miR-363对顺铂耐药乳腺癌细胞的作用及机制  

Effect and Mechanism of mi R-363 on Cisplatin-treated Breast Cancer Cells

作　　者：吕晓皑[1] 王蓓[1] 陈建彬[1] 叶荆[1] 

机构地区：[1]浙江中医药大学附属第一医院乳腺病中心,杭州310006

出　　处：《中国现代应用药学》2015年第9期1041-1046,共6页Chinese Journal of Modern Applied Pharmacy

基　　金：浙江省中医药科技计划项目(2010KYA149)

摘　　要：目的研究miR-363对顺铂耐药乳腺癌细胞的作用及机制。方法采集以顺铂为主要化疗药物的中晚期乳腺癌患者血清,用定量PCR法检测化疗前和化疗后血清标本miR-363的表达水平。构建顺铂抵抗MCF-7细胞系(MCF-7-R),MTT法检测不同浓度顺铂对MCF-7及MCF-7-R细胞活力的影响。在MCF-7-R细胞中转染miR-363,MTT法检测miR-363是否能提高顺铂对MCF-7-R的杀伤活性。利用生物信息学、定量PCR及Western blot方法验证miR-363是否调节MCF-7-R细胞中Mcl-1的表达。构建Mcl-1真核表达载体,MTT法检测Mcl-1表达载体转染对miR-363联合顺铂治疗MCF-7-R疗效的影响。结果中晚期乳腺癌患者顺铂治疗后血清miR-363水平相较化疗前显著下降。MTT结果表明相同浓度顺铂对MCF-7-R的杀伤活性显著低于MCF-7细胞,且miR-363转染能显著提高顺铂对MCF-7-R的杀伤活性。生物信息学、定量PCR及Western blot结果表明miR-363转染可显著降低MCF-7-R细胞中Mcl-1的表达。miR-363联合顺铂在Mcl-1表达载体转染后对MCF-7-R细胞的杀伤活性显著低于未转染Mcl-1表达载体的miR-363联合顺铂组。结论 Mi R-363能增强耐顺铂乳腺癌细胞对顺铂杀伤作用的敏感性。OBJECTIVE To investigate the role and mechanism of miR-363 on cisplatin-resistant breast cancer cells. METHODS Collected the serum from the advanced breast cancer patients who were treated with cisplatin-based chemotherapy. Then the expression of miR-363 in breast cancer patients＇ serum was detected before or after chemotherapy using q PCR analysis. Constructed the cisplatin-resistant MCF-7 cell line, the cytotoxicity of cisplatin to MCF-7 cell line and MCF-7-R cell line was measured by MTT assay. Transfected the MCF-7-R cells with miR-363 to determine whether the transfection of miR-363 enhanced cytotoxicity of cisplatin to MCF-7-R cells. Confirmed whether the expression of McL^-1 was regulated by miR-363 using bioinformatics, q PCR and Western blot. Constructed the McL^-1 expression vector, and detected the cytotoxicity of cisplatin combing with the cotransfection of miR-363 and McL^-1 expression vector by MTT assay. RESULTS Mi R-363 levels were significantly decreased in advanced breast cancer patients treated with cisplatin-based chemotherapy. Mi R-363 levels were also lower in MCF-7-R cells than in MCF-7 cells. Exogenous miR-363 significantly overcame cisplatin resistance in MCF-7-R cells, whereas miR-363 knockdown increased the cell viability during cisplatin treatment. It was demonstrated that miR-363 directly targeted to McL^-1, and the downregulation of miR-363 resulted in upregulation of McL^-1. miR-363 decreased cisplatin resistance of MCF-7-R cells, partly by targeting McL^-1. CONCLUSION Downregulation of miR-363 increases drug resistance in cisplatin-treated MCF-7 by downregulating McL^-1.

关 键 词：MI R-363 MCL-1 MCF-7-R 顺铂 

分 类 号：R966[医药卫生—药理学]