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Volume 25,Issue 3,2018 Table of Contents
2018, 25(3):213-220.
DOI: 10.3872/j.issn.1007-385X.2018.03.001
Abstract:
The research on immune checkpoint has made breakthrough progress in recent years. The PD-1/PD-L1 signaling pathway is closely related to the immune escape mechanism, and their inhibitors have also made great success in lung cancer. From Checkmate- 017, Checkmate-057 to KEYNOTE-010 and OAK studies, PD-1/PD-L1 inhibitors have gradually established their position as standard treatment for the advanced NSCLC patients after chemotherapy failed. PD-1/PD-L1 inhibitors can be combined with other cancer treat- ment methods, including radiotherapy, chemotherapy, targeted therapy and other immunotherapies, whichhave synergistic effects in the treatment of lung cancer, thus improving the efficacy. Immune checkpoint inhibitors bring changes in the treatment patterns of lung can- cer, but also to the traditional efficacy evaluation model,as wellas reaction to the treatment-related adverse. In addition, the development of immune check point inhibitors has effectively promoted the progress of precision medicine.
The research on immune checkpoint has made breakthrough progress in recent years. The PD-1/PD-L1 signaling pathway is closely related to the immune escape mechanism, and their inhibitors have also made great success in lung cancer. From Checkmate- 017, Checkmate-057 to KEYNOTE-010 and OAK studies, PD-1/PD-L1 inhibitors have gradually established their position as standard treatment for the advanced NSCLC patients after chemotherapy failed. PD-1/PD-L1 inhibitors can be combined with other cancer treat- ment methods, including radiotherapy, chemotherapy, targeted therapy and other immunotherapies, whichhave synergistic effects in the treatment of lung cancer, thus improving the efficacy. Immune checkpoint inhibitors bring changes in the treatment patterns of lung can- cer, but also to the traditional efficacy evaluation model,as wellas reaction to the treatment-related adverse. In addition, the development of immune check point inhibitors has effectively promoted the progress of precision medicine.
2018, 25(3):221-228.
DOI: 10.3872/j.issn.1007-385X.2018.03.002
Abstract:
Objective: To investigate the effect of sphingosine kinase 1 (SphK1) knockdown on the proliferation and migration of co- lon cancer RKO cells induced by mesenchymal stem cells (MSCs). Methods: RKO cells were treated with MSCs conditioned medium (MSC-CM) or control medium (Control-CM), respectively. Cell proliferation was detected by CCK-8 assay. Cell migration ability was tested by Transwell chamber assay. The proteins expression of Ki-67, MMP-2/9, CD44 and CD133 was detected by Western blotting. Then, the expression of SphK1 in RKO cells was suppressed by targeted gene lentivirus shRNA vector transfection. The effects of SphK1 knockdown on the proliferation, migration and protein expressions of Ki-67, MMP-2/9, CD44 and CD133 of RKO cells induced by MSC-CM were observed. Results: The RKO cells proliferation was promoted by MSC-CM in a time-dependent manner; moreover (P<0.05), the migration ability of cells was significantly enhanced after being treated with MSC-CM(P<0.01). In addition, MSC-CM significantly increased the protein expressions of Ki-67, MMP-2/9, CD44 and CD133(all P<0.05 or P<0.01). Lentiviral ShRNA vector transfection could significantly inhibit the expression of SphK1. Down-regulation of SphK1 significantly inhibited the proliferation, mi- gration and protein expressions of Ki-67, MMP-2/9, CD44 a D133 of RKO cells induced by MSC-CM(all P<0.05 or P<0.01). Con- clusion: MSC-CM promotes the proliferation and migration of colon cancer RKO cells. Down-regulation of SphK1 reverses the cell proliferation and migration induced by MSC-CM via inhibiting the expression of MMP-2/9, CD44 and CD133.
Objective: To investigate the effect of sphingosine kinase 1 (SphK1) knockdown on the proliferation and migration of co- lon cancer RKO cells induced by mesenchymal stem cells (MSCs). Methods: RKO cells were treated with MSCs conditioned medium (MSC-CM) or control medium (Control-CM), respectively. Cell proliferation was detected by CCK-8 assay. Cell migration ability was tested by Transwell chamber assay. The proteins expression of Ki-67, MMP-2/9, CD44 and CD133 was detected by Western blotting. Then, the expression of SphK1 in RKO cells was suppressed by targeted gene lentivirus shRNA vector transfection. The effects of SphK1 knockdown on the proliferation, migration and protein expressions of Ki-67, MMP-2/9, CD44 and CD133 of RKO cells induced by MSC-CM were observed. Results: The RKO cells proliferation was promoted by MSC-CM in a time-dependent manner; moreover (P<0.05), the migration ability of cells was significantly enhanced after being treated with MSC-CM(P<0.01). In addition, MSC-CM significantly increased the protein expressions of Ki-67, MMP-2/9, CD44 and CD133(all P<0.05 or P<0.01). Lentiviral ShRNA vector transfection could significantly inhibit the expression of SphK1. Down-regulation of SphK1 significantly inhibited the proliferation, mi- gration and protein expressions of Ki-67, MMP-2/9, CD44 a D133 of RKO cells induced by MSC-CM(all P<0.05 or P<0.01). Con- clusion: MSC-CM promotes the proliferation and migration of colon cancer RKO cells. Down-regulation of SphK1 reverses the cell proliferation and migration induced by MSC-CM via inhibiting the expression of MMP-2/9, CD44 and CD133.
3 Knock-down of cytokine induced apoptosis inhibitor 1 gene sensitized leukemia K562
cells to imatinib
2018, 25(3):229-235.
DOI: 10.3872/j.issn.1007-385X.2018.03.003
Abstract:
Objective: To investigate the biological effects and the related mechanisms of cytokine induced apoptosis inhibitor 1 (CIAPIN1) on the sensitivity of K562 chronic myeloid leukemia cells to imatinib. Methods: Specific short hairpin RNA (shRNA) inter- ference vectors targeting CIAPIN1 (CIAPIN1-shRNA) were constructed. Interference efficiency of interference group (K562 cells trans- fected with CIAPIN1-shRNA) and control group (K562 cells transfected with scramble-shRNA) was evaluated using Real-time PCR, Western blotting and immunofluorescence staining. The interference group and control group were treated by 2 μmol/L imatinib. Cell viability was detected using MTT assay. Colony formation ability was detected using cell colony forming experiment. Cell cycle and apoptosis was detected using Flow cytometry and Western blotting. Results: CIAPIN1 expression was decreased effectively by specific shRNA targeting CIAPIN1. The CIAPIN1 mRNA content in CIAPIN1-shRNA group accounted (29.74±4.03)% of scramble-shRNA group, while the CIAPIN1 protein content in CIAPIN1-shRNA group accounted (21.57±2.18)% of scramble-shRNA group. CIAPIN1 knock-down significantly enhanced the inhibitory activity of imatinib on proliferation and colony forming ability of K562 cells. The col-ony number and radius of the CIAPIN1-shRNA+imatinib group was (15.60±1.03) and (2.63±0.55) μm, which were all less than those of the scramble-shRNA+imatinib group. The knock down also increased the activity of imatinib to block the cell cycle at G1 phase and to promot apoptosis of cells. The cell ratio at G1 phase of the CIAPIN1-shRNA+imatinib group was obviously increased while the ratio at S phase was obviously decreased compared with those of scramble-shRNA+imatinib group. Hoechst33258 staining and flow cytome- try showed that the proportion of apoptotic K562 cells in the CIAPIN1-shRNA+imatinib group increased. The results of Western blot- ting showed that CIAPIN1 knock-down not only up-regulated the expressions of apoptosis related proteins (p21, Bid and Bim), but also repressed expressions of cell cycle related proteins (Cyclin D1, Bcl-xl, Bcl-2 and Mcl-1), which had synergistic effects with imatinib. Conclusion: CIAPIN1 knock-down significantly sensitized K562 cells to imatinib treatment, and the mechanism might be related with cell cycle arrest and expression of apoptosis-associated proteins.
Objective: To investigate the biological effects and the related mechanisms of cytokine induced apoptosis inhibitor 1 (CIAPIN1) on the sensitivity of K562 chronic myeloid leukemia cells to imatinib. Methods: Specific short hairpin RNA (shRNA) inter- ference vectors targeting CIAPIN1 (CIAPIN1-shRNA) were constructed. Interference efficiency of interference group (K562 cells trans- fected with CIAPIN1-shRNA) and control group (K562 cells transfected with scramble-shRNA) was evaluated using Real-time PCR, Western blotting and immunofluorescence staining. The interference group and control group were treated by 2 μmol/L imatinib. Cell viability was detected using MTT assay. Colony formation ability was detected using cell colony forming experiment. Cell cycle and apoptosis was detected using Flow cytometry and Western blotting. Results: CIAPIN1 expression was decreased effectively by specific shRNA targeting CIAPIN1. The CIAPIN1 mRNA content in CIAPIN1-shRNA group accounted (29.74±4.03)% of scramble-shRNA group, while the CIAPIN1 protein content in CIAPIN1-shRNA group accounted (21.57±2.18)% of scramble-shRNA group. CIAPIN1 knock-down significantly enhanced the inhibitory activity of imatinib on proliferation and colony forming ability of K562 cells. The col-ony number and radius of the CIAPIN1-shRNA+imatinib group was (15.60±1.03) and (2.63±0.55) μm, which were all less than those of the scramble-shRNA+imatinib group. The knock down also increased the activity of imatinib to block the cell cycle at G1 phase and to promot apoptosis of cells. The cell ratio at G1 phase of the CIAPIN1-shRNA+imatinib group was obviously increased while the ratio at S phase was obviously decreased compared with those of scramble-shRNA+imatinib group. Hoechst33258 staining and flow cytome- try showed that the proportion of apoptotic K562 cells in the CIAPIN1-shRNA+imatinib group increased. The results of Western blot- ting showed that CIAPIN1 knock-down not only up-regulated the expressions of apoptosis related proteins (p21, Bid and Bim), but also repressed expressions of cell cycle related proteins (Cyclin D1, Bcl-xl, Bcl-2 and Mcl-1), which had synergistic effects with imatinib. Conclusion: CIAPIN1 knock-down significantly sensitized K562 cells to imatinib treatment, and the mechanism might be related with cell cycle arrest and expression of apoptosis-associated proteins.
2018, 25(3):236-239.
DOI: 10.3872/j.issn.1007-385X.2018.03.004
Abstract:
Objective: To investigate he effect of tetracycline- (Tet-on) mediated livin RNA interference on growth of lung carcinoma xenegrafts, and find a better regulatory way to interfere the development on lung cancer. Methods: livin shRNA lentiviral vectors were constructed; and the lung cancerA549 cells were subcutaneously injected into right upper back of nude mice to establish xenegraft mod- el. The livin shRNA lentiviral vectors were injected into xenografts to interfere the expression of livin, then tetracycline was injected in- traperitoneally for the induction. The suppressive effect of Tet-on mediated livin RNA interference efficiency was investigated and lung cancer xenograft development was observed. Results: After the induction with Tet-on, livin gene expression was significantly inhibited by livin shRNA compared with the control group and Tet-on-NC group; the xenograft volume in Tet-on- livin shRNA group was signifi- cantly smaller than that in control group and Tet-on-NC group ([5.31±0.86]g vs [8.22±0.63]g and [7.17±0.54] g, P<0.05). Moreover, lit- tle body toxicity was observed and no nude mice died in this study. Conclusion: The Tet-on mediated livin shRNA could suppress the growth of lung cancer development with good targeting and controllable characteristics, which might provide a potent tool for treating lung cancer with livin protein as target.
Objective: To investigate he effect of tetracycline- (Tet-on) mediated livin RNA interference on growth of lung carcinoma xenegrafts, and find a better regulatory way to interfere the development on lung cancer. Methods: livin shRNA lentiviral vectors were constructed; and the lung cancerA549 cells were subcutaneously injected into right upper back of nude mice to establish xenegraft mod- el. The livin shRNA lentiviral vectors were injected into xenografts to interfere the expression of livin, then tetracycline was injected in- traperitoneally for the induction. The suppressive effect of Tet-on mediated livin RNA interference efficiency was investigated and lung cancer xenograft development was observed. Results: After the induction with Tet-on, livin gene expression was significantly inhibited by livin shRNA compared with the control group and Tet-on-NC group; the xenograft volume in Tet-on- livin shRNA group was signifi- cantly smaller than that in control group and Tet-on-NC group ([5.31±0.86]g vs [8.22±0.63]g and [7.17±0.54] g, P<0.05). Moreover, lit- tle body toxicity was observed and no nude mice died in this study. Conclusion: The Tet-on mediated livin shRNA could suppress the growth of lung cancer development with good targeting and controllable characteristics, which might provide a potent tool for treating lung cancer with livin protein as target.
2018, 25(3):240-245.
DOI: 10.3872/j.issn.1007-385X.2018.03.005
Abstract:
Objective: To establish paclitaxel(PTX)-resistant human triple negative breast cancer cell line and to examine the expres- sion profile of FA-related genes and FANCF, the correlation between the expression of FA-related genes, FANCF and PTX-resistance in breast cancer were further analyzed. Methods: PTX-resistant MDA-MB-231 cell line was established by means of long-term PTX-ex- posed culture. The sensitivity of the cells to paclitaxel was determined by the CCK8 assay. The cell cycle distribution was examined by flow cytometry after exposure to the paclitaxel. The expression of FA-related gene mRNA and FANCF protein were examined by using real time quantitative PCR and Western blotting. The expression of FANCF in the cells was reduced by RNAi interference technology and the effect of the RNAi was verified. Results: MDA-MB-231/PTX cell showed a 9.9-fold resistance to paclitaxel, indicating that the cell had acquired resistance to PTX. PTX treatment significantly induced G0/G1 arrest and the number of cells in phase S markedly de- creased after exposure to PTX. The mRNA and protein expression of FANCF was significantly higher in PTX-resistant cell than that in PTX-sensitve parental cell，Knockdown of FANCF induced apoptosis in MDA-MB-231/PTX cell as well as in parental cell. FANCF knockdown increased the sensitivity of paclitaxel to both MDA-MB-231 and MDA-MB-231/PTX cells (P<0.05 or P<0.01). Conclu- sion: FANCF played an important role in PTX resistance of the breast cancer cells and FANCF might be a target for therapy aimed at re- versing chemoresistance.
Objective: To establish paclitaxel(PTX)-resistant human triple negative breast cancer cell line and to examine the expres- sion profile of FA-related genes and FANCF, the correlation between the expression of FA-related genes, FANCF and PTX-resistance in breast cancer were further analyzed. Methods: PTX-resistant MDA-MB-231 cell line was established by means of long-term PTX-ex- posed culture. The sensitivity of the cells to paclitaxel was determined by the CCK8 assay. The cell cycle distribution was examined by flow cytometry after exposure to the paclitaxel. The expression of FA-related gene mRNA and FANCF protein were examined by using real time quantitative PCR and Western blotting. The expression of FANCF in the cells was reduced by RNAi interference technology and the effect of the RNAi was verified. Results: MDA-MB-231/PTX cell showed a 9.9-fold resistance to paclitaxel, indicating that the cell had acquired resistance to PTX. PTX treatment significantly induced G0/G1 arrest and the number of cells in phase S markedly de- creased after exposure to PTX. The mRNA and protein expression of FANCF was significantly higher in PTX-resistant cell than that in PTX-sensitve parental cell，Knockdown of FANCF induced apoptosis in MDA-MB-231/PTX cell as well as in parental cell. FANCF knockdown increased the sensitivity of paclitaxel to both MDA-MB-231 and MDA-MB-231/PTX cells (P<0.05 or P<0.01). Conclu- sion: FANCF played an important role in PTX resistance of the breast cancer cells and FANCF might be a target for therapy aimed at re- versing chemoresistance.
2018, 25(3):246-251.
DOI: 10.3872/j.issn.1007-385X.2018.03.006
Abstract:
Objective: To investigate the miRNAs that can intervene Mcl-1 expression in HBV-related liver cancers and to study their synergistic anti-cancer effect with sorafenib. Methods: The expressions of miR-29, miR-101 and miR-193b in HepG2.2.15 (HBV posi- tive) and HepG2.vc (HBV negative) cells were detected by qPCR. miRNA mimics of low expressed genes in HepG2.2.15 cells were synthesized and transfected into HepG2.2.15 and HepG2.vc cells, respectively. qPCR was used to detect target miRNA expression. Western blotting was used to detect the expression of mcl-1 protein in cells before and after transfection. At the same time, (1×10 -9 )~(1× 10 -3 ) mol/L of sorafenib was add to both transfected and non-transfected HepG2.2.15 and HepG2.vc cells; 72 h later, the IC 50 and cell apoptosis was evaluated. Results: The expression of miR-193b in HepG2.2.15 cells was significantly lower than that in HepG2.2.15 cells (P <0.05). The expression of miR-193b in HepG2.2.15 cells and HepG2.2.15 cells was significantly higher after miR-193b mimics transfection (P <0.05). Compared with HepG2.vc cells, the expression of Mcl-1 protein in HepG2.2.15 cells was significantly increased (P <0.05). The expression of Mcl-1 protein in HepG2.2.15 and HepG2.vc cells was significantly decreased after miR-193b mimics transfection (P<0.05). After miR-193b mimics transfection, sorafenib could significantly increase apoptosis rate of both HepG2.2.15 and HepG2.vc cells. Conclusion: The low susceptibility of HBV-related liver cancer to sorafenib may be related with the low expres-sion of miR-193b in cancer cells. Mcl-1 might be used as a target of miR-193b, and miR-193b mimics have a significant synergistic ef- fect with sorafenib.
Objective: To investigate the miRNAs that can intervene Mcl-1 expression in HBV-related liver cancers and to study their synergistic anti-cancer effect with sorafenib. Methods: The expressions of miR-29, miR-101 and miR-193b in HepG2.2.15 (HBV posi- tive) and HepG2.vc (HBV negative) cells were detected by qPCR. miRNA mimics of low expressed genes in HepG2.2.15 cells were synthesized and transfected into HepG2.2.15 and HepG2.vc cells, respectively. qPCR was used to detect target miRNA expression. Western blotting was used to detect the expression of mcl-1 protein in cells before and after transfection. At the same time, (1×10 -9 )~(1× 10 -3 ) mol/L of sorafenib was add to both transfected and non-transfected HepG2.2.15 and HepG2.vc cells; 72 h later, the IC 50 and cell apoptosis was evaluated. Results: The expression of miR-193b in HepG2.2.15 cells was significantly lower than that in HepG2.2.15 cells (P <0.05). The expression of miR-193b in HepG2.2.15 cells and HepG2.2.15 cells was significantly higher after miR-193b mimics transfection (P <0.05). Compared with HepG2.vc cells, the expression of Mcl-1 protein in HepG2.2.15 cells was significantly increased (P <0.05). The expression of Mcl-1 protein in HepG2.2.15 and HepG2.vc cells was significantly decreased after miR-193b mimics transfection (P<0.05). After miR-193b mimics transfection, sorafenib could significantly increase apoptosis rate of both HepG2.2.15 and HepG2.vc cells. Conclusion: The low susceptibility of HBV-related liver cancer to sorafenib may be related with the low expres-sion of miR-193b in cancer cells. Mcl-1 might be used as a target of miR-193b, and miR-193b mimics have a significant synergistic ef- fect with sorafenib.
2018, 25(3):252-257.
DOI: 10.3872/j.issn.1007-385X.2018.03.007
Abstract:
Objective: To study the effects of ursolic acid cooperated with gemcitabine on proliferation and apoptosis of pancreatic can- cer PANC-1 cells. Methods: Human pancreatic cancer cell line PANC-1 was cultured in vitro with ursolic acid and gemcitabine respec- tively; and MTT assay was used to determine the IC 50 of ursolic acid and gemcitabine, thus obtaining the best drug concentration. Urso- lic acid (2 μ mol/L) and gemcitabine (0.2 μ mol/L) alone or in combination was used to treat PANC-1 cells; trypan blue assay was used to test cell viability, and PI staining was used to examine the cell apoptosis; wound healing was used to detect the proliferation and mi- gration of PANC-1 cells. The protein expressions of P-JNK, Bcl-2, IL-6, P-Stat 3, NF-κB and Cox-2 in cells of each treatment group were detected using Western blotting. Results: Both ursolic acid and gemcitabine could significantly inhibit the proliferation of PANC-1 cells, and the IC 50 is 13.67 and 2.78 μ mol/L, respectively; and the final concentrations were determined at 2 and 0.2 μ mol/L for ursolic acid and gemcitabine, respectively. Compared with single drug treatment, the combined treatment exerted a more prominent cell proliferation inhibition effect ([46.47±5.07]% vs [78.38±8.65]%, [76.12±3.23]%, all P<0.05), apoptosis-induction effect ([39.78± 7.01]% vs [20.35±8.51]%, [20.35±8.51]%, all P<0.01) and migration inhibition effect (P<0.01) on PANC-1 cells. Western blotting showed that the combined treatment strongly inhibited Bcl-2 and IL-6 expression, accelerated P-JNK protein expression compared with single drug treatment. Conclusion: The synergistic effect of ursolic acid and gemcitabine enhanced the inhibition on proliferation, mi- gration, and promoted cell apoptosis of human pancreatic cancer cell line PANC-1, the mechanism may be associated with inhibition of Bcl-2, Il-6, P-stat 3 proteins and promotion of P-JNK protein.
Objective: To study the effects of ursolic acid cooperated with gemcitabine on proliferation and apoptosis of pancreatic can- cer PANC-1 cells. Methods: Human pancreatic cancer cell line PANC-1 was cultured in vitro with ursolic acid and gemcitabine respec- tively; and MTT assay was used to determine the IC 50 of ursolic acid and gemcitabine, thus obtaining the best drug concentration. Urso- lic acid (2 μ mol/L) and gemcitabine (0.2 μ mol/L) alone or in combination was used to treat PANC-1 cells; trypan blue assay was used to test cell viability, and PI staining was used to examine the cell apoptosis; wound healing was used to detect the proliferation and mi- gration of PANC-1 cells. The protein expressions of P-JNK, Bcl-2, IL-6, P-Stat 3, NF-κB and Cox-2 in cells of each treatment group were detected using Western blotting. Results: Both ursolic acid and gemcitabine could significantly inhibit the proliferation of PANC-1 cells, and the IC 50 is 13.67 and 2.78 μ mol/L, respectively; and the final concentrations were determined at 2 and 0.2 μ mol/L for ursolic acid and gemcitabine, respectively. Compared with single drug treatment, the combined treatment exerted a more prominent cell proliferation inhibition effect ([46.47±5.07]% vs [78.38±8.65]%, [76.12±3.23]%, all P<0.05), apoptosis-induction effect ([39.78± 7.01]% vs [20.35±8.51]%, [20.35±8.51]%, all P<0.01) and migration inhibition effect (P<0.01) on PANC-1 cells. Western blotting showed that the combined treatment strongly inhibited Bcl-2 and IL-6 expression, accelerated P-JNK protein expression compared with single drug treatment. Conclusion: The synergistic effect of ursolic acid and gemcitabine enhanced the inhibition on proliferation, mi- gration, and promoted cell apoptosis of human pancreatic cancer cell line PANC-1, the mechanism may be associated with inhibition of Bcl-2, Il-6, P-stat 3 proteins and promotion of P-JNK protein.
2018, 25(3):258-262.
DOI: 10.3872/j.issn.1007-385X.2018.03.008
Abstract:
Objective： ：The co-immunoprecipitation and mass spectrometric analysis was carried out to obtain the S-phase kinase-asso- ciated protein 2 (SKP2)-binding proteins in HeLa cells, and the biological functions of these binding proteins were forecast. Methods: The co-immunoprecipitation system was established by co-immunoprecipitation and Western blotting assay; the specific protein gel of SKP2-binding proteins was obtained by SDS-PAGE and silver staining assay; the potential SKP2-binding proteins was identified by mass spectrometric analysis; and the GO (Gene ontology) analysis and KEGG analysis was carried out by bioinformatics technique. Results: The expression level of SKP2 protein in HeLa cells was high enough for co-immunoprecipitation assay; the co-immunoprecipi- tation system was established successfully, and SKP2-binding proteins was obtained; a total of 563 proteins were identified by mass spectrometric analysis, and 270 proteins with high credibility were obtained after screening. The GO analysis and KEGG analysis was carried out for the 270 proteins to forecast their functions and pathways. Conclusion: The SKP2-binding proteins were screened suc- cessfully, and it was the foundation for the subsequent screening of target-binding proteins and the search for targeting drugs.
Objective： ：The co-immunoprecipitation and mass spectrometric analysis was carried out to obtain the S-phase kinase-asso- ciated protein 2 (SKP2)-binding proteins in HeLa cells, and the biological functions of these binding proteins were forecast. Methods: The co-immunoprecipitation system was established by co-immunoprecipitation and Western blotting assay; the specific protein gel of SKP2-binding proteins was obtained by SDS-PAGE and silver staining assay; the potential SKP2-binding proteins was identified by mass spectrometric analysis; and the GO (Gene ontology) analysis and KEGG analysis was carried out by bioinformatics technique. Results: The expression level of SKP2 protein in HeLa cells was high enough for co-immunoprecipitation assay; the co-immunoprecipi- tation system was established successfully, and SKP2-binding proteins was obtained; a total of 563 proteins were identified by mass spectrometric analysis, and 270 proteins with high credibility were obtained after screening. The GO analysis and KEGG analysis was carried out for the 270 proteins to forecast their functions and pathways. Conclusion: The SKP2-binding proteins were screened suc- cessfully, and it was the foundation for the subsequent screening of target-binding proteins and the search for targeting drugs.
2018, 25(3):263-269.
DOI: 10.3872/j.issn.1007-385X.2018.03.009
Abstract:
Objective: To investigate the impact of FBXW7 gene mutation on cell proliferation, apoptosis, migration, and invasion pro- cesses of colorectal cancer HCT-116 cell line. Methods: Recombinant plasmids carrying wild-type and mutant-type FBXW7 SNP were constructed and transfected into HCT-116 cell line; the FBXW7 protein expression level in HCT-116 strains after transfection was de- tected by Western blotting. Subsequently, cell proliferation capacity was tested by CCK-8 assay; tumor cell colony formation ability was tested by HTCA; cell apoptosis function was tested by FCM; cell migration and invasion were tested by scratch assay and Tran- swell assay, respectively. Results: Higher HBXW7 protein expression level was detected in HCT-116 strain transfected with wild-type HBXW7 in comparison to the control group (strains transfected with mutant-type HBXW7), negative-control (strains transfected with empty plasmids), and blank-control (strains untransfected) (all P<0.05). Compared with the other groups, strains transfected with wild- type HBXW7 exhibited significantly reduced proliferation, colony formation, migration and invasion ability (all P<0.05), but obviously increased apoptosis rate (P<0.05). Conclusion： ：FBXW7 gene mutation can down-regulate its protein expression, and further promote the proliferation, migration and invasion as well as inhibit the apoptosis of HCT-116 cells.
Objective: To investigate the impact of FBXW7 gene mutation on cell proliferation, apoptosis, migration, and invasion pro- cesses of colorectal cancer HCT-116 cell line. Methods: Recombinant plasmids carrying wild-type and mutant-type FBXW7 SNP were constructed and transfected into HCT-116 cell line; the FBXW7 protein expression level in HCT-116 strains after transfection was de- tected by Western blotting. Subsequently, cell proliferation capacity was tested by CCK-8 assay; tumor cell colony formation ability was tested by HTCA; cell apoptosis function was tested by FCM; cell migration and invasion were tested by scratch assay and Tran- swell assay, respectively. Results: Higher HBXW7 protein expression level was detected in HCT-116 strain transfected with wild-type HBXW7 in comparison to the control group (strains transfected with mutant-type HBXW7), negative-control (strains transfected with empty plasmids), and blank-control (strains untransfected) (all P<0.05). Compared with the other groups, strains transfected with wild- type HBXW7 exhibited significantly reduced proliferation, colony formation, migration and invasion ability (all P<0.05), but obviously increased apoptosis rate (P<0.05). Conclusion： ：FBXW7 gene mutation can down-regulate its protein expression, and further promote the proliferation, migration and invasion as well as inhibit the apoptosis of HCT-116 cells.
2018, 25(3):270-274.
DOI: 10.3872/j.issn.1007-385X.2018.03.010
Abstract:
Objective: To investigate the effect of Tim-4 on invasion and migration of cervical cancer cells and its underlying mechanisms. Methods：The expression levels of Tim-4 in cervical cancer cell lines Siha, Hela and cervical epithelial immortalized cell line H8 were detected by Real-time PCR. The Tim-4 lentiviral vector was transfected into Siha cell line. The over-expression of Tim-4 in Siha cell line was detected by fluorescence microscopy. The effects of Tim-4 on the invasion and migration of cervical cancer cell line were detected by Transwell and scratches methods. The changes of MMP2, MMP9, E-cadherin and N-cadherin in Siha cells were detected by Western blotting. Results：The expression of Tim-4 was higher in Siha and Hela cell lines compared to that in the H8 cell line. The Siha cell line burdening Tim-4 lentiviral vector was successfully constructed. Over-expression of Tim-4 significantly inhibited the migration and invasion of cervical cancer cell line, and affected the expression of MMP2, MMP9, N-cadherin and E-cadherin. Conclusion：Over-expression of Tim-4 promotes the invasion and migration by regulating the EMT transformation in cervical cell carcinoma.
Objective: To investigate the effect of Tim-4 on invasion and migration of cervical cancer cells and its underlying mechanisms. Methods：The expression levels of Tim-4 in cervical cancer cell lines Siha, Hela and cervical epithelial immortalized cell line H8 were detected by Real-time PCR. The Tim-4 lentiviral vector was transfected into Siha cell line. The over-expression of Tim-4 in Siha cell line was detected by fluorescence microscopy. The effects of Tim-4 on the invasion and migration of cervical cancer cell line were detected by Transwell and scratches methods. The changes of MMP2, MMP9, E-cadherin and N-cadherin in Siha cells were detected by Western blotting. Results：The expression of Tim-4 was higher in Siha and Hela cell lines compared to that in the H8 cell line. The Siha cell line burdening Tim-4 lentiviral vector was successfully constructed. Over-expression of Tim-4 significantly inhibited the migration and invasion of cervical cancer cell line, and affected the expression of MMP2, MMP9, N-cadherin and E-cadherin. Conclusion：Over-expression of Tim-4 promotes the invasion and migration by regulating the EMT transformation in cervical cell carcinoma.
2018, 25(3):275-280.
DOI: 10.3872/j.issn.1007-385X.2018.03.011
Abstract:
Objective: To compare the differences in clinical and pathological features and survival time between patients with left -sided colon cancer and rectal cancer. Methods: A total of 323 patients with colorectal cancer (CRC) underwent surgical resection at Changhai Hospital of the Second Military Medical University between January 2011 and January 2012 were enrolled in this study. The clinical data of patients were collected and the follow-up was started from the day of surgery or pathological confirmation with the death of patients as endpoints. The follow-up lasted untilAugust 1,2017. Results: There were significant differences in initial symptoms, pathologic type, tumor stage, anemia before surgery, p53 positive rate, and BRAF mutation (χ 2= 59.088，4.188，24.305，11.956，4.221，4.001，all P<0.05) between patients with left-sided colon cancer and rectal cancer. For all the patients, the median survival time was not observed. The five-year survival rates of patients with left-sided colon cancer and rectal cancer were 79.2% and 74.3%, respectively. The Kaplan-Meier survival curves of patients at StageⅠ-Ⅱshowed that there was no statistical difference between patients with left-sided colon cancer and rectal cancer （P=0.840） and the survival of Stage Ⅲ patients between the two groups also showed no statistical difference (P=0.106). Cox regression analysis showed that both the pathologic types [HR=1.759, P=0.047] and tumor stage [HR=2.104, P<0.001] were independent predictive factors for OS of CRC patients. Conclusion: There were no differences in survival time between patients with left-sided colon cancer and rectal cancer. The pathologic types and tumor stage were factors influencing the OS of CRC patients.
Objective: To compare the differences in clinical and pathological features and survival time between patients with left -sided colon cancer and rectal cancer. Methods: A total of 323 patients with colorectal cancer (CRC) underwent surgical resection at Changhai Hospital of the Second Military Medical University between January 2011 and January 2012 were enrolled in this study. The clinical data of patients were collected and the follow-up was started from the day of surgery or pathological confirmation with the death of patients as endpoints. The follow-up lasted untilAugust 1,2017. Results: There were significant differences in initial symptoms, pathologic type, tumor stage, anemia before surgery, p53 positive rate, and BRAF mutation (χ 2= 59.088，4.188，24.305，11.956，4.221，4.001，all P<0.05) between patients with left-sided colon cancer and rectal cancer. For all the patients, the median survival time was not observed. The five-year survival rates of patients with left-sided colon cancer and rectal cancer were 79.2% and 74.3%, respectively. The Kaplan-Meier survival curves of patients at StageⅠ-Ⅱshowed that there was no statistical difference between patients with left-sided colon cancer and rectal cancer （P=0.840） and the survival of Stage Ⅲ patients between the two groups also showed no statistical difference (P=0.106). Cox regression analysis showed that both the pathologic types [HR=1.759, P=0.047] and tumor stage [HR=2.104, P<0.001] were independent predictive factors for OS of CRC patients. Conclusion: There were no differences in survival time between patients with left-sided colon cancer and rectal cancer. The pathologic types and tumor stage were factors influencing the OS of CRC patients.
2018, 25(3):281-287.
DOI: 10.3872/j.issn.1007-385X.2018.03.012
Abstract:
To investigate whether the proliferation and cytotoxicity of NK-92MI cells can be improved by IL-7 and IL-21 genes modification, and determine the effects of this genetically modified NK-92MI cells on T cells from normal human peripheral blood. Methods：IL-7 and IL-21 gene fragments were constructed into electroporation vector by genetic engineering method, and NK- 92MI/IL-21 and NK-92MI/IL-7&21 cells were constructed by electroporation transfection. The in vitro proliferation and cytotoxicity of NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells were measured by cell count and flow cytometry assays. Then, normal human PBMCs were co-cultured with NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells in vitro respectively, and the phenotype change of T cells was measured by flow cytometry. In addition, the cytotoxicity between the activated T cells and three NK-92MI cell lines (NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells) as well as the cytotoxicity of the three NK-92MI cells on tumor cells after co-incubation with activated T cells were detected. Results: NK-92MI/IL-21 cell line (highly expressing IL-21) and NK-92MI/IL-7& 21 cell line (highly expressing both IL-7 and IL-21) were successfully constructed. The toxicity of NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells on Jurkat and K562 cells showed no difference, while the proliferation of NK-92MI/IL-21 and NK-92MI/IL-7&21 cells was increased compared with NK-92MI cells. Furthermore, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells promoted the activation of T cells to a certain degree, and the activated T cells showed merely no cytotoxicity on NK-92MI, NK-92MI/IL-21 and NK-92MI/IL- 7&21 cells; Meanwhile, the activated T cells did not affect the cytotoxicity of the three NK cells (NK-92MI, NK-92MI/IL-21, and NK- 92MI/IL-7&21 cells) on K562 cells under their co-existence. Conclusion: The in vitro proliferation of NK-92MI/IL-21 and NK-92MI/ IL-7&21 cells were enhanced after gene modification, which could also stimulate and promote the activation of T cells from peripheral blood. The cytotoxicity assay showed that the activated T cells had no cytotoxicity on NK-92MI, NK-92MI/IL-21, and NK-92MI/IL-7& 21 cells. Meanwhile, the presence of the activated T cells did not affect the cytotoxicity of NK-92MI cells.
To investigate whether the proliferation and cytotoxicity of NK-92MI cells can be improved by IL-7 and IL-21 genes modification, and determine the effects of this genetically modified NK-92MI cells on T cells from normal human peripheral blood. Methods：IL-7 and IL-21 gene fragments were constructed into electroporation vector by genetic engineering method, and NK- 92MI/IL-21 and NK-92MI/IL-7&21 cells were constructed by electroporation transfection. The in vitro proliferation and cytotoxicity of NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells were measured by cell count and flow cytometry assays. Then, normal human PBMCs were co-cultured with NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells in vitro respectively, and the phenotype change of T cells was measured by flow cytometry. In addition, the cytotoxicity between the activated T cells and three NK-92MI cell lines (NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells) as well as the cytotoxicity of the three NK-92MI cells on tumor cells after co-incubation with activated T cells were detected. Results: NK-92MI/IL-21 cell line (highly expressing IL-21) and NK-92MI/IL-7& 21 cell line (highly expressing both IL-7 and IL-21) were successfully constructed. The toxicity of NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells on Jurkat and K562 cells showed no difference, while the proliferation of NK-92MI/IL-21 and NK-92MI/IL-7&21 cells was increased compared with NK-92MI cells. Furthermore, NK-92MI/IL-21 and NK-92MI/IL-7&21 cells promoted the activation of T cells to a certain degree, and the activated T cells showed merely no cytotoxicity on NK-92MI, NK-92MI/IL-21 and NK-92MI/IL- 7&21 cells; Meanwhile, the activated T cells did not affect the cytotoxicity of the three NK cells (NK-92MI, NK-92MI/IL-21, and NK- 92MI/IL-7&21 cells) on K562 cells under their co-existence. Conclusion: The in vitro proliferation of NK-92MI/IL-21 and NK-92MI/ IL-7&21 cells were enhanced after gene modification, which could also stimulate and promote the activation of T cells from peripheral blood. The cytotoxicity assay showed that the activated T cells had no cytotoxicity on NK-92MI, NK-92MI/IL-21, and NK-92MI/IL-7& 21 cells. Meanwhile, the presence of the activated T cells did not affect the cytotoxicity of NK-92MI cells.
2018, 25(3):288-292.
DOI: 10.3872/j.issn.1007-385X.2018.03.013
Abstract:
胰腺癌是恶性程度最高的消化系统肿瘤，免疫治疗在胰腺癌领域的研究取得较大进展。目前对免疫检查点的研究主 要集中在细胞毒T淋巴细胞抗原4(cytotoxic T lymphocyte antigen-4,CTLA-4)及程序性细胞死亡分子1 (programmed death-1，PD- 1)/程序性死亡蛋白配体1 (programmed death-ligand 1,PD-L1)等分子的研究。已有大量疫苗应用于胰腺癌：靶向KRas、MUC-1/ CEA、WT1 (Wilms tumor-1) 、热激蛋白、多肽疫苗以及VEGFR2 等，其中取得较好效果的有全肿瘤疫苗(如algenpantucel-L)、端粒 酶多肽疫苗(GV1001)、GVAX瘤苗和WT1疫苗等。T细胞也可以控制胰腺癌的进展，大多数免疫疗法在胰腺癌的临床前期实验 中依靠改进T细胞功能来提高疗效，但未来应用于临床还有待进一步深入研究。
胰腺癌是恶性程度最高的消化系统肿瘤，免疫治疗在胰腺癌领域的研究取得较大进展。目前对免疫检查点的研究主 要集中在细胞毒T淋巴细胞抗原4(cytotoxic T lymphocyte antigen-4,CTLA-4)及程序性细胞死亡分子1 (programmed death-1，PD- 1)/程序性死亡蛋白配体1 (programmed death-ligand 1,PD-L1)等分子的研究。已有大量疫苗应用于胰腺癌：靶向KRas、MUC-1/ CEA、WT1 (Wilms tumor-1) 、热激蛋白、多肽疫苗以及VEGFR2 等，其中取得较好效果的有全肿瘤疫苗(如algenpantucel-L)、端粒 酶多肽疫苗(GV1001)、GVAX瘤苗和WT1疫苗等。T细胞也可以控制胰腺癌的进展，大多数免疫疗法在胰腺癌的临床前期实验 中依靠改进T细胞功能来提高疗效，但未来应用于临床还有待进一步深入研究。
2018, 25(3):293-299.
DOI: 10.3872/j.issn.1007-385X.2018.03.014
Abstract:
人体免疫系统是由固有免疫和适应性免疫应答组成。适应性免疫应答在抗原入侵时扮演着至关重要的角色，而CD4 + 辅助性T（CD4 + T helper, CD4 + Th）细胞是适应性免疫应答的主要组成部分。最近新发现的一类不同于Th1和Th2细胞亚群且能 特征性分泌白细胞介素17 (interleukin-17, IL-17)的辅助性T细胞亚群，被命名为Th17细胞。Th17 细胞参与很多炎症性疾病、自 身免疫性疾病和肿瘤等的发展过程，可通过分泌IL-17A、IL-17F、IL-21、IL-22、IL-23、粒细胞-巨噬细胞克隆刺激因子(granulocyte- macrophage colony stimulating factor，GM-CSF)和干扰素γ （interferon-gamma，IFN-γ）等炎症细胞因子来发挥免疫效应和炎症效 应。但是Th17细胞是否参与肿瘤的发生发展、具体作用机制以及发挥促肿瘤还是抑肿瘤效应等问题存在很多争议。本文综述 了近年来Th17细胞分化调节过程的相关机制，以及其在肿瘤发生发展的作用，旨在为肿瘤的诊断和治疗提供新的思路。
人体免疫系统是由固有免疫和适应性免疫应答组成。适应性免疫应答在抗原入侵时扮演着至关重要的角色，而CD4 + 辅助性T（CD4 + T helper, CD4 + Th）细胞是适应性免疫应答的主要组成部分。最近新发现的一类不同于Th1和Th2细胞亚群且能 特征性分泌白细胞介素17 (interleukin-17, IL-17)的辅助性T细胞亚群，被命名为Th17细胞。Th17 细胞参与很多炎症性疾病、自 身免疫性疾病和肿瘤等的发展过程，可通过分泌IL-17A、IL-17F、IL-21、IL-22、IL-23、粒细胞-巨噬细胞克隆刺激因子(granulocyte- macrophage colony stimulating factor，GM-CSF)和干扰素γ （interferon-gamma，IFN-γ）等炎症细胞因子来发挥免疫效应和炎症效 应。但是Th17细胞是否参与肿瘤的发生发展、具体作用机制以及发挥促肿瘤还是抑肿瘤效应等问题存在很多争议。本文综述 了近年来Th17细胞分化调节过程的相关机制，以及其在肿瘤发生发展的作用，旨在为肿瘤的诊断和治疗提供新的思路。
2018, 25(3):300-304.
DOI: 10.3872/j.issn.1007-385X.2018.03.015
Abstract:
黑色素瘤缺乏因子2(absent in melanoma 2, AIM2)定位于细胞质中，可作为模式识别受体感受释放到胞质中的dsD- NA，通过与ASC接头蛋白结合形成炎性小体，从而激活Caspase-1促进炎性细胞因子的分泌和成熟，启动固有免疫应答或细胞焦 亡(pyroptosis)。AIM2被认为是一种肿瘤抑制因子，在多种肿瘤中有异常表达，在肿瘤的发生发展中起到重要作用，能调控急性 电离辐射和化疗引发的结直肠炎症。炎性小体在维持肠道内环境稳态的过程中也起到重要的作用。AIM2能调控细胞周期，抑 制细胞异常增殖，PIK3/Akt通路在结直肠癌（colorectal cancer, CRC)的发生发展中起到重要作用。AIM2能促使IFN-γ和IL-1β分 泌，激发抗肿瘤免疫应答。AIM2还能调控肠道干细胞扩增，调节肠道菌群。AIM2的缺失与CRC的不良预后显著相关，其表达 在CRC的发生发展中起到重要的预后价值。研究AIM2炎性小体在抗肿瘤免疫应答中的作用，对探索和优化CRC免疫治疗过程 的方法有着重要的意义。
黑色素瘤缺乏因子2(absent in melanoma 2, AIM2)定位于细胞质中，可作为模式识别受体感受释放到胞质中的dsD- NA，通过与ASC接头蛋白结合形成炎性小体，从而激活Caspase-1促进炎性细胞因子的分泌和成熟，启动固有免疫应答或细胞焦 亡(pyroptosis)。AIM2被认为是一种肿瘤抑制因子，在多种肿瘤中有异常表达，在肿瘤的发生发展中起到重要作用，能调控急性 电离辐射和化疗引发的结直肠炎症。炎性小体在维持肠道内环境稳态的过程中也起到重要的作用。AIM2能调控细胞周期，抑 制细胞异常增殖，PIK3/Akt通路在结直肠癌（colorectal cancer, CRC)的发生发展中起到重要作用。AIM2能促使IFN-γ和IL-1β分 泌，激发抗肿瘤免疫应答。AIM2还能调控肠道干细胞扩增，调节肠道菌群。AIM2的缺失与CRC的不良预后显著相关，其表达 在CRC的发生发展中起到重要的预后价值。研究AIM2炎性小体在抗肿瘤免疫应答中的作用，对探索和优化CRC免疫治疗过程 的方法有着重要的意义。
2018, 25(3):305-309.
DOI: 10.3872/j.issn.1007-385X.2018.03.016
Abstract:
TET1 （ten-eleven-translocation 1）是一种羟甲基化酶基因，该酶能够催化5甲基胞嘧啶（5-methyl-cytosine，5mC）形成5 羟甲基胞嘧啶（5-hydroxymethyl-cytosine，5hmC），在DNA甲基化调控中发挥重要作用。最近研究表明， TET1 的低表达与肿瘤发 生有关，可作为癌症治疗潜在标志物，表明 TET1 可作为肿瘤抑制基因。此外，除了它的双氧酶活性外， TET1 还可以诱导上皮细 胞间质转变，并充当调节基因转录的辅助活化因子，如癌症的低氧应答基因的调节因子。这表明 TET1 也可做为癌基因促进肿瘤 的发生。因此，在癌症和发育生物学中，TET1的调控机制是十分复杂的， TET1 基因突变也已有报道。本文就TET1在肿瘤发生中 的不同作用进行了综述，深入阐述TET1的作用对拓展DNA去甲基化机制的认识及发现肿瘤治疗新靶标具有重要价值。
TET1 （ten-eleven-translocation 1）是一种羟甲基化酶基因，该酶能够催化5甲基胞嘧啶（5-methyl-cytosine，5mC）形成5 羟甲基胞嘧啶（5-hydroxymethyl-cytosine，5hmC），在DNA甲基化调控中发挥重要作用。最近研究表明， TET1 的低表达与肿瘤发 生有关，可作为癌症治疗潜在标志物，表明 TET1 可作为肿瘤抑制基因。此外，除了它的双氧酶活性外， TET1 还可以诱导上皮细 胞间质转变，并充当调节基因转录的辅助活化因子，如癌症的低氧应答基因的调节因子。这表明 TET1 也可做为癌基因促进肿瘤 的发生。因此，在癌症和发育生物学中，TET1的调控机制是十分复杂的， TET1 基因突变也已有报道。本文就TET1在肿瘤发生中 的不同作用进行了综述，深入阐述TET1的作用对拓展DNA去甲基化机制的认识及发现肿瘤治疗新靶标具有重要价值。
2018, 25(3):310-314.
DOI: 10.3872/j.issn.1007-385X.2018.03.017
Abstract:
P54nrb /NonO是一种能够行使多种生物学功能的核蛋白，在人体多种组织和细胞系中广泛表达并具有高度的种间保守 性，与多种疾病的发生、发展密切相关。近年来，P54 nrb /NonO在多种肿瘤中的作用报道逐渐增多，然而其具体的肿瘤生物学作用 和分子机制尚不明确。P54 nrb /NonO在乳腺癌、前列腺癌、结直肠癌、黑色素瘤、鼻咽癌、肺癌、血液系统肿瘤等多种肿瘤中发挥重 要作用。
P54nrb /NonO是一种能够行使多种生物学功能的核蛋白，在人体多种组织和细胞系中广泛表达并具有高度的种间保守 性，与多种疾病的发生、发展密切相关。近年来，P54 nrb /NonO在多种肿瘤中的作用报道逐渐增多，然而其具体的肿瘤生物学作用 和分子机制尚不明确。P54 nrb /NonO在乳腺癌、前列腺癌、结直肠癌、黑色素瘤、鼻咽癌、肺癌、血液系统肿瘤等多种肿瘤中发挥重 要作用。
2018, 25(3):315-317.
DOI: 10.3872/j.issn.1007-385X.2018.03.018
Abstract:
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
- 电话：021-81871002-22
- 电子邮箱：cjcb@biother.cn
- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
- 国内定价： ￥20元/册
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.