南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (08): 875-.doi: 10.12122/j.issn.1673-4254.2019.08.01

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E26特异性转化变异体4在体外促进肝癌细胞对索拉非尼和顺铂耐药

陈筱卉，李欣，吴德华   

  • 出版日期:2019-08-20 发布日期:2019-08-20

E26 transformation-specific variant 4 promotes sorafenib and cisplatin resistance in hepatocellular carcinoma cells in vitro

  • Online:2019-08-20 Published:2019-08-20

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摘要： 目的研究E26特异性转化变异体4（ETV4）对肝细胞癌（HCC）顺铂和索拉非尼耐药的影响。方法顺铂耐药HCC细胞株 SMMC-7721（对索拉非尼敏感）和HCC-LM3（对索拉非尼不敏感）经质粒转染诱导ETV4过表达或用siRNA干扰ETV4后，分别 用DMSO、索拉非尼（5 μmol/L）或顺铂（5 μmol/L）刺激48 h并收集总蛋白或总RNA。采用Western blotting、流式细胞术、EdU增 殖检测法检测处理后HCC细胞凋亡水平和增殖能力的变化。利用实时荧光定量PCR（q-PCR）技术检测11例HCC患者肿瘤组 织及配对的癌旁组织中ETV4 mRNA的表达水平。肝癌细胞株经干扰ETV4后，分别用DMSO、索拉非尼或顺铂刺激48 h，利用 q-RCR技术检测早期反应基因3（IER3）的mRNA表达水平。结果肝癌组织中ETV4 mRNA的表达水平显著高于对应的癌旁 组织。在两株肝癌细胞中过表达ETV4能显著减少索拉非尼或顺铂诱导的细胞凋亡，而干扰ETV4能显著增加索拉非尼或顺铂 诱导的细胞凋亡，并显著抑制索拉非尼或顺铂刺激下的细胞增殖能力。此外，在索拉非尼或顺铂的刺激下ETV4能调节IER3的 mRNA表达水平。结论ETV4过表达能促进HCC细胞对索拉非尼或顺铂产生耐药。

Abstract: Objective To investigate the role of E26 transformation-specific variant 4 (ETV4) in sorafenib and cisplatin resistance in hepatocellular carcinoma (HCC). Methods HCC cell lines SMMC-7721 and HCC-LM3 were transfected with an ETV4- overexpressing plasmid or small interfering RNAs (siRNAs) targeting ETV4. The cells with ETV4 overexpression or ETV4 interference were treated with DMSO, sorafenib (5 μmol/L) or cisplatin (5 μmol/L) for 48 h, and the total protein and total RNA were collected. Western blotting, flow cytometry, EdU proliferation assay were used to analyze the apoptosis and proliferation of the cells. We also obtained clinical specimens of HCC tissues and paired adjacent tissues from 11 patients for detecting ETV4 mRNA expression levels using real-time fluorescence quantitative PCR (q-PCR). The effect of ETV4 interference on the mRNA expression levels of immediate early response gene 3 (IER3) was examined in HCC cells that were treated with DMSO, sorafenib or cisplatin for 48 h. Results The expression of ETV4 mRNA was significantly higher in HCC tissues than in the paired adjacent tissues. Overexpression of ETV4 in the HCC cell lines obviously inhibited cell apoptosis induced by sorafenib or cisplatin. Conversely, ETV4 interference significantly enhanced the apoptosis and inhibited the proliferation of the HCC cells following treatments with sorafenib or cisplatin. In addition, ETV4 regulated the mRNA expression levels of IER3 in the cells treatmed with sorafenib and cisplatin. Conclusion ETV4 promotes resistance of HCC cells to sorafenib or cisplatin in vitro.