Survivin反义寡核苷酸对人卵巢癌细胞SKOV3的抑制作用

Experimental Therapy of Survivin Antisense Oligonucleotide for Human Ovarian Cancer Cell SKOV3

背景与目的在大多数肿瘤组织中都发现了survivin的异常高表达,这说明survivin在肿瘤发生中具有重要作用。本研究探讨survivin反义寡核苷酸(survivinASODN)对人卵巢癌细胞SKOV3的抑制作用及其机制。方法脂质体介导survivin反义寡核苷酸转染SKOV3,用MTT法检测细胞抑制率。Westernblot法检测相关基因蛋白表达,应用流式细胞术、梯形DNA片段分析及DAPI染色光镜观察细胞凋亡情况。激酶活性检测方法测定半胱氨蛋白水解酶3(caspase-3)活性的变化。结果脂质体介导survivin反义寡核苷酸转染后,SKOV3细胞的生长受到明显抑制,1000ng/mlASODN转染组的细胞抑制率为(60.30±2.95)%,明显高于对照组(P<0.05)。凋亡细胞比率由转染前的0.65%增加至32.10%,SKOV3细胞内survivin基因蛋白表达下调26.3%,caspase-3活性为0.998±0.001,较对照组显著增高(P<0.01)。结论SurvivinASODN可通过诱导细胞凋亡抑制细胞生长,具有明显的抗癌作用。

BACKGROUND &OBJECTIVE:Survivin abnormally overexpresses in a variety of human tumors, it may play an important role in the development of tumor. This study was designed to investigate the inhibitory effects of survivin antisense oligonucleotide (ASODN) on human ovarian carcinoma cell SKOV3 in vivo and its mechanism. METHODS: SKOV3 cells were transfected with survivin ASODN mediated by DOTAP liposome reagent, MTT method was used to observe the inhibitory rate, Western blot analysis was used to determine relating gene expression. Flow cytometry, DNA ladder analysis, and DAPI staining were used to examine cell apoptosis. Kinase activity test was used to examine the changes of caspase 3 activity. RESULTS: The expression of survivin in SKOV3 cells decreased after transfected with survivin ASODN. Survivin ASODN has obviously inhibited the growth of SKOV3 cells after transfection. The inhibitory rate of 1 000 ng/ml ASODN transfection group was (60.30±2.95)%, which is obviously higher than control group (P< 0.05). Survivin ASODN transfection induced cell cycle arrest in G1 phrase, while the apoptotic rate increased from 0.65%to 32.10%. Caspase 3 activity (0.998±0.001) increased significantly after transfected with survivin ASODN (P< 0.01). CONCLUSION: Survivin ASODN,which can inhibit human ovarian carcinoma SKOV3 cells proliferation and induce apoptosis, is a prospective anti cancer drug.