摘要
目的 生存素 (survivin)是近年来发现的凋亡抑制蛋白家族新成员 ,在多数恶性肿瘤组织中丰富表达。因此 ,观察生存素反义寡核苷酸转染对肝癌细胞凋亡、增殖、细胞对化疗药物敏感性的影响。方法 设计合成特异性靶向生存素的反义寡核苷酸 (ASODN)。肝癌细胞株hepG2 分为 6组 :空白对照组、脂质体转染对照组、正义链转染对照组、2 0 0、40 0和 6 0 0nmol/LASODN转染组。作用 2 0h后收获各组细胞。倒置显微镜观察细胞形态变化 ,Westernblot法检测各组细胞生存素表达情况 ,流式细胞术检测各组细胞增殖和凋亡指数 ,MTT法检测 5 氟尿嘧啶 (5 FU)和顺铂 (DDP)对各组细胞的生长抑制率。结果 各ASODN转染组细胞生存素表达有不同程度减弱 ,细胞变圆、折光增强、漂浮、细胞碎片形成等 ,而各对照组细胞生长良好 ;各ASODN转染组细胞凋亡指数明显高于各对照组 (P <0 .0 5 ) ,以 6 0 0nmol/L转染组最为明显 (P <0 .0 5 ) ,而各对照组间差异无显著性 (P >0 .0 5 ) ;各ASODN转染组细胞增殖指数明显低于各对照组 (P <0 .0 5 ) ,以 6 0 0nmol/L转染组最为明显 (P <0 .0 5 ) ,而各对照组间差异无显著性 (P >0 .0 5 ) ;等浓度化疗药物 5 FU和DDP对各ASODN转染组细胞的抑制率明显高于各对照组(P <0 .0 5 ) ,以 6 0
Objective Survivin is a recently described inhibitor of apoptosis. It is undetectable in normal tissue of adults, but abundantly expressed in transformed cells and a variety of human cancers. This study was aimed to investigate the effect of antisense oligonucleotide (ASODN) targeting survivin on hepatic cell apoptosis, proliferation and sensitivity to chemotherapeutic drugs. Methods ASODN targeting survivin was designed and constructed. Cultured cells were divided into 6 groups: control group, lipofectin group, oligonucleotide(ODN) group, 200 nmol/L ASODN group, 400 nmol/L ASODN group and 600 nmol/L ASODN group. After transfection for 20 h, cultured cells were harvested to carry on the next tests. Cell morphological change was observed with invert microscope; survivin protein expression was detected by Western blot; apoptotic index (AI) and proliferative index (PI) were examined by flow cytometry; rate of inhibition (IR) by chemotherapeutic drugs was determined by the colorimetric MTT cell viability/proliferation assay. Results Expression of survivin in ASODN groups was significantly decreased compared with that in the control, lipofectin and ODN group; abnormal morphological change of cells was observed in ASODN transfected groups; the AI of ASODN groups was significantly higher than that of other groups; the PI of ASODN groups was significantly lower than that of other groups; the IR of chemotherapeutic drugs in ASODN groups was significantly higher than that of other groups. Conclusions Expression of survivin may decrease in hepG2 cells after ASODN transfection. ASODN targeting survivin can induce cell apoptosis, inhibit cell proliferation and sensitize hepatocarcinoma cells to chemotherapy.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2003年第1期11-14,共4页
Chinese Journal of Digestion
