顺铂通过促进microRNA-214表达抑制肝癌细胞增殖  

Cisplatin suppresses human hepatocellular carcinoma cell growth via regulating microRNA-214-mediated β-catenin axis

作　　者：刘子文[1] 杜永星 由磊[1] 舒红[1] 赵玉沛[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院基本外科,北京100730

出　　处：《基础医学与临床》2014年第9期1199-1203,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(81272767)

摘　　要：目的研究顺铂对microRNA-214(miR-214)在肝癌细胞中表达的影响及其抑制肝癌细胞增殖的分子机制。方法用real-time PCR方法分析miR-214在肝细胞癌组织及细胞系中的表达;利用CCK-8细胞增殖分析法确定顺铂在肝癌细胞系Hep G2及Hep3b中的半数致死浓度(IC50),并用real-time PCR和Western blot方法检测经不同浓度顺铂处理的肝癌细胞Hep G2及Hep3b中miR-214及其靶基因β-catenin的表达变化;设计拯救实验研究顺铂、miR-214与肝癌细胞增殖的关系。结果 miR-214在肝细胞癌组织及细胞系中表达下调(P<0.01);顺铂处理的肝癌细胞系Hep G2、Hep3b内源性miR-214表达水平明显上调(P<0.01),而其靶基因β-catenin的表达水平明显下调(P<0.05)。结论顺铂通过调控miR-214介导的β-catenin表达发挥抑制肝癌细胞增殖的作用。Objective To investigate the impact of cisplatin on microRNA-214 （miR-214） expression in human hepatocellular carcinoma （HCC） cell lines and to elucidate the molecular mechanism of cisplatin for HCC chemotherapy. Methods Real-time PCR was conducted to determine the expression level of miR-214 in HCC tissue specimens and HCC cell lines. The IC50 value of cisplatin was determined both in HepG2 and Hep3b cell lines. The expression of miR-214 was evaluated in HepG2 and Hep3b cells with cisplatin treatment by using real-time PCR and we also performed Western blot analysis to detect the protein levels of β-catenin in HCC cells with the same treatment as described above. A rescue assay was conducted by using cisplatin treatment in combination with miR-214 inhibitor （Anti-214） to further investigate the correlation among cisplatin, miR-214, and cell growth. Results miR-21g expression depicted a significant downregulation in HCC tis- sues and cell lines （P 〈 0.01 ）. Cisplatin treatment led to an augment of miR-21g expression in HepG2 and Hep3b cells （ P 〈 0.01 ） and resulted in a decrease of β-catenin protein levels, which was verified as a direct target of miR-214 in HCC cells （ P 〈 0.05 ）. Conclusions Cisplatin represents its suppressive effect on HCCgrowth at least partially through regulation of miR-214-mediated β-catenin axis.

关 键 词：miR-214 肝细胞癌 化疗 

分 类 号：R735.7[医药卫生—肿瘤]