miR-363通过靶向于MCL-1增强顺铂对乳腺癌细胞的杀伤活性  

Inducing the viability inhibition of cisplatin in breast cancer cells by miR-363 targeting MCL-1

作　　者：徐峰[1] 章蔚[1] 田景琦[1] 

机构地区：[1]绍兴市人民医院乳腺甲状腺外科,浙江绍兴312000

出　　处：《中国卫生检验杂志》2015年第19期3236-3239,共4页Chinese Journal of Health Laboratory Technology

摘　　要：目的探究microRNA-363(miR-363)是否能增强顺铂对乳腺癌细胞的杀伤活性及机制。方法利用生物信息学、定量PCR及Western blot法,验证miR-363能否在体外调节乳腺癌细胞系MDA-MB-231 MCL-1的表达水平;MTT法检测miR-363联合顺铂对乳腺癌细胞系MDA-MB-231的杀伤活性;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联合顺铂治疗乳腺癌疗效的影响。结果乳腺癌细胞系MDA-MB-231的miR-363表达水平显著低于正常乳腺细胞系MCF-10A,差异有统计学意义(P<0.05)。miR-363转染后,乳腺癌细胞系MDA-MB-231 MCL-1的mRNA表达水平及蛋白表达水平相比于NCO转染组均下降,差异有统计学意义(P<0.05)。MTT结果表明,miR-363联合顺铂组对MDA-MB-231细胞的杀伤活性显著高于miR-363组和顺铂组,差异有统计学意义(P<0.05)。miR-363联合顺铂在MCL-1表达载体转染后对MDA-MB-231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,差异有统计学意义(P<0.05)。结论 miR-363通过靶向与MCL-1增强顺铂对乳腺癌细胞的杀伤活性。Objective To study if miR- 363 can induce the viability inhibition of cisplatin in breast cancer cell and its machanism. Methods Bioinformatics,quantitative PCR and Western blot were used to evaluate whether miR- 363 can regulate the expression level of MDA- MB- 231 MCL- 1 in breast cancer cell in vitro. MTT assay was performed to measure the viability of MDA- MB- 231 cell treated with miR- 363 plus cisplatin. MCL- 1 expression vector was constructed,and then the viability of MDA- MB- 231 cell was detected by MTT assay to evaluate the effect of MCL- 1 expression vector,toward miR- 363 plus cisplatin,on breast cancer. Results The expression level of miR- 363 was lower in MDA- MB- 231 cell line than that in the normal breast cell line MCF- 10 A,and the difference had statistical significance（ P〈0. 05）. The expression of MCL- 1 at both mRNA level and protein level was downregulated after miR- 363 transfection in MDA- MB- 231 cells,compared with that in the cells transfected with NCO,and the difference was statistically significant（ P〈0. 05）. The results of MTT assay showed that the viability of MDA- MB- 231 cell in miR- 363 plus cisplatin group was significantly lower than that in miR-363 group and cisplatin group,and the difference was statistically significant（ P〈0. 05）. Furthermore,the viability inhibition of MDA- MB- 231 cells treated with miR- 363 plus cisplatin significantly decreased after the transfection of MCL- 1 expression vector,which was lower than that in the one without transfection,and the difference was statistically significant（ P〈0. 05）. Conclusion miR- 363 targeting MCL- 1 can induce the viability inhibition of cisplatin in breast cancer cell.

关 键 词：microRNA-363 MCL-1 乳腺癌 顺铂 

分 类 号：R737.9[医药卫生—肿瘤]