沉默AFP基因可抑制肝癌HepG2细胞Survivin mRNA的表达

方念; 方紫凌; 张慧卿; 韩小妮; 黄根; 王农荣; 熊建萍; 张焜和

doi:10.3969/j.issn.1000-8179.20131261

沉默AFP基因可抑制肝癌HepG2细胞Survivin mRNA的表达

Survivin mRNA is suppressed in HepG2 cells after AFP gene silencing

摘要

摘要:
目的探讨肝癌HepG2细胞AFP基因沉默对Survivin表达的影响。
方法通过siRNA技术下调肝癌HepG2细胞中AFP的表达，采用ELISA法测定转染前后上清液AFP浓度，MTT法检测转染前后细胞增殖活性，流式细胞术分析细胞凋亡率，RT-PCR检测转染前后细胞Survivin mRNA水平。
结果转染48 h后，实验组上清液中AFP浓度显著下降，肝癌细胞生长活性下降43.1%，凋亡率增加24.3%，HepG2细胞中Survivin mRNA表达降低78.0%。对照组和空白组的上述指标均无明显变化。
结论沉默肝癌HepG2细胞AFP的表达，能有效抑制肝癌细胞生长、促进细胞凋亡，这一作用可能与细胞内Survivin mRNA水平降低有关。

Abstract:
Objective To observe the effects of AFP gene silencing by siRNA on the Survivin mRNA of hepatocellular carcinoma cell line HepG2.
Methods AFP gene expression was downregulated in HepG2 cell by RNAi, and the AFP content in the supernatant was detected by ELISA. Survivin mRNA level was tested by RT-PCR. MTT was applied to evaluate cell proliferation. Flow cytometry was employed to observe cell apoptosis.
Results At 48 h after transfection, AFP expression was almost completely inhibited, cell proliferation activity was decreased by 43.1%, cell apoptosis rate was increased by 24.3%, and the Survivin mRNA expression was reduced to 22.0% in the experimental group. No evident changes were observed in negative control and blank groups.
Conclusion AFP gene silenced by RNAi induces growth inhibition and apoptosis promotion of hepatocellular carcinoma cell line HepG2. This gene may be associated with the suppression of Survivin mRNA.

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