Objective: To investigate the antitumor effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene transfection mediated by adenovirus into human pancreatic carcinoma cell line Panc-1, and the mech...Objective: To investigate the antitumor effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene transfection mediated by adenovirus into human pancreatic carcinoma cell line Panc-1, and the mechanisms involved in this effect. Methods: TRAIL gene was transfected into pancreatic cancer cell line Panc-1 by an adenovirus vector (Ad-TRAIL). Level of TRAIL mRNA expression was determined using RT-PCR, and TRAIL protein synthesis was evaluated with Western blot. Cell-growth activities were determined by MTT assay. The bystander effect was observed by co-culturing the Panc-1 cells with the transfected TRAIL gene at different ratios. Apoptosis in pancreatic cancer cells was detected by flow cytometry. Procaspase-8 and procaspase-3 were determined by Western blot. Results: The stable overexpression of TRAIL was de-tected in Panc-1 cells transfected by Ad-TRAIL. Ad-TRAIL significantly inhibited of cell viability of Panc-1 cells. Furthermore, co-culture of cancer cells transfected with TRAIL with that nontransfected resulted in the cell death of both cells by bystander effect. Moreover, the percentage of apoptotic cells was significantly higher in the Ad-TRAIL-treatment group compared to the control groups (P < 0.01). And there was a diminished amount of procaspase-8 and procaspase-3 after infection with Ad-TRAIL. Conclusion: The overexpression of TRAIL gene in Panc-1 cells by Ad-TRAIL exerts its antitumor effects, and the mechanisms involved in this effect may be proapoptosis and bystander effect.展开更多
Objective Gemcitabine, the only approved drug for the treatment of pancreatic cancer, is not very effective. Novel and effective cancer chemopreventive agents are urgently needed. Recently, emerging studies determined...Objective Gemcitabine, the only approved drug for the treatment of pancreatic cancer, is not very effective. Novel and effective cancer chemopreventive agents are urgently needed. Recently, emerging studies determined resveratrol possessed anticancer effects on various cancer cells. We explored the anticancer effect of resveratrol in pancreatic cancer cells and investigated the involved moleculars of action. We also examined whether resveratrol enhanced antitumor activity of gemcitabine in vitro.Methods Proliferation inhibition was assessed by cell count kit-8 assay. Cell cycle phase distribution and apoptotic cells were measured by flow cytometric analysis. We determined the expression of bcl-2, cyclinD1, and activation of caspases-3 and poly(ADP-ribose) polymerase1 proteins used Western blot analysis.Results Resveratrol inhibited the proliferation of three pancreatic cancer cell lines in a dose dependent fashion, and induced accumulation of cells at the G1 phase as well as apoptosis. Our data also demonstrated that resveratrol enhanced gemcitabine-induced apoptosis in pancreatic cancer cells. In addition, resveratrol inhibited the expression of cyclinD1, bcl-2, and induced activation of caspase-3 and poly(ADPribose) polymerase1. Conclusion Our results suggested that resveratrol might be not only a potential regimen, but also an effective chemosensitizer for the chemotherapy of pancreatic cancer.展开更多
基金Supported by a grant from the National Natural Science Foundation of China (No. 30471693).
文摘Objective: To investigate the antitumor effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene transfection mediated by adenovirus into human pancreatic carcinoma cell line Panc-1, and the mechanisms involved in this effect. Methods: TRAIL gene was transfected into pancreatic cancer cell line Panc-1 by an adenovirus vector (Ad-TRAIL). Level of TRAIL mRNA expression was determined using RT-PCR, and TRAIL protein synthesis was evaluated with Western blot. Cell-growth activities were determined by MTT assay. The bystander effect was observed by co-culturing the Panc-1 cells with the transfected TRAIL gene at different ratios. Apoptosis in pancreatic cancer cells was detected by flow cytometry. Procaspase-8 and procaspase-3 were determined by Western blot. Results: The stable overexpression of TRAIL was de-tected in Panc-1 cells transfected by Ad-TRAIL. Ad-TRAIL significantly inhibited of cell viability of Panc-1 cells. Furthermore, co-culture of cancer cells transfected with TRAIL with that nontransfected resulted in the cell death of both cells by bystander effect. Moreover, the percentage of apoptotic cells was significantly higher in the Ad-TRAIL-treatment group compared to the control groups (P < 0.01). And there was a diminished amount of procaspase-8 and procaspase-3 after infection with Ad-TRAIL. Conclusion: The overexpression of TRAIL gene in Panc-1 cells by Ad-TRAIL exerts its antitumor effects, and the mechanisms involved in this effect may be proapoptosis and bystander effect.
基金Supported by grants from the Eight Natural Science Foundation(No.81272611)Major Science and Technology Innovation Project of Hangzhou(No.20112312A01),China
文摘Objective Gemcitabine, the only approved drug for the treatment of pancreatic cancer, is not very effective. Novel and effective cancer chemopreventive agents are urgently needed. Recently, emerging studies determined resveratrol possessed anticancer effects on various cancer cells. We explored the anticancer effect of resveratrol in pancreatic cancer cells and investigated the involved moleculars of action. We also examined whether resveratrol enhanced antitumor activity of gemcitabine in vitro.Methods Proliferation inhibition was assessed by cell count kit-8 assay. Cell cycle phase distribution and apoptotic cells were measured by flow cytometric analysis. We determined the expression of bcl-2, cyclinD1, and activation of caspases-3 and poly(ADP-ribose) polymerase1 proteins used Western blot analysis.Results Resveratrol inhibited the proliferation of three pancreatic cancer cell lines in a dose dependent fashion, and induced accumulation of cells at the G1 phase as well as apoptosis. Our data also demonstrated that resveratrol enhanced gemcitabine-induced apoptosis in pancreatic cancer cells. In addition, resveratrol inhibited the expression of cyclinD1, bcl-2, and induced activation of caspase-3 and poly(ADPribose) polymerase1. Conclusion Our results suggested that resveratrol might be not only a potential regimen, but also an effective chemosensitizer for the chemotherapy of pancreatic cancer.
