CAJ | 학술논문

目的探讨以Survivin为靶基因的胰腺癌基因治疗的可能性,为胰腺癌基因治疗提供依据.方法采用化学合成的小十扰RNA(siRNA)和pGCSi载体中的小发夹RNA(shRNA)抑制胰腺癌细胞系PaTu8988的Sunrivin基因表达,通过观察胰腺癌细胞株Survivin基因表达的下调以及细胞形态、细胞凋亡、细胞活力、凋亡信号途径等的改变评价Survivin作为靶基因的治疗效果.结果不同序列的siRNA和shRNA抑制胰腺癌细胞Survivin的表达后,Survivin mRNA和蛋白表达水平明显下降(P<0.05);碘化吡啶(PI)染色法观察发现RNA干扰(RNAi)后细胞出现核皱缩、细胞凋亡率>20%;流式细胞仪检测发现RNAi后在G0/G1期前出现了亚二倍体峰(P<0.05);Western blot法检测发现RNAi后Casptrqe-3被激活(P<0.05).结论通过抑制胰腺癌细胞PaTu8988的Survivin表达可以诱导肿瘤细胞启动凋亡程序,加速肿瘤细胞凋亡,由此可望提高胰腺癌的治疗效果.
목적 탐토이Survivin위파기인적이선암기인치료적가능성,위이선암기인치료제공의거.방법 채용화학합성적소십우RNA(siRNA)화pGCSi재체중적소발협RNA(shRNA)억제이선암세포계PaTu8988적Sunrivin기인표체,통과관찰이선암세포주Survivin기인표체적하조이급세포형태、세포조망、세포활력、조망신호도경등적개변평개Survivin작위파기인적치료효과.결과 불동서렬적siRNA화shRNA억제이선암세포Survivin적표체후,Survivin mRNA화단백표체수평명현하강(P<0.05);전화필정(PI)염색법관찰발현RNA간우(RNAi)후세포출현핵추축、세포조망솔>20%;류식세포의검측발현RNAi후재G0/G1기전출현료아이배체봉(P<0.05);Western blot법검측발현RNAi후Casptrqe-3피격활(P<0.05).결론 통과억제이선암세포PaTu8988적Survivin표체가이유도종류세포계동조망정서,가속종류세포조망,유차가망제고이선암적치료효과.
Objective To investigate the influence of gene therapy using survivin as a gene target on biological behavior of pancreatic carcinoma cell line. Methods Chemically synthesized siRNA and shRNA in pGCSi vector were used to silence survivin expression of pancreatic carcinoma cell line PaTu8988. The therapeutical effects of survivin as a gene target were evaluated through determination of the down-regulation of survivin gene expression, cellular shape, cell apoptosis, cell viability and apoptosis signal pathway changes. Results After transfection of different arrays of siRNA and shRNA vectors to silence the survivin expression, survivin mRNA and protein levels were significantly decreased (P < 0.05) ; PI staining revealed the presence of karyopyknosis, the cell apoptosis index was more than 20%; hypodiploid DNA content before G0/G1 detected by flow cytometry ; cell viability measured by MTT assay was significantly decreased (P <0.05) ; the activity of caspase-3 remarkably increased (P < 0. 05). Conclusions The pancreatic carcinoma cell line PaTu8988 be induced to promote spontaneous apoptosis procedure through silencing survivin expression by RNAi, which could accelerate carcinoma cell apoptosis and improve therapeutic effect on pancreatic carcinoma.