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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 0nmol/L
Objective Survivin is a new member of the apoptosis-inhibitory protein family found in recent years and is abundantly expressed in most malignant tumors. Therefore, we observed the effect of survivin antisense oligonucleotide transfection on hepatocellular carcinoma cell apoptosis, proliferation, cell chemosensitivity. Methods Antisense oligonucleotides (ASODNs) were designed and synthesized to specifically target survivin. Hepatocellular carcinoma cell line hepG2 was divided into 6 groups: blank control group, lipofection control group, sense strand transfected control group, 200, 400, and 600 nmol / L ASODN transfected groups. The cells were harvested after 20 hours. The changes of cell morphology were observed by inverted microscope. The expression of survivin in each group was detected by Western blot. The proliferation and apoptosis index of each group were detected by flow cytometry. The expressions of 5-fluorouracil (5 FU) and cisplatin (DDP) Cell growth inhibition rate. Results The expression of survivin in each ASODN transfection group was weakened to varying degrees, the cells were round, the refraction was enhanced, the floating and the cell debris were formed, while the cells in each control group grew well; the apoptotic index of ASODN transfection group was significantly higher than that of each In control group (P <0.05), the transfection group at 600 nmol / L was the most obvious (P <0.05), while there was no significant difference among the control groups (P> 0.05) The proliferation index of ASODN transfection group was significantly lower than that of each control group (P <0.05), and the most significant difference was in 600 nmol / L transfection group (P <0.05), but there was no significant difference among the control groups (P> 0.05). The inhibitory rates of isoflurane 5 FU and DDP on ASODN transfection group were significantly higher than those of control group (P <0.05)