miR-149-5p靶向DCLK1基因对乳腺癌细胞MCF-7/DDP顺铂耐药的影响  

Effect of miR-149-5p targeting DCLK1 gene on cisplatin resistance of breast cancer cells MCF-7/DDP and its mechanism

作　　者：朱会利[1] 王清云 ZHU Hui-li;WANG Qing-yun(General Surgery Department of Handan Hospital of Integrated Traditional Chinese and Western Medicine,Handan 056001,China;Laboratory Department,The First Affiliated Hospital of Xingtai Medical College,Xingtai 054001,China)

机构地区：[1]邯郸市中西医结合医院普外科,河北邯郸056001 [2]邢台医学高等专科学校第一附属医院检验科,河北邢台054001

出　　处：《生物技术》2023年第2期202-207,共6页Biotechnology

基　　金：邢台市重点研发计划项目(2020ZC198)。

摘　　要：[目的]探讨miR-149-5p靶向双皮质素样激酶1(DCLK1)基因对乳腺癌细胞MCF-7/DDP顺铂耐药的影响。[方法]构建DDP耐药乳腺癌MCF-7细胞,然后按细胞转染质粒的不同分入对照组(不转染质粒)、空载组(转染空载质粒3.1)、miRNA-149-5p组(转染miRNA-149-5p-AH质粒)。采用CCK-8法检测细胞存活率,划痕实验检测细胞迁移力,流式细胞术检测细胞凋亡率,实时荧光定量PCR检测细胞miRNA-149-5p和DCLK1 mRNA水平,Western Bloting检测细胞DCLK1蛋白表达水平。[结果] miRNA-149-5p组乳腺癌MCF-7/DDP细胞miRNA-149-5p水平、细胞凋亡率显著高于对照组和空载组,DCLK1 mRNA和蛋白水平、细胞存活率和细胞迁移率显著低于对照组和空载组,差异均有统计学意义(P<0.05)。空载组和对照组miRNA-149-5p水平、细胞凋亡率、DCLK1 mRNA和蛋白表达、细胞存活率和细胞迁移率比较差异均无统计学意义(P>0.05)。[结论]过表达miRNA-149-5p可减弱乳腺癌MCF-7/DDP细胞的顺铂耐药性,使乳腺癌细胞增殖和迁移率降低,凋亡增加,其机制可能与靶向抑制DCLK1的表达有关。[Objective]To investigate the effect of miR-149-5p targeting doublecortin-like kinase 1(DCLK1) gene on MCF-7/DDP cisplatin resistance in breast cancer cells.[Method]DDP-resistant breast cancer MCF-7 cells were constructed,and then divide them into the control group(without transfection plasmid),the empty group(transfected with empty plasmid 3.1),and miRNA-149-5p group(transfected with miRNA-149-5p-AH plasmid) according to the different plasmids of the cells.CCK-8 method was used to detect cell viability,scratch test to detect cell migration,flow cytometry to detect cell apoptosis,real-time fluorescent quantitative PCR to detect miRNA-149-5p and DCLK1 mRNA levels,and Western bloting to detect DCLK1 protein expression level in cells of all groups.[Result]The level of miRNA-149-5p in breast cancer MCF-7/DDP cells in miRNA-149-5p group and apoptosis rate were significantly higher than the control and empty groups,DCLK1 mRNA and protein levels,cell viability and cell mobility were significantly lower than the control group and the no-load group,and the differences were statistically significant(P<0.05).The levels of miRNA-149-5p in the empty load group and control group,apoptosis rate,DCLK1 mRNA and protein expression,cell viability and cell mobility was not statistically significant(P<0.05).[Conclusion] Overexpression of miRNA-149-5p can attenuate the cisplatin resistance of breast cancer MCF-7/DDP cells,reduce the proliferation and migration rate of breast cancer cells,and increase apoptosis.The mechanism may be related to the targeted inhibition of DCLK1 expression.

关 键 词：miRNA-149-5p 双皮质素样激酶1 乳腺癌 顺铂耐药 

分 类 号：R737.9[医药卫生—肿瘤]