AIM:To investigate whether silencing Fas-associated phosphatase 1(FAP-1)expression enhances the efficiency of chemotherapy for colon carcinoma with oxaliplatin.METHODS:Expression of FAP-1 in mRNA and protein was detec...AIM:To investigate whether silencing Fas-associated phosphatase 1(FAP-1)expression enhances the efficiency of chemotherapy for colon carcinoma with oxaliplatin.METHODS:Expression of FAP-1 in mRNA and protein was detected by reverse transcription polymerase chain reaction(RT-PCR)and flow cytometry.Small interfering RNA(siRNA)was designed according to the FAP-1 mRNA sequence.Cell proliferation was evaluated by methyl thiazolyl tetrazolium(MTT)assay.Anenxin V-and propidine iodine(PI)were assayed by flow cytometry for the detection of apoptosis. RESULTS:The expression of FAP-1 was increased in SW480 cells after chemotherapy with oxaliplatin. Transfection of FAP-1 siRNA into SW480 cells silenced the expression of FAP-1 and consequently abolished the inhibitory function of Fas/FasL-mediated apoptosis pathway,thus increasing the efficacy of chemotherapy for colon carcinoma with oxaliplatin. CONCLUSION:RNA interference combined with conventional chemotherapy is more effective against colon cancer.展开更多
目的:检测过表达F a s相关蛋白1(F a sassociated factor 1,FAF1)对胃癌细胞HGC-27增殖及凋亡的影响,探讨FAF1与胃癌发生发展的相关性.方法:实验分为阴性对照组(未转染组)、空载转染组(感染空载体慢病毒颗粒1.0×108TU/mL)、过表达F...目的:检测过表达F a s相关蛋白1(F a sassociated factor 1,FAF1)对胃癌细胞HGC-27增殖及凋亡的影响,探讨FAF1与胃癌发生发展的相关性.方法:实验分为阴性对照组(未转染组)、空载转染组(感染空载体慢病毒颗粒1.0×108TU/mL)、过表达FAF1组(感染FAF1过表达慢病毒颗粒1.0×108TU/mL).激光共聚焦显微镜观察转染效率,Western blot检测FAF1蛋白的表达情况,透射电镜观察细胞超微结构变化,流式细胞仪检测细胞周期分布及凋亡情况,MTT检测细胞生长情况.结果:依据GFP绿色荧光可测定细胞转染效率达95%以上;与阴性对照组和空载转染组相比,过表达FAF1组FAF1蛋白明显高表达;阴性对照组和空载转染组均细胞膜完整,细胞核正常,两者超微结构无明显差异,而过表达FAF1组胞核裂解成碎片状,可见凋亡小体形成;FAF1过表达能够抑制人胃癌细胞HGC-27的增殖,诱导细胞凋亡,改变细胞周期的分布,与阴性对照组和空载转染组相比,过表达FAF1组细胞的倍增时间显著延长,细胞凋亡率显著提高(84.66%±5.92%vs 4.60%±3.80%和7.32%±3.82%,P<0.05),G0/G1期细胞显著降低(46.43%±2.43%vs 54.93%±3.5%和54%±0.3%,P<0.05),G2/M期细胞显著增多(29.78%±3.91%vs 19.33%±3.82%和20.93%±2.46%,P<0.05),差异均有统计学意义.结论:构建的过表达FAF1慢病毒可以抑制胃癌细胞的生长,改变细胞周期的分布,促进胃癌细胞的凋亡.展开更多
基金Supported by Research grants from the Science and Technology Foundation of Guangdong Province,No.2006B36002010,2008B030301092,2009B030801005the Foundation of Health Department of Guangdong Province,No.A2005226+2 种基金the foundation of Guangzhou Science and Technology Bureau,No. 2009Y-C011-1the Natural Science Foundation of Guangdong Province,No.7001592the National Natural Science Foundation of China,No.30973505
文摘AIM:To investigate whether silencing Fas-associated phosphatase 1(FAP-1)expression enhances the efficiency of chemotherapy for colon carcinoma with oxaliplatin.METHODS:Expression of FAP-1 in mRNA and protein was detected by reverse transcription polymerase chain reaction(RT-PCR)and flow cytometry.Small interfering RNA(siRNA)was designed according to the FAP-1 mRNA sequence.Cell proliferation was evaluated by methyl thiazolyl tetrazolium(MTT)assay.Anenxin V-and propidine iodine(PI)were assayed by flow cytometry for the detection of apoptosis. RESULTS:The expression of FAP-1 was increased in SW480 cells after chemotherapy with oxaliplatin. Transfection of FAP-1 siRNA into SW480 cells silenced the expression of FAP-1 and consequently abolished the inhibitory function of Fas/FasL-mediated apoptosis pathway,thus increasing the efficacy of chemotherapy for colon carcinoma with oxaliplatin. CONCLUSION:RNA interference combined with conventional chemotherapy is more effective against colon cancer.
