9 Survivin反义寡核苷酸对人肝癌细胞的抑制作用

目的:探讨survivin反义寡核苷酸（ASODN）对人肝癌细胞HepG2的抑制作用。方法:采用免疫组织化学法检测肝细胞癌（HCC）组织中survivin的表达。采用脂质体介导survivin ASODN体外转染人肝癌细胞株HepG2，Western blot检测细胞survivin蛋白的表达，FCM检测细胞的凋亡率，观察细胞在软琼脂中的集落形成能力。建立人肝癌裸鼠皮下移植瘤模型，观察survivin ASODN的体内抑癌作用。 结果:（1）肝癌组织中survivin表达阳性率为75.8％(25/33)，明显高于癌旁组织和正常肝组织(P<0.01);（2）survivin ASODN体外转染可明显下调HepG2细胞survivin蛋白表达，ASODN组HepG2细胞凋亡率明显高于空白对照组和SODN组(P<0.01)，ASODN组HepG2细胞在软琼脂中形成的集落数目明显少于空白对照组和SODN组(P<0.01);（3）ASODN组瘤体生长速度较空白对照组和SODN组明显减慢(P<0.01)，ASODN组瘤体重量较空白对照组和SODN组明显减轻(P<0.05)。结论:Survivin在HCC组织中高表达;survivin ASODN可以诱导HepG2细胞凋亡，在体外实验和动物实验中对HepG2细胞生长都有抑制作用。

The inhibitory effect of survivin antisense oligonucleotide on human hepatocellular carcinoma cells

Objective To explore the inhibitory effect of survivin antisense oligonucleotide(ASODN) on human hepatocellular carcinoma(HCC) cell line HepG2.Methods The expression of survivin protein in HCC tissue was detected immunohistochemically.Survivin ASODN was transfected into HepG2 cell by lipofectamine in vitro.The expression of survivin protein was detected by western-blot.Apoptotic index(AI) and colony formation were examined by flow cytometry and in soft agar respectively.Human HCC model in nude mice was established by subcutaneous injection of HepG2 cells.In vivo effect of survivin ASODN on tumor growth was observed.Results(1) The expression rate of survivin protein in HCC was 75.8 %(25/33),which was significantly higher than that in corresponding non-cancerous adjacent liver tissue and normal liver tissue(P<0.01).(2) The expression of survivin protein in HepG2 cells was obviously downregulated after transfected with survivin ASODN.AI of HepG2 cells in ASODN group were significantly higher than those in the control and SODN groups(P<0.01).Colonies of HepG2 cells in ASODN group were significantly fewer those that in the control and SODN groups(P<0.01).(3) The tumor growth of nude mice in ASODN group was significantly slower than that in the control and SODN groups(P< 0.01).The tumor weight of ASODN group was significantly lower than that of the control and SODN groups(P<0.05).Conclusions There is overexpression of survivin in HCC.Survivin ASODN can induce HepG2 cells apoptosis and inhibit the growth of HepG2 cells in vitro and in vivo.