（cytotoxic T lymphocyte，CTL），可以内源性持续表达肿瘤相关抗原，并由MHCⅠ类途径得到充分提呈、放大免疫激发CTL的能力，但不影响DC的成熟度。AAV介导抗肿瘤血管生成基因，有抑制肿瘤生长的作用，并且克服了单克隆抗体生产过程中提纯的困难；AAV介导免疫相关基因具有直接抑制肿瘤细胞增殖或免疫调节的作用；AAV介导抑癌相关基因可以诱导肿瘤细胞凋亡；AAV介导自杀基因可以抑制肿瘤的生长。
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.