CAJ | 학술논문

目的构建Livin靶向小干扰RNA(siRNA)重组表达载体,观察其对前列腺癌PC3细胞凋亡和增殖等生物学特性的影响. 方法 2009年1月至2012年1月,构建针对Livin基因的siRNA真核表达载体U6/GFP/Neo-Livin并转染前列腺癌PC3细胞.实时荧光定量PCR和蛋白质印迹法分别检测转染后Livin基因mRNA和蛋白的表达情况,MTT法检测对细胞增殖的影响,流式细胞仪检测转染后细胞的凋亡效应以及对细胞周期的影响,克隆形成实验观察对细胞体外克隆形成能力的改变,体内成瘤实验比较细胞在裸鼠体内成瘤性的变化. 结果 Livin靶向siRNA重组表达载体转染PC3细胞后,与对照组相比,实验组细胞内Livin mRNA和蛋白的表达水平分别下调至(31.55±3.92)％、(0.37±0.02)％(P＜0.01)；与对照组相比,PC3细胞增殖受到显著抑制(P＜0.01)；细胞凋亡率增到(26.5±3.3)％(P＜0.01)；Caspase3活性增至0.079±0.017 (P＜0.05),体外克隆形成率降至(34.8±3.5)％(P＜0.01).实验组和对照组荷瘤裸鼠的成瘤体积分别为(1.79±0.07)、(4.40±0.06) cm3(p＜0.01). 结论靶向Livin基因的siRNA干扰了前列腺癌PC3细胞中Livin基因的表达,抑制了细胞增殖并诱导其凋亡,延缓了肿瘤细胞的生长.
목적 구건Livin파향소간우RNA(siRNA)중조표체재체,관찰기대전렬선암PC3세포조망화증식등생물학특성적영향. 방법 2009년1월지2012년1월,구건침대Livin기인적siRNA진핵표체재체U6/GFP/Neo-Livin병전염전렬선암PC3세포.실시형광정량PCR화단백질인적법분별검측전염후Livin기인mRNA화단백적표체정황,MTT법검측대세포증식적영향,류식세포의검측전염후세포적조망효응이급대세포주기적영향,극륭형성실험관찰대세포체외극륭형성능력적개변,체내성류실험비교세포재라서체내성류성적변화. 결과 Livin파향siRNA중조표체재체전염PC3세포후,여대조조상비,실험조세포내Livin mRNA화단백적표체수평분별하조지(31.55±3.92)％、(0.37±0.02)％(P＜0.01)；여대조조상비,PC3세포증식수도현저억제(P＜0.01)；세포조망솔증도(26.5±3.3)％(P＜0.01)；Caspase3활성증지0.079±0.017 (P＜0.05),체외극륭형성솔강지(34.8±3.5)％(P＜0.01).실험조화대조조하류라서적성류체적분별위(1.79±0.07)、(4.40±0.06) cm3(p＜0.01). 결론 파향Livin기인적siRNA간우료전렬선암PC3세포중Livin기인적표체,억제료세포증식병유도기조망,연완료종류세포적생장.
Objective To observe the effect of RNAi targeting Livin gene on biology characteristics such as apoptosis and proliferation in human prostate cancer cells.Methods siRNA expression vector targeting Livin gene was constructed and transfected into human prostate cancer cell line PC3.The expressions of Livin mRNA and protein were detected by real-time PCR and Western-blot,cell apoptosis and cell cycle were assayed by flow cytometry,proliferation and colony formation were detected by MTT and colony formation assay,and the tumor growth in vivo was observed in nude mice.Results After transfection,downregulation of Livin mRNA and protein expression in PC3 cells was observed (P＜0.01).Compared with the control group,the proliferation of cancer cells was inhibited significantly (P＜0.01) and the apoptotic ratio was (26.5±3.3) ％ (P＜0.01).The Caspase3 activity increased obviously (P＜0.05),and the experimental group showed a decreased colony formation rate (P＜0.01).The tumor volume of xenografts in nude mouse in experimental and control group was (1.79± 0.07) and (4.40 ± 0.06) cm3 respectively (P ＜ 0.01).Conclusions The siRNA recombinant expression vector targeting Livin gene was constructed and can knockdown the expression of Livin mRNA and protein.It can inhibit PC3 cell proliferation,induce apoptosis and inhibit tumor growth in vivo.