siRNA沉默ENAH基因对肝癌细胞生长的抑制作用

Inhibition of ENAH gene silenced by siRNA on growth of liver cancer cell

目的探讨siRNA沉默ENAH基因对肝癌细胞生长的抑制作用。方法采用脂质体Lipofectamine 2000将ENAH-siRNA及control-siRNA转染至人肝癌细胞HCCLM3,分别设为ENAHsiRNA组和control-siRNA组。采用Western blot检测ENAH蛋白表达,采用裸鼠成瘤实验观察肝癌细胞致瘤能力。记录肿瘤体积,绘制生长曲线,进行生存分析。两组实验数据比较采用t检验,生存分析采用Kaplan-Meier法和Log-rank检验。结果 ENAH-siRNA组细胞ENAH蛋白表达明显弱于controlsiRNA组。ENAH-siRNA组裸鼠接种后实体肿瘤形成时间为（24±3）d,明显长于control-siRNA组的（8±2）d（t=12.55,P〈0.05）。ENAH-siRNA组裸鼠中位生存时间为64（48~81）d,control-siRNA组为34（21~48）d,两组总体生存率比较差异有统计学意义（χ2=14.33,P〈0.05）。结论 siRNA沉默ENAH基因可明显减弱肝癌细胞的致瘤能力,抑制肿瘤生长。

Objective To investigate the inhibition of ENAH gene silenced by siRNA on the growth of liver cancer cell. Methods ENAH-siRNA and control-siRNA were transfected to human liver cancer cell HCCLM3 using liposome Lipofectamine 2000 and ENAH-siRNA group and control-siRNA group were established. The expression of ENAH protein was detected by Western blot. Tumorigenic ability of the liver cancer cell was observed through nude mice tumorigenicity assay. Tumor volume was recorded, growth curve was drawn and survival analysis was conducted. The experimental data of two groups were compared using t test, and the survival analysis was conducted using Kaplan-Meier method and Log-rank test. Results The expression of ENAH protein in ENAH- siRNA group was significantly less than that in control-siRNA group. The formation time of solid tumor after inoculation in nude mice in ENAH-siRNA group was （24±3） d, which was significantly longer than （8±2） d in control-siRNA group （t=12.55, P〈0.05）. The median survival time of the nude mice was 64 （48 ± 81） d in ENAH-siRNA group and was 34 （21 ± 48） d in control-siRNA group. There was significant difference in the overall survival rate between two groups （x^2=14.33, P〈0.05）. Conclusion ENAH gene silenced by siRNA may obviously weaken the tumorigenic ability of liver cancer cell and inhibit the growth of tumor.