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Volume 24,Issue 12,2017 Table of Contents
2017, 24(12):1343-1349.
DOI: 10.3872/j.issn.1007-385X.2017.12.001
Abstract:
Dynamic change and spatial characteristics of immune response is one of the fundamental theories of precision medical treatment. One certain immune regulator may have totally different functions under different temporal and spatial distribution. T cell immunoglobulin domain and mucin domain protein-3 (TIM-3) is a recently identified immune checkpoint inhibitor. Like PD-1 and CTLA-4, TIM-3 contributes to immune homeostasis. Recently, TIM-3 has attracted much attention as its dysregulated over-expression was approved to be associated with tumors and chronic viral infections.Strategies targeting immune checkpoints are now showing therapeutic prospect. However, the low efficiency rate, therapy resistance and even side effects still exist, which call for precise mechanisms investigation. Most of the research indicates the immune inhibitory effect of TIM-3; still, there are some reports on opposite conclusion. TIM-3 has multiple ligands,and the intra-cellular signaling mechanism is still unclear. In addition, the function and clinical significance of soluble TIM-3 protein in plasma remain unclear. If TIM-3 acquired multiple functions following the evolution of immune system,clarify the mechanisms by which TIM-3 regulates immune response will provided much important information for its further investigation.
Dynamic change and spatial characteristics of immune response is one of the fundamental theories of precision medical treatment. One certain immune regulator may have totally different functions under different temporal and spatial distribution. T cell immunoglobulin domain and mucin domain protein-3 (TIM-3) is a recently identified immune checkpoint inhibitor. Like PD-1 and CTLA-4, TIM-3 contributes to immune homeostasis. Recently, TIM-3 has attracted much attention as its dysregulated over-expression was approved to be associated with tumors and chronic viral infections.Strategies targeting immune checkpoints are now showing therapeutic prospect. However, the low efficiency rate, therapy resistance and even side effects still exist, which call for precise mechanisms investigation. Most of the research indicates the immune inhibitory effect of TIM-3; still, there are some reports on opposite conclusion. TIM-3 has multiple ligands,and the intra-cellular signaling mechanism is still unclear. In addition, the function and clinical significance of soluble TIM-3 protein in plasma remain unclear. If TIM-3 acquired multiple functions following the evolution of immune system,clarify the mechanisms by which TIM-3 regulates immune response will provided much important information for its further investigation.
2017, 24(12):1350-1355.
DOI: 10.3872/j.issn.1007-385X.2017.12.002
Abstract:
The application of monoclonal antibodies to block immune checkpoints including PD-1, CTLA-4 has demonstrated a good efficacy in cancer immunotherapy. However, some patients poorly responded to these immune checkpoints blockers. One probable mechanism for this failure in igniting the antitumor effect was that other inhibitory molecules over expressed in these cases. As an immune checkpoint, T cell immunolobulin and mucin-containing protein-3(TIM-3) is widely expressed in a variety of immune cells, and palys an important role in the regulation of immune response. Many studies showed that lymphocytes from patient peripheral blood or tumor-infiltrating lymphocytes expressed high levels of TIM-3, which was associated with poor outcome. T cell, DC cell and mononuclear phagocytes with over-expressed TIM-3 showed significant inhibitory effects on antitumor immune response. In preclinical studies, combined blockade of TIM-3 and PD-1 with antibodies showed a synergistic effect on antitumor immunity. Clinical trials are underway to evaluate the safety and efficacy of TIM-3 monoclonal antibodies in cancer patients. However, the regulatory role of TIM-3 on immune cells has not been fully clarified, better understanding of the immune modulatory mechanism of TIM-3 helps to develop effective treatment strategy of blockade of TIM-3 in future clinical trials.
The application of monoclonal antibodies to block immune checkpoints including PD-1, CTLA-4 has demonstrated a good efficacy in cancer immunotherapy. However, some patients poorly responded to these immune checkpoints blockers. One probable mechanism for this failure in igniting the antitumor effect was that other inhibitory molecules over expressed in these cases. As an immune checkpoint, T cell immunolobulin and mucin-containing protein-3(TIM-3) is widely expressed in a variety of immune cells, and palys an important role in the regulation of immune response. Many studies showed that lymphocytes from patient peripheral blood or tumor-infiltrating lymphocytes expressed high levels of TIM-3, which was associated with poor outcome. T cell, DC cell and mononuclear phagocytes with over-expressed TIM-3 showed significant inhibitory effects on antitumor immune response. In preclinical studies, combined blockade of TIM-3 and PD-1 with antibodies showed a synergistic effect on antitumor immunity. Clinical trials are underway to evaluate the safety and efficacy of TIM-3 monoclonal antibodies in cancer patients. However, the regulatory role of TIM-3 on immune cells has not been fully clarified, better understanding of the immune modulatory mechanism of TIM-3 helps to develop effective treatment strategy of blockade of TIM-3 in future clinical trials.
3 Inhibitory effect of apopotin-loaded oncolytic adenovirus ATV on human cervical carcinoma HeLa cells
YIN Xunzhe
,
CHEN Shuang
,
LI Wenjie
,
ZHU Yilong
,
LI Yiquan
,
CUI Chuanxin
,
LI Min
,
CUI Yingli
,
ZHAO Jin
,
LI Shanzhi
,
ZHENG Ying
,
SUN Lili
,
LI Xiao
,
GUO Yan
,
JIN Ningyi
2017, 24(12):1356-1361.
DOI: 10.3872/j.issn.1007-385X.2017.12.003
Abstract:
Objective: To investigate the effect of apoptin-loaded oncolytic adenovirus ATV infection on the autophagy and apoptosis of human cervical carcinoma HeLa cells. Methods: Apoptin- loaded oncolytic adenovirus ATV and the control virus Ad-MOCK (both were constructed previously) were transfected into HeLa cells. The effect of ATV infection on proliferation of HeLa cells was measured through WST-1 assay; The effect of ATV infection on apoptosis and cell cycle of HeLa cells was detected by flow cytometry; Western blotting was used to detect the effect of ATV infection on the expression of autophagy-related proteins (LC3, P62 and mTOR) in HeLa cells;MDC staining was used to detect the effect of ATV infection on autophagy of HeLa cells. Results: ATV infection inhibited HeLa cell proliferation in a time-dependent manner; and the inhibitory rate reached 59.26% with ATV 100 MOI at 72 h, which was significantly higher than that of the control group (P<0.01); The apoptotic rate of HeLa cells in ATV group was significantly higher than that in control group[ （38.995±4.009）% vs（14.680±1.174）%,P<0.01] 48 h after infection. With the infection of ATV, flow cytometry analysis showed that the cell cycle of HeLa was blocked at S phase, which was most significant at 48 h and significantly higher than that in control group [ （58.490±2.447)% vs (43.235±4.419)%, P<0.05]. Western blotting revealed that after ATV infection, LC3 expression was gradually increase at 6, 12 h, with a highest level at 12 h（0.368±0.010，P<0.01）and a lowest level at 24 h (0.106 ± 0.023，P<0.01),P62 protein showed the highest expression level at 24 h（6.004 ± 1.423，P<0.01）, while mTOR gradually reduced（0.042±0.010，48 h，P<0.01）with the extension of infection time; under fluorescent microscope,monodansylcadaverin (MDC) positive staining of autophagosomes was observed in the peripheral region of the nucleus of HeLa cells; the number of autophagosomes of ATV group was significantly increased as the ATV infection time prolonged; compared with control group, the number of autophagosomes of ATV group significantly increased at 6 h and 12 h [ (28.000±2.828) vs (8.500±2.121), (37.000±4.243) vs (14.000±1.414), P<0.01], but relatively reduced at 24 h [ (12.000±2.828) vs (17.000±1.414), P<0.01]. Conclusion: Apoptin-loaded oncolytic adenovirus ATV can promote the autophagy and apoptosis of human cervical cancer HeLa cells and then specifically kill tumor cells.
Objective: To investigate the effect of apoptin-loaded oncolytic adenovirus ATV infection on the autophagy and apoptosis of human cervical carcinoma HeLa cells. Methods: Apoptin- loaded oncolytic adenovirus ATV and the control virus Ad-MOCK (both were constructed previously) were transfected into HeLa cells. The effect of ATV infection on proliferation of HeLa cells was measured through WST-1 assay; The effect of ATV infection on apoptosis and cell cycle of HeLa cells was detected by flow cytometry; Western blotting was used to detect the effect of ATV infection on the expression of autophagy-related proteins (LC3, P62 and mTOR) in HeLa cells;MDC staining was used to detect the effect of ATV infection on autophagy of HeLa cells. Results: ATV infection inhibited HeLa cell proliferation in a time-dependent manner; and the inhibitory rate reached 59.26% with ATV 100 MOI at 72 h, which was significantly higher than that of the control group (P<0.01); The apoptotic rate of HeLa cells in ATV group was significantly higher than that in control group[ （38.995±4.009）% vs（14.680±1.174）%,P<0.01] 48 h after infection. With the infection of ATV, flow cytometry analysis showed that the cell cycle of HeLa was blocked at S phase, which was most significant at 48 h and significantly higher than that in control group [ （58.490±2.447)% vs (43.235±4.419)%, P<0.05]. Western blotting revealed that after ATV infection, LC3 expression was gradually increase at 6, 12 h, with a highest level at 12 h（0.368±0.010，P<0.01）and a lowest level at 24 h (0.106 ± 0.023，P<0.01),P62 protein showed the highest expression level at 24 h（6.004 ± 1.423，P<0.01）, while mTOR gradually reduced（0.042±0.010，48 h，P<0.01）with the extension of infection time; under fluorescent microscope,monodansylcadaverin (MDC) positive staining of autophagosomes was observed in the peripheral region of the nucleus of HeLa cells; the number of autophagosomes of ATV group was significantly increased as the ATV infection time prolonged; compared with control group, the number of autophagosomes of ATV group significantly increased at 6 h and 12 h [ (28.000±2.828) vs (8.500±2.121), (37.000±4.243) vs (14.000±1.414), P<0.01], but relatively reduced at 24 h [ (12.000±2.828) vs (17.000±1.414), P<0.01]. Conclusion: Apoptin-loaded oncolytic adenovirus ATV can promote the autophagy and apoptosis of human cervical cancer HeLa cells and then specifically kill tumor cells.
2017, 24(12):1362-1369.
DOI: 10.3872/j.issn.1007-385X.2017.12.004
Abstract:
Objective: To investigate the in vitro synthesis of sgRNA that specific editing CTLA4 (cytotoxic T-lymphocyte-associated protein 4) gene based on the principle of CRISPR/CAS9 gene editing technology. Methods:Firstly, we predicted sgRNAs for CTLA4 locus by CRISPR website, and three sgRNAs（sgRNA1, sgRNA2,sgRNA3）were designed. The recombinant sgRNA plasmids were constructed and transfected into 293T cells. After 48h, the genomic DNA of 293T cells wa s extracted, and the sgRNA sequence with high activity was screened with T7EN1 endonuclease. Subsequently, the screened sgRNA recombinant plasmid was selected as template to design the premier and further amplify the sgRNA core fragment using PCR technology; and then it was transcribed into sgRNA in vitro after optimizing the transcription conditions. Moreover, the cutting efficiency of purified sgRNA was assessed in vitro by Cas9 nuclease. And finally the sgRNA and the transcribed Cas9 mRNA were electroporated into CD3+ T cell of the lung cancer patients to knockout the target gene. Results：The T7EN1 endonuclease experi-ment verified that the knockout rate of sgRNA2 was the maximum with an efficiency up to 66%; after optimization of in vitro synthesis system, the activity and the yield of obtained CTLA4 sgRNA2 were high; it successfully cut the target double- stranded DNA at the designed site, and Cas9 nuclease detected the cutting efficiency of CTLA4 sgRNA2 reached 65%. Finally the CTLA4 gene can be effectively knocked out in CD3+ T cell of the lung cancer patients,and the amount of CTLA4 expression decreased from 46% to 22%, the T7E1 endonuclease experiment also verified that the editing efficiency of sgRNA2 was 56%. Conclusion: A sgRNA with high editing efficiency targeting CTLA4 locus was obtained, which lays a foundation for further establish corresponding immunotherapy strategy targeting CTLA4 gene.
Objective: To investigate the in vitro synthesis of sgRNA that specific editing CTLA4 (cytotoxic T-lymphocyte-associated protein 4) gene based on the principle of CRISPR/CAS9 gene editing technology. Methods:Firstly, we predicted sgRNAs for CTLA4 locus by CRISPR website, and three sgRNAs（sgRNA1, sgRNA2,sgRNA3）were designed. The recombinant sgRNA plasmids were constructed and transfected into 293T cells. After 48h, the genomic DNA of 293T cells wa s extracted, and the sgRNA sequence with high activity was screened with T7EN1 endonuclease. Subsequently, the screened sgRNA recombinant plasmid was selected as template to design the premier and further amplify the sgRNA core fragment using PCR technology; and then it was transcribed into sgRNA in vitro after optimizing the transcription conditions. Moreover, the cutting efficiency of purified sgRNA was assessed in vitro by Cas9 nuclease. And finally the sgRNA and the transcribed Cas9 mRNA were electroporated into CD3+ T cell of the lung cancer patients to knockout the target gene. Results：The T7EN1 endonuclease experi-ment verified that the knockout rate of sgRNA2 was the maximum with an efficiency up to 66%; after optimization of in vitro synthesis system, the activity and the yield of obtained CTLA4 sgRNA2 were high; it successfully cut the target double- stranded DNA at the designed site, and Cas9 nuclease detected the cutting efficiency of CTLA4 sgRNA2 reached 65%. Finally the CTLA4 gene can be effectively knocked out in CD3+ T cell of the lung cancer patients,and the amount of CTLA4 expression decreased from 46% to 22%, the T7E1 endonuclease experiment also verified that the editing efficiency of sgRNA2 was 56%. Conclusion: A sgRNA with high editing efficiency targeting CTLA4 locus was obtained, which lays a foundation for further establish corresponding immunotherapy strategy targeting CTLA4 gene.
2017, 24(12):1370-1374.
DOI: 10.3872/j.issn.1007-385X.2017.12.005
Abstract:
Objective: To investigate the antitumor effect of NRP-1 monoclonal antibody combined with docetaxel metronomic chemotherapy in nude mice bearing gastric cancer xenografts. Methods: BALB / c mice were inoculated subcutaneously with BGC-823 cells to establish xenograft model; the tumor bearing rats were randomly divided into control group, NRP-1 monoclonal antibody group (NRP-1mAb), rhythm chemotherapy group (MCT) and combined group (NRP-1mAb+MCT). Except the control group, the other three groups were given the corresponding treatment on the 8th day after the model establishment. After 2 weeks of administration, the general condition of nude mice was observed and the body weight and tumor volume were measured the next day. The mice were sacrificed and the mass of tumor was calculated; HE staining was used to observe the morphology of the tumor tissue.The expression of NRP-1 protein, VEGF and MVD were detected by immunohistochemistry. Results: The tumor volume (0.613±0.223 vs 0.866±0.115, 1.098±0.343, 1.474±0.644 V/cm3; P<0.05) and weight (0.394±0.128 vs 0.748±0.152, 0.867±0.361, 1.247±0.494 m/g; P<0.05) of the combined group were significantly lower than those of the other groups, and the tumor inhibition rate was statistically higher compared with the other treatment groups (P<0.05). Under microscope, the cancer cells in the control group were well grown and the blood vessels were rich; the cancerous tissues of each treatment group showed different degree of sheet necrosis and the blood vessel composition decreased. Immunohistochemistry results showed that the expression of NRP-1 in the control group was signifi-cantly higher than that in each treatment group (P<0.05); and the expression of NRP-1, VEGF and MVD in combined treatment group was significantly lower than the other groups. Conclusion: NRP-1 monoclonal antibody combined with docetaxel rhythmic chemotherapy may significantly inhibit the growth and angiogenesis of BGC-823 gastric cancer by down-regulating the expression of NRP-1.
Objective: To investigate the antitumor effect of NRP-1 monoclonal antibody combined with docetaxel metronomic chemotherapy in nude mice bearing gastric cancer xenografts. Methods: BALB / c mice were inoculated subcutaneously with BGC-823 cells to establish xenograft model; the tumor bearing rats were randomly divided into control group, NRP-1 monoclonal antibody group (NRP-1mAb), rhythm chemotherapy group (MCT) and combined group (NRP-1mAb+MCT). Except the control group, the other three groups were given the corresponding treatment on the 8th day after the model establishment. After 2 weeks of administration, the general condition of nude mice was observed and the body weight and tumor volume were measured the next day. The mice were sacrificed and the mass of tumor was calculated; HE staining was used to observe the morphology of the tumor tissue.The expression of NRP-1 protein, VEGF and MVD were detected by immunohistochemistry. Results: The tumor volume (0.613±0.223 vs 0.866±0.115, 1.098±0.343, 1.474±0.644 V/cm3; P<0.05) and weight (0.394±0.128 vs 0.748±0.152, 0.867±0.361, 1.247±0.494 m/g; P<0.05) of the combined group were significantly lower than those of the other groups, and the tumor inhibition rate was statistically higher compared with the other treatment groups (P<0.05). Under microscope, the cancer cells in the control group were well grown and the blood vessels were rich; the cancerous tissues of each treatment group showed different degree of sheet necrosis and the blood vessel composition decreased. Immunohistochemistry results showed that the expression of NRP-1 in the control group was signifi-cantly higher than that in each treatment group (P<0.05); and the expression of NRP-1, VEGF and MVD in combined treatment group was significantly lower than the other groups. Conclusion: NRP-1 monoclonal antibody combined with docetaxel rhythmic chemotherapy may significantly inhibit the growth and angiogenesis of BGC-823 gastric cancer by down-regulating the expression of NRP-1.
2017, 24(12):1375-1380.
DOI: 10.3872/j.issn.1007-385X.2017.12.006
Abstract:
Objective: To investigate the effect of Livin gene (an inhibitor of apoptosis protein) on reversing the drug-resistance of osteosarcoma. Methods: Drug-resistant MG-63 cell strain was established in vitro by continuous exposure to doxorubicin at gradually increased concentrations. The resistance index of drug-resistant MG-63 cells was examined by MTT method; Livin protein expressions in MG-63 cells and durg-resistant MG-63 cells were determined by Western blotting．Livin shRNA eukaryotic expression vector (pSilencer3.1-H1 neo-Livin si) was constructed and then transfected into drug-resistant MG-63 cells by using Lipofectmine 2000. Expression change of Livin mRNA and protein in drug-resistant MG-63 cells before and after the transfection was respectively measured by Real-time PCR and Western blotting. The distribution of cell cycle and apoptosis were determined by flow cytometry.The analysis of chemotherapeutic sensitivity of drug-resistant MG-63 cell to doxorubicin was performed by MTT. Results: The recombinant eukaryotic expression vector Silencer3.1-H1 neo-Livin si was successfully constructed.Resistance index to doxorubicin of drug-resistant MG-63 cells (MG-63/R) was 81.32±5.33. Livin shRNA could significantly inhibit the mRNA and protein expression of Livin in MG-63/R cells compared with untransfect-ed group and non-specific transfected group（down-regulated by 72% and 69% at mRNA and protein level respectively,all P<0.05）. The flow cytometry analysis showed there was significantly higher apoptosis rate in Livin shRNA transfected group than that of untransfected group and non- specific transfected group ([22.4±3.2]% vs [4.2±1.1]%, [4.7±0.6]%，P<0.05). After adding doxorubicin, the proliferation of three groups of cells was all inhibited at different levels in time-dependent manner; However, the cell survival rate in Livin shRNA transfected group was significantly lower than that in other two groups (P<0.05). Conclusion：Livin shRNA could efficiently promote the ,apoptosis of drug-resistant MG-63 cells, and thus increase its sensitivity to chemotherapy drugs.
Objective: To investigate the effect of Livin gene (an inhibitor of apoptosis protein) on reversing the drug-resistance of osteosarcoma. Methods: Drug-resistant MG-63 cell strain was established in vitro by continuous exposure to doxorubicin at gradually increased concentrations. The resistance index of drug-resistant MG-63 cells was examined by MTT method; Livin protein expressions in MG-63 cells and durg-resistant MG-63 cells were determined by Western blotting．Livin shRNA eukaryotic expression vector (pSilencer3.1-H1 neo-Livin si) was constructed and then transfected into drug-resistant MG-63 cells by using Lipofectmine 2000. Expression change of Livin mRNA and protein in drug-resistant MG-63 cells before and after the transfection was respectively measured by Real-time PCR and Western blotting. The distribution of cell cycle and apoptosis were determined by flow cytometry.The analysis of chemotherapeutic sensitivity of drug-resistant MG-63 cell to doxorubicin was performed by MTT. Results: The recombinant eukaryotic expression vector Silencer3.1-H1 neo-Livin si was successfully constructed.Resistance index to doxorubicin of drug-resistant MG-63 cells (MG-63/R) was 81.32±5.33. Livin shRNA could significantly inhibit the mRNA and protein expression of Livin in MG-63/R cells compared with untransfect-ed group and non-specific transfected group（down-regulated by 72% and 69% at mRNA and protein level respectively,all P<0.05）. The flow cytometry analysis showed there was significantly higher apoptosis rate in Livin shRNA transfected group than that of untransfected group and non- specific transfected group ([22.4±3.2]% vs [4.2±1.1]%, [4.7±0.6]%，P<0.05). After adding doxorubicin, the proliferation of three groups of cells was all inhibited at different levels in time-dependent manner; However, the cell survival rate in Livin shRNA transfected group was significantly lower than that in other two groups (P<0.05). Conclusion：Livin shRNA could efficiently promote the ,apoptosis of drug-resistant MG-63 cells, and thus increase its sensitivity to chemotherapy drugs.
2017, 24(12):1381-1385.
DOI: 10.3872/j.issn.1007-385X.2017.12.007
Abstract:
Objective: To investigate the in vitro cytotoxicity of cytotoxic T lymphocytes (CTLs) induced from umbilical cord blood derived dendritic cells (DCs) that sensitized by mammaglobin A (MGBA) (MGBAp-DCs) against breast cancer. Methods: Umbilical cord blood (UCB) samples were obtained from healthy pregnant women during cesarean delivery and used to isolate UCB mononuclear cells to induce DC. MGBAp-DCs were co-cultured with autologous lymphocytes to induce CTL. The DC phenotypes (CD83, CD86, HLA-DR) were determined by using flow cytometry; and IL-10, IL-12 secretion levels was determined by ELISA; the killing effect of CTL against breast cancer cells was detected by CCK-8. Results: Mature DCs were successfully isolated with typical morphology and functionality characteristics. Specific CTL induced by MGBAp- DCs had a significant cytotoxic effect against breast cancer MDA-MB-415 cells (P<0.05). Cytotoxic effect was significantly weakened after adding anti-HLA-I antibody, but not with adding anti-HLA-II antibody. However, the the cytotoxicity of specific CTL on normal breast cell line MCF-10 was not influenced by the presence of anti-HLA-I or anti-HLA-II antibodies or not. Conclusion:MGBAp-DCs can effectively enhance the cytotocxity of CTL against breast cancer cells in vitro.
Objective: To investigate the in vitro cytotoxicity of cytotoxic T lymphocytes (CTLs) induced from umbilical cord blood derived dendritic cells (DCs) that sensitized by mammaglobin A (MGBA) (MGBAp-DCs) against breast cancer. Methods: Umbilical cord blood (UCB) samples were obtained from healthy pregnant women during cesarean delivery and used to isolate UCB mononuclear cells to induce DC. MGBAp-DCs were co-cultured with autologous lymphocytes to induce CTL. The DC phenotypes (CD83, CD86, HLA-DR) were determined by using flow cytometry; and IL-10, IL-12 secretion levels was determined by ELISA; the killing effect of CTL against breast cancer cells was detected by CCK-8. Results: Mature DCs were successfully isolated with typical morphology and functionality characteristics. Specific CTL induced by MGBAp- DCs had a significant cytotoxic effect against breast cancer MDA-MB-415 cells (P<0.05). Cytotoxic effect was significantly weakened after adding anti-HLA-I antibody, but not with adding anti-HLA-II antibody. However, the the cytotoxicity of specific CTL on normal breast cell line MCF-10 was not influenced by the presence of anti-HLA-I or anti-HLA-II antibodies or not. Conclusion:MGBAp-DCs can effectively enhance the cytotocxity of CTL against breast cancer cells in vitro.
2017, 24(12):1386-1390.
DOI: 10.3872/j.issn.1007-385X.2017.12.008
Abstract:
Objective: To investigate the inhibitory effect of Colony stimulating factor- 1 receptor (CSF- 1R) on apoptosis of human nasopharyngeal carcinoma 6-10B cells and its relationship with Bax/Bcl-2 expression. Methods:Lentiviral vector over- expressing CSF-1R LV-CSF1R（16957-1）was constructed and trasfected into 6-10B cells; meanwhile a control group was set; The expression of CSF-1R, Bcl-2 and Bax was detected by Real-time PCR and Western blotting. CCK-8 assay and flow cytometry (FCM) were used to detect the cell proliferation and apoptosis,respectively. Results: The mRNA expression of CSF-1R increased significantly in 6-10B cells of transfection group compared with negative control (NC) group (7.01±0.23 vs 0.09±0.03, P<0.01); The expression of Bax mRNA was significantly down-regulated while the expression of Bcl-2 mRNA was significantly up-regulated in 6-10B cells of transfection group (all P<0.01). The protein expression of CSF-1 in 6-10B cells of transfection group was significantly higher than that of control group, along with the remarkably down- regulated Bax protein expression and slightly up-regulated Bcl-2 protein in transfection group. Compared with NC group, the proliferation vitality of 6-10B cells were extremely increased after transfection (P<0.01); however, the cell apoptosis was significantly reduced ([10.82±0.75]% vs [17.11±0.46]%, P<0.05). Conclusion: Over-expression of CSF-1R could promote the ma-lignant growth and inhibit the apoptosis of nasopharyngeal carcinoma 6-10B cells by regulating the ratio of Bax/Bcl-2 expression.
Objective: To investigate the inhibitory effect of Colony stimulating factor- 1 receptor (CSF- 1R) on apoptosis of human nasopharyngeal carcinoma 6-10B cells and its relationship with Bax/Bcl-2 expression. Methods:Lentiviral vector over- expressing CSF-1R LV-CSF1R（16957-1）was constructed and trasfected into 6-10B cells; meanwhile a control group was set; The expression of CSF-1R, Bcl-2 and Bax was detected by Real-time PCR and Western blotting. CCK-8 assay and flow cytometry (FCM) were used to detect the cell proliferation and apoptosis,respectively. Results: The mRNA expression of CSF-1R increased significantly in 6-10B cells of transfection group compared with negative control (NC) group (7.01±0.23 vs 0.09±0.03, P<0.01); The expression of Bax mRNA was significantly down-regulated while the expression of Bcl-2 mRNA was significantly up-regulated in 6-10B cells of transfection group (all P<0.01). The protein expression of CSF-1 in 6-10B cells of transfection group was significantly higher than that of control group, along with the remarkably down- regulated Bax protein expression and slightly up-regulated Bcl-2 protein in transfection group. Compared with NC group, the proliferation vitality of 6-10B cells were extremely increased after transfection (P<0.01); however, the cell apoptosis was significantly reduced ([10.82±0.75]% vs [17.11±0.46]%, P<0.05). Conclusion: Over-expression of CSF-1R could promote the ma-lignant growth and inhibit the apoptosis of nasopharyngeal carcinoma 6-10B cells by regulating the ratio of Bax/Bcl-2 expression.
2017, 24(12):1391-1396.
DOI: 10.3872/j.issn.1007-385X.2017.12.009
Abstract:
Objective：To investigate the effect of two alternative splicing isoforms of zinc finger protein 148 (ZNF148) gene on the invasion and metastasis of colon cancer cells and their related mechanisms. Methods: RTPCR was used to determine the expressions of two ZNF148 alternative splicing isoforms in the human colorectal SW480 cells. ZNF148 interference vector and ZNF148 over-expression vector were constructed and transfected into the SW480 cells; them were divided into ZNF148FL-siRNA group, ZNF148FL-Over express group, ZNF148ΔN-siRNA group, ZNF148ΔN- Over express group, Control siRNA group, Control Over express group and Normal control group. The mRNA expressions in each group were examined by RT-PCR; the proliferation, invasion and migration in vitro as well as apoptosis of SW480 cells were detected by MTT, Transwell, scratch assay and flow cytometry, respectively.Results: Two splicing isoforms (ZNF148FL of 2 385 bp and ZNF148ΔN of 2 004 bp) were obtained by RTPCR.The expression level of ZNF148FL was significantly decreased while the expression of ZNF148ΔN was increased in the ZNF148FL-siRNA group; the expression of ZNF148FL was significantly increased while the expression of ZNF148ΔN was significantly decreased in the ZNF148FL- Over express group (all P<0.050. The expression of ZNF148ΔN was significantly decreased while the expression of ZNF148FL was increased in ZNF148ΔN-siRNA group;the expression of ZNF148ΔN was significantly increased while the expression of ZNF148FL was decreased in ZNF148ΔN-over express group (all P<0.05). The proliferation of the SW480 cells was increased in ZNF148FL-over express group and the ZNF148ΔN- siRNA group, while the proliferation of the SW480 cells was decreased in the ZNF148FL-siRNA group and the ZNF148ΔN-Over express group. The transmembrane cell number and migration ability of the SW480 cells in the ZNF148FL-siRNA group and the ZNF148ΔN-Over express group were significantly decreased,but the apoptotic rate was significantly increased; However, ZNF148FL-Over Express group and ZNF148ΔNsiRNA group showed the significantly increased transmembrane cell number and migration ability but decreased apoptosis rate (all P<0.05). Conclusion: ZNF148FL could increase proliferation, invasion and metastasis of colorectal cancer cells, while ZNFΔN showed opposite effect; the two splicing isoforms of ZNF148 may exert mutual antagonistic effect to each other on the malignant biological activities.
Objective：To investigate the effect of two alternative splicing isoforms of zinc finger protein 148 (ZNF148) gene on the invasion and metastasis of colon cancer cells and their related mechanisms. Methods: RTPCR was used to determine the expressions of two ZNF148 alternative splicing isoforms in the human colorectal SW480 cells. ZNF148 interference vector and ZNF148 over-expression vector were constructed and transfected into the SW480 cells; them were divided into ZNF148FL-siRNA group, ZNF148FL-Over express group, ZNF148ΔN-siRNA group, ZNF148ΔN- Over express group, Control siRNA group, Control Over express group and Normal control group. The mRNA expressions in each group were examined by RT-PCR; the proliferation, invasion and migration in vitro as well as apoptosis of SW480 cells were detected by MTT, Transwell, scratch assay and flow cytometry, respectively.Results: Two splicing isoforms (ZNF148FL of 2 385 bp and ZNF148ΔN of 2 004 bp) were obtained by RTPCR.The expression level of ZNF148FL was significantly decreased while the expression of ZNF148ΔN was increased in the ZNF148FL-siRNA group; the expression of ZNF148FL was significantly increased while the expression of ZNF148ΔN was significantly decreased in the ZNF148FL- Over express group (all P<0.050. The expression of ZNF148ΔN was significantly decreased while the expression of ZNF148FL was increased in ZNF148ΔN-siRNA group;the expression of ZNF148ΔN was significantly increased while the expression of ZNF148FL was decreased in ZNF148ΔN-over express group (all P<0.05). The proliferation of the SW480 cells was increased in ZNF148FL-over express group and the ZNF148ΔN- siRNA group, while the proliferation of the SW480 cells was decreased in the ZNF148FL-siRNA group and the ZNF148ΔN-Over express group. The transmembrane cell number and migration ability of the SW480 cells in the ZNF148FL-siRNA group and the ZNF148ΔN-Over express group were significantly decreased,but the apoptotic rate was significantly increased; However, ZNF148FL-Over Express group and ZNF148ΔNsiRNA group showed the significantly increased transmembrane cell number and migration ability but decreased apoptosis rate (all P<0.05). Conclusion: ZNF148FL could increase proliferation, invasion and metastasis of colorectal cancer cells, while ZNFΔN showed opposite effect; the two splicing isoforms of ZNF148 may exert mutual antagonistic effect to each other on the malignant biological activities.
2017, 24(12):1397-1402.
DOI: 10.3872/j.issn.1007-385X.2017.12.010
Abstract:
Objective：To investigate the expression patterns of hnRNPA1 mRNA in the digestive system tumors and its clinical significance. Methods：Whole genome mRNA sequencing data of TCGA databases were used to analyze the different expression of hnRNPA1 in tumor and normal tissues and to study its correlation to clinical pathological data; and survival analysis was performed to determine its prognostic value. Results：1. The expressions of hnRNPA1 were significantly higher in digestive system tumor tissues than that in normal tissues（P<0.01），especially in colorectal cancer, hepatocellular carcinoma, pancreatic cancer and cholangiocarcinoma. 2. In hepatocellular carcinoma,hnRNPA1 expression was correlated with tumor differentiation, TNM stage and the depth of invasion (all P<0.05), but not associated with age, sex, risk factors of liver cancer. In pancreatic cancer and cholangiocarcinoma, the expression of hnRNPA1 was not significantly associated with age, sex, TNM stage and infiltration depth etc. 3. The expression of hnRNPA1 was correlated with overall survival in pancreatic cancer patients (P<0.05), which is a risk factor for the prognosis that independent of TNM stage, degree of differentiation; the expression of hnRNPA1 was not significantly associated with overall survival in colorectal cancer, hepatocellular carcinoma and cholangiocarcinoma.Conclusion：The expression of hnRNPA1 mRNA was increased in tumor tissues of different digestive system cancers; its over- expression is an independent risk factor for the prognosis of pancreatic cancer, which might be served as a potential biomarker for predicting prognosis.
Objective：To investigate the expression patterns of hnRNPA1 mRNA in the digestive system tumors and its clinical significance. Methods：Whole genome mRNA sequencing data of TCGA databases were used to analyze the different expression of hnRNPA1 in tumor and normal tissues and to study its correlation to clinical pathological data; and survival analysis was performed to determine its prognostic value. Results：1. The expressions of hnRNPA1 were significantly higher in digestive system tumor tissues than that in normal tissues（P<0.01），especially in colorectal cancer, hepatocellular carcinoma, pancreatic cancer and cholangiocarcinoma. 2. In hepatocellular carcinoma,hnRNPA1 expression was correlated with tumor differentiation, TNM stage and the depth of invasion (all P<0.05), but not associated with age, sex, risk factors of liver cancer. In pancreatic cancer and cholangiocarcinoma, the expression of hnRNPA1 was not significantly associated with age, sex, TNM stage and infiltration depth etc. 3. The expression of hnRNPA1 was correlated with overall survival in pancreatic cancer patients (P<0.05), which is a risk factor for the prognosis that independent of TNM stage, degree of differentiation; the expression of hnRNPA1 was not significantly associated with overall survival in colorectal cancer, hepatocellular carcinoma and cholangiocarcinoma.Conclusion：The expression of hnRNPA1 mRNA was increased in tumor tissues of different digestive system cancers; its over- expression is an independent risk factor for the prognosis of pancreatic cancer, which might be served as a potential biomarker for predicting prognosis.
2017, 24(12):1403-1408.
DOI: 10.3872/j.issn.1007-385X.2017.12.011
Abstract:
Objective: To investigate the expression of microRNA-22 (miR-22) and miR-126 in blood serum of patients with non-small cell lung cancer (NSCLC) and explore their role in the occurrence and metastasis of NSCLC.Methods: A total of 127 NSCLC patients who were diagnosed at the Sixth People's Hospital of Yancheng City during May 2013 till May 2015 were selected in this study, meanwhile another 112 healthy individuals were selected as the control subjects. The qPCR was performed to detect the serum miR-22 and miR-126 levels. Logistic regression analysis was conducted to analyze the independent factors influencing NSCLC metastasis, and receiver operating characteristic (ROC) curve was drawn to analyze the sensitivity and specificity of serum miR-22 and miR-126 levels in predicting NSCLC developments and metastasis. Results: The serum miR-22 level was significantly higher in the case group than that in the control group, while the serum miR-126 level was lower in the case group as compared with that in the control group (all P<0.05). Compared with squamous cell carcinoma patients, serum miR-22 level significantly increased (P<0.05), while serum miR-126 level decreased in patients with adenocarcinoma (P<0.01). Patients at III+IV stage showed increased serum miR-22 level and decreased serum miR-126 level as compared to patients at I + II stage (all P<0.01). In comparison to those without metastasis, patients with metastasis showed elevated serum miR-22 and decreased miR-126 level (all P<0.01). Compared with patients with familial inheritance,those without familial inheritance had increased serum miR-22 level but decreased serum miR-126 level (all P<0.01). The specificity and sensitivity of serum miR-22 and miR-126 levels in predicting NSCLC occurrence were 99.11% and 84.30%, 82.68% and 96.40%, respectively. The specificity and sensitivity of serum miR-22 and miR-126 levels in predicting NSCLC metastasis were 59.74% and 96.00%, 84.00% and 62.30%, respectively. Conclusion:miR-22 was high expressed while miR-126 was low expressed in the serum of NSCLC patients; they may be used as the predicative biomarkers for NSCLC development and metastasis.
Objective: To investigate the expression of microRNA-22 (miR-22) and miR-126 in blood serum of patients with non-small cell lung cancer (NSCLC) and explore their role in the occurrence and metastasis of NSCLC.Methods: A total of 127 NSCLC patients who were diagnosed at the Sixth People's Hospital of Yancheng City during May 2013 till May 2015 were selected in this study, meanwhile another 112 healthy individuals were selected as the control subjects. The qPCR was performed to detect the serum miR-22 and miR-126 levels. Logistic regression analysis was conducted to analyze the independent factors influencing NSCLC metastasis, and receiver operating characteristic (ROC) curve was drawn to analyze the sensitivity and specificity of serum miR-22 and miR-126 levels in predicting NSCLC developments and metastasis. Results: The serum miR-22 level was significantly higher in the case group than that in the control group, while the serum miR-126 level was lower in the case group as compared with that in the control group (all P<0.05). Compared with squamous cell carcinoma patients, serum miR-22 level significantly increased (P<0.05), while serum miR-126 level decreased in patients with adenocarcinoma (P<0.01). Patients at III+IV stage showed increased serum miR-22 level and decreased serum miR-126 level as compared to patients at I + II stage (all P<0.01). In comparison to those without metastasis, patients with metastasis showed elevated serum miR-22 and decreased miR-126 level (all P<0.01). Compared with patients with familial inheritance,those without familial inheritance had increased serum miR-22 level but decreased serum miR-126 level (all P<0.01). The specificity and sensitivity of serum miR-22 and miR-126 levels in predicting NSCLC occurrence were 99.11% and 84.30%, 82.68% and 96.40%, respectively. The specificity and sensitivity of serum miR-22 and miR-126 levels in predicting NSCLC metastasis were 59.74% and 96.00%, 84.00% and 62.30%, respectively. Conclusion:miR-22 was high expressed while miR-126 was low expressed in the serum of NSCLC patients; they may be used as the predicative biomarkers for NSCLC development and metastasis.
2017, 24(12):1409-1413.
DOI: 10.3872/j.issn.1007-385X.2017.12.012
Abstract:
Objective: To investigate the correlation between serum miR-133a level and pathological features of gastric cancer, and to explore the diagnostic and prognostic value of miR-133a. Methods: Ninety-four gastric cancer patients treated in the Department of Gastroenterology Surgery of Affiliated Changzhou N0.2 People’s Hospital of Nanjing Medical University from March 2010 to April 2011 were included in this study; there were 64 male cases and 30 female cases, aged from 32-82 years old with an mean age of (61.72±8.59) years old; Another 30 healthy subjects were enrolled as control. RT-PCR was used to detect the serum miR-133a expression of two groups. SPSS22.0 and Graphpad5.0 were used for statistical analysis. Results: The levels of serum miR-133a in gastric cancer patients were obviously lower than those in healthy controls (1.95±1.51 vs 4.47±0.56, P=0.000). miR-133a expression was significantly associated with the degree of differentiation, depth of invasion, lymph node metastasis and TNM stage (P<0.05), but not obviously associated with age, sex and tumor size (P>0.05). ROC curve analysis showed high sensitivity and specificity of miR-133a in the prediction of gastric cancer (the AUC value of miR-133a was 0.883, 95% CI 0.8257-0.9402) and the prediction of lymph node metastasis (the AUC value of miR-133a was 0.984, 95% CI 0.9633-1.005). The 5-year survival rate of patients with high miR-133a expression was significantly higher than that of low expression group (40.49% vs 19.37%，P<0.05). Conclusion: The expression of miRNA-133a was closely associated with gastric cancer development, its clinicopathological features and patient’s clinical prognosis. It may be used as a potential diagnostic bio-marker and a prognostic predictor for gastric cancer.
Objective: To investigate the correlation between serum miR-133a level and pathological features of gastric cancer, and to explore the diagnostic and prognostic value of miR-133a. Methods: Ninety-four gastric cancer patients treated in the Department of Gastroenterology Surgery of Affiliated Changzhou N0.2 People’s Hospital of Nanjing Medical University from March 2010 to April 2011 were included in this study; there were 64 male cases and 30 female cases, aged from 32-82 years old with an mean age of (61.72±8.59) years old; Another 30 healthy subjects were enrolled as control. RT-PCR was used to detect the serum miR-133a expression of two groups. SPSS22.0 and Graphpad5.0 were used for statistical analysis. Results: The levels of serum miR-133a in gastric cancer patients were obviously lower than those in healthy controls (1.95±1.51 vs 4.47±0.56, P=0.000). miR-133a expression was significantly associated with the degree of differentiation, depth of invasion, lymph node metastasis and TNM stage (P<0.05), but not obviously associated with age, sex and tumor size (P>0.05). ROC curve analysis showed high sensitivity and specificity of miR-133a in the prediction of gastric cancer (the AUC value of miR-133a was 0.883, 95% CI 0.8257-0.9402) and the prediction of lymph node metastasis (the AUC value of miR-133a was 0.984, 95% CI 0.9633-1.005). The 5-year survival rate of patients with high miR-133a expression was significantly higher than that of low expression group (40.49% vs 19.37%，P<0.05). Conclusion: The expression of miRNA-133a was closely associated with gastric cancer development, its clinicopathological features and patient’s clinical prognosis. It may be used as a potential diagnostic bio-marker and a prognostic predictor for gastric cancer.
2017, 24(12):1414-1418.
DOI: 10.3872/j.issn.1007-385X.2017.12.013
Abstract:
Objective: To investigate whether miR- 205- 5p expression in peripheral blood karyocytes from nonsmall cell lung cancer (NSCLC) patients could be used as a reasonable molecule biomarker. Methods: Peripheral blood from 60 patients, who were primarily diagnosed with NSCLC and treated at the Third Affiliated Hospital of Kunming Medical University (Yunnan Cancer Hospital) from June, 2011 to June, 2012, was collected; among them,there were 30 cases with distant metastasis while the other 30 without. In addition, 30 healthy subjects were enrolled as controls. The miR- 205- 5p expressions in collected peripheral blood karyocytes, human bronchial epithelial BEAS-2B cells, Gejiu squamous cell lung carcinoma YTMLC-90 cells, and Xuanwei lung adenocarcinoma XWLC-05 cells were detected by qPCR. We also analyzed the correlation between miR-205- 5p expression in peripheral blood karyocytes and NSCLC serum markers levels, TNM stage, lymph node metastasis, as well as distant metastasis etc. Results: The expressions of miR-205-5p in YTMLC-90 (17.507 2±1.906 3) and XMLC-05 cells (5.725 2±1.012 0) were significantly higher than that in BEAS-2B cells (0.820 8±0.255 3) (P<0.05). The miR-205-5p expres-sion level in peripheral blood karyocytes from healthy controls was significantly higher than that from NSCLC patients (P<0.01). The positive rate of miR-205-5p in peripheral blood karyocytes from NSCLC patients was not correlated with patients’sex, age, lymph node metastasis, pathologic subtypes and tumor serum markers (CEA，CA125,CA242) (all P>0.05), but significantly correlated with distant metastasis, TNM stage and tumor serum marker CA153 (P<0.01). Conclusion: Abnormal up-regulation of miR-205-5p may play an important role in the progression of NSCLC and can be used as a reasonable biomarker for the metastasis in NSCLC patients.
Objective: To investigate whether miR- 205- 5p expression in peripheral blood karyocytes from nonsmall cell lung cancer (NSCLC) patients could be used as a reasonable molecule biomarker. Methods: Peripheral blood from 60 patients, who were primarily diagnosed with NSCLC and treated at the Third Affiliated Hospital of Kunming Medical University (Yunnan Cancer Hospital) from June, 2011 to June, 2012, was collected; among them,there were 30 cases with distant metastasis while the other 30 without. In addition, 30 healthy subjects were enrolled as controls. The miR- 205- 5p expressions in collected peripheral blood karyocytes, human bronchial epithelial BEAS-2B cells, Gejiu squamous cell lung carcinoma YTMLC-90 cells, and Xuanwei lung adenocarcinoma XWLC-05 cells were detected by qPCR. We also analyzed the correlation between miR-205- 5p expression in peripheral blood karyocytes and NSCLC serum markers levels, TNM stage, lymph node metastasis, as well as distant metastasis etc. Results: The expressions of miR-205-5p in YTMLC-90 (17.507 2±1.906 3) and XMLC-05 cells (5.725 2±1.012 0) were significantly higher than that in BEAS-2B cells (0.820 8±0.255 3) (P<0.05). The miR-205-5p expres-sion level in peripheral blood karyocytes from healthy controls was significantly higher than that from NSCLC patients (P<0.01). The positive rate of miR-205-5p in peripheral blood karyocytes from NSCLC patients was not correlated with patients’sex, age, lymph node metastasis, pathologic subtypes and tumor serum markers (CEA，CA125,CA242) (all P>0.05), but significantly correlated with distant metastasis, TNM stage and tumor serum marker CA153 (P<0.01). Conclusion: Abnormal up-regulation of miR-205-5p may play an important role in the progression of NSCLC and can be used as a reasonable biomarker for the metastasis in NSCLC patients.
2017, 24(12):1419-1423.
DOI: 10.3872/j.issn.1007-385X.2017.12.014
Abstract:
Objective: To investigate the relationship between nucleophosmin (NPM, B23, NO38 or NPM1) expression levels in tumor biopsy tissues as well as the proliferation index and the therapeutic efficacy in nasopharyngeal carcinoma (NPC) patients. Methods: One hundred and fifty- seven nasopharyngeal carcinoma patients, who were treated at Department of Pathology, Sun Yat-sen University cancer center during the period from January 1, 2001 till December 31, 2003 with preserved pre-treatment paraffin-embedded biopsy blocks, were randomly selected. The expressions of Ki-67 and NPM1 in the 157 sample tissues were determined by immunohistochemistry, respectively.The clinical data of patients were reviewed in detail, and the SPSS-16 soft ware was adopted for statistical analysis.Results: The levels of NPM1 were positively correlated with Ki-67 expression (r=0.445, P<0.01), but negatively correlated with regional lymph node status (N0, N1, N2, and N3; r=-0.235, P<0.01); however, it’s not correlated with primary tumor staging (T1, T2, T3 and T4; r=0.006, P>0.05) or clinical staging (r=-0.74, P>0.05). The expression of NPM1 was negatively correlated with the palpable rate of lymph node(s) (r=-0.196, P<0.05) in lymph node metastasis group, and negatively correlated with the primary lesion status after treatment (r=-0.349, P<0.05)in lymph node non-metastasis group. NPM1 expression was not significantly correlated with of the prognosis of patients. Conclusion: NPM1 expression in nasopharyngeal carcinoma tissues was positively correlated with Ki67 expression, proving NPM1 could promote the cell proliferation and might be used as an indicator for predicting the efficacy of individualized therapy for nasopharyngeal carcinoma patients.
Objective: To investigate the relationship between nucleophosmin (NPM, B23, NO38 or NPM1) expression levels in tumor biopsy tissues as well as the proliferation index and the therapeutic efficacy in nasopharyngeal carcinoma (NPC) patients. Methods: One hundred and fifty- seven nasopharyngeal carcinoma patients, who were treated at Department of Pathology, Sun Yat-sen University cancer center during the period from January 1, 2001 till December 31, 2003 with preserved pre-treatment paraffin-embedded biopsy blocks, were randomly selected. The expressions of Ki-67 and NPM1 in the 157 sample tissues were determined by immunohistochemistry, respectively.The clinical data of patients were reviewed in detail, and the SPSS-16 soft ware was adopted for statistical analysis.Results: The levels of NPM1 were positively correlated with Ki-67 expression (r=0.445, P<0.01), but negatively correlated with regional lymph node status (N0, N1, N2, and N3; r=-0.235, P<0.01); however, it’s not correlated with primary tumor staging (T1, T2, T3 and T4; r=0.006, P>0.05) or clinical staging (r=-0.74, P>0.05). The expression of NPM1 was negatively correlated with the palpable rate of lymph node(s) (r=-0.196, P<0.05) in lymph node metastasis group, and negatively correlated with the primary lesion status after treatment (r=-0.349, P<0.05)in lymph node non-metastasis group. NPM1 expression was not significantly correlated with of the prognosis of patients. Conclusion: NPM1 expression in nasopharyngeal carcinoma tissues was positively correlated with Ki67 expression, proving NPM1 could promote the cell proliferation and might be used as an indicator for predicting the efficacy of individualized therapy for nasopharyngeal carcinoma patients.
2017, 24(12):1424-1430.
DOI: 10.3872/j.issn.1007-385X.2017.12.015
Abstract:
磷脂酰肌醇3-蛋白激酶B-哺乳动物雷帕霉素靶蛋白（PI3K-AKT-mTOR）信号通路的上游通路主要包括表皮生长因子受体(EGFR)、人类表皮生长因子受体2(HER2)、肝细胞生长因子受体(MET)和成纤维细胞生长因子受体(FGFR)等各类受体的酪氨酸激酶，其受体的突变、扩增常会导致PI3K-AKT-mTOR信号通路的失调，引起肿瘤发生发展。该通路异常激活常伴随着下游关键分子的改变包括PIK3CA、AKT1、PTEN 的基因突变，PIK3CA、AKT1、AKT2 的基因扩增，肿瘤抑制基因PTEN 的缺失等。PI3K-AKT-mTOR信号通路抑制剂主要包括PI3K抑制剂、AKT抑制剂、mTOR抑制剂和双重抑制剂等。PI3K-AKT-mTOR抑制剂对肿瘤免疫微环境、免疫细胞均有显著影响。PI3K-AKT-mTOR抑制剂单药治疗肿瘤具有一定局限性，如联合DC疫苗、免疫检查点抑制剂和CAR-T细胞等免疫疗法可增强抗肿瘤疗效。
磷脂酰肌醇3-蛋白激酶B-哺乳动物雷帕霉素靶蛋白（PI3K-AKT-mTOR）信号通路的上游通路主要包括表皮生长因子受体(EGFR)、人类表皮生长因子受体2(HER2)、肝细胞生长因子受体(MET)和成纤维细胞生长因子受体(FGFR)等各类受体的酪氨酸激酶，其受体的突变、扩增常会导致PI3K-AKT-mTOR信号通路的失调，引起肿瘤发生发展。该通路异常激活常伴随着下游关键分子的改变包括PIK3CA、AKT1、PTEN 的基因突变，PIK3CA、AKT1、AKT2 的基因扩增，肿瘤抑制基因PTEN 的缺失等。PI3K-AKT-mTOR信号通路抑制剂主要包括PI3K抑制剂、AKT抑制剂、mTOR抑制剂和双重抑制剂等。PI3K-AKT-mTOR抑制剂对肿瘤免疫微环境、免疫细胞均有显著影响。PI3K-AKT-mTOR抑制剂单药治疗肿瘤具有一定局限性，如联合DC疫苗、免疫检查点抑制剂和CAR-T细胞等免疫疗法可增强抗肿瘤疗效。
2017, 24(12):1431-1435.
DOI: 10.3872/j.issn.1007-385X.2017.12.016
Abstract:
癌相关成纤维细胞（cancer associated fibroblasts, CAF）在肿瘤微环境中起着重要作用，其不仅能影响正常细胞的恶性转化，还促进肿瘤细胞的增殖、侵袭及远处转移。CAF通过物理吸引作用促进癌细胞转移、促进肿瘤血管生成，影响肿瘤细胞的代谢，从而促进恶性侵袭。外泌体在微环境中能够从供体细胞向受体细胞传递分子和遗传物质，是分子在细胞间转移的载体，在CAF与恶性肿瘤细胞沟通中发挥重要作用。CAF分泌外泌体促进肿瘤的发展，肿瘤细胞源性外泌体能激活CAF。外泌体具有作为靶向药物递送载体的潜在作用，外泌体作为生物特异性标志物也可用于癌症的早期检测、诊断和预后，具有较好的临床应用前景。
癌相关成纤维细胞（cancer associated fibroblasts, CAF）在肿瘤微环境中起着重要作用，其不仅能影响正常细胞的恶性转化，还促进肿瘤细胞的增殖、侵袭及远处转移。CAF通过物理吸引作用促进癌细胞转移、促进肿瘤血管生成，影响肿瘤细胞的代谢，从而促进恶性侵袭。外泌体在微环境中能够从供体细胞向受体细胞传递分子和遗传物质，是分子在细胞间转移的载体，在CAF与恶性肿瘤细胞沟通中发挥重要作用。CAF分泌外泌体促进肿瘤的发展，肿瘤细胞源性外泌体能激活CAF。外泌体具有作为靶向药物递送载体的潜在作用，外泌体作为生物特异性标志物也可用于癌症的早期检测、诊断和预后，具有较好的临床应用前景。
2017, 24(12):1436-1442.
DOI: 10.3872/j.issn.1007-385X.2017.12.017
Abstract:
转录信号转导子与激活子3（STAT3）持续活化及异常高表达，与肿瘤的发生发展及不良预后密切相关，已成为新型抗瘤药物的潜在靶标。目前，靶向STAT3 信号转导通路的肿瘤治疗策略主要包括小分子干扰RNA、特异性寡核苷酸、正向调控促癌基因敲除等核酸类抑制干预，JAK-2 抑制剂、酪氨酸及其激酶抑制剂、细胞因子及受体拮抗剂等STAT3 上游信号阻断及负调控蛋白表达促进，STAT3 负显性蛋白、二聚体拮抗剂等STAT3/pSTAT3 特异性抑制。另外，天然药物有效成分及其他具有靶向STAT3信号的药剂，亦可通过下调STAT3 蛋白水平、抑制蛋白磷酸化、靶向STAT3 上游信号转导及下游靶分子阻抑肿瘤细胞生长增殖和侵袭转移。
转录信号转导子与激活子3（STAT3）持续活化及异常高表达，与肿瘤的发生发展及不良预后密切相关，已成为新型抗瘤药物的潜在靶标。目前，靶向STAT3 信号转导通路的肿瘤治疗策略主要包括小分子干扰RNA、特异性寡核苷酸、正向调控促癌基因敲除等核酸类抑制干预，JAK-2 抑制剂、酪氨酸及其激酶抑制剂、细胞因子及受体拮抗剂等STAT3 上游信号阻断及负调控蛋白表达促进，STAT3 负显性蛋白、二聚体拮抗剂等STAT3/pSTAT3 特异性抑制。另外，天然药物有效成分及其他具有靶向STAT3信号的药剂，亦可通过下调STAT3 蛋白水平、抑制蛋白磷酸化、靶向STAT3 上游信号转导及下游靶分子阻抑肿瘤细胞生长增殖和侵袭转移。
2017, 24(12):1443-1448.
DOI: 10.3872/j.issn.1007-385X.2017.12.018
Abstract:
通过免疫检查点分子和配体结合而激活的免疫检查点通路是活化免疫细胞的抑制性第二信号，维持机体自身免疫耐受，抑制免疫过度激活从而保护自身组织。其中主要包括CTLA-4/CD80、CD86，PD-1/PD-L1，A2aR/细胞外腺苷和Tim-3/半乳凝素-9 等。乳腺癌细胞通过高表达免疫检查点分子配体介导免疫逃逸，通过PI3K通路、JAK/STAT通路和NF-κB通路参与免疫作用机制。目前，针对免疫检查点的乳腺癌治疗策略包括直接阻断T细胞表面的免疫检查点分子，如单克隆抗体单一治疗、单克隆抗体间联合治疗、单克隆抗体联合化疗药物治疗、单克隆抗体联合物理疗法、基因靶向治疗等，以及间接阻断肿瘤细胞及其微环境的免疫检查点分子配体，其研究热点主要集中在PD-L1上，具体方法主要为单克隆抗体以及基因靶向治疗。
通过免疫检查点分子和配体结合而激活的免疫检查点通路是活化免疫细胞的抑制性第二信号，维持机体自身免疫耐受，抑制免疫过度激活从而保护自身组织。其中主要包括CTLA-4/CD80、CD86，PD-1/PD-L1，A2aR/细胞外腺苷和Tim-3/半乳凝素-9 等。乳腺癌细胞通过高表达免疫检查点分子配体介导免疫逃逸，通过PI3K通路、JAK/STAT通路和NF-κB通路参与免疫作用机制。目前，针对免疫检查点的乳腺癌治疗策略包括直接阻断T细胞表面的免疫检查点分子，如单克隆抗体单一治疗、单克隆抗体间联合治疗、单克隆抗体联合化疗药物治疗、单克隆抗体联合物理疗法、基因靶向治疗等，以及间接阻断肿瘤细胞及其微环境的免疫检查点分子配体，其研究热点主要集中在PD-L1上，具体方法主要为单克隆抗体以及基因靶向治疗。
联系方式
- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
- 邮编：200433
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.