lncRNA miR143HG抑制乳腺癌细胞增殖机制的初步研究  

Preliminary study on the inhibitory mechanism of lncRNA miR143HG on the proliferation of breast cancer cells

作　　者：徐沭芬 李娟[1] 陆彬彬[1] 皇甫超男 李杰 王科明[1] XU Shufen;LI Juan;LU Binbin;HUANGFU Chaonan;LI Jie;WANG Keming(Department of Oncology,the Second Affiliated Hospital of Nanjing Medical University,Nanjing 210011,China)

机构地区：[1]南京医科大学第二附属医院肿瘤科,南京210011

出　　处：《临床肿瘤学杂志》2020年第6期487-493,共7页Chinese Clinical Oncology

基　　金：南京市医学科技发展一般性课题资助项目(YKK16222);江苏省青年医学重点人才资助项目(QNRC2016662)。

摘　　要：目的研究长链非编码RNA miR143HG对乳腺癌细胞增殖、迁移、侵袭的影响及可能分子机制。方法采用实时荧光定量PCR(q PCR)检测乳腺癌细胞株MCF-7、MDA-MB-231和人正常乳腺上皮细胞MCF-10A的miR143HG表达量。过表达质粒载体(pc DNA-miR143HG,实验组)、空质粒载体(Empty vector,阴性对照组)分别转染MDA-MB-231和MCF-7细胞株,采用CCK-8、克隆形成实验检测细胞增殖活性,流式细胞术检测细胞凋亡和周期情况,Transwell实验检测细胞迁移、侵袭能力。裸鼠皮下移植瘤实验验证miR143HG在体内对乳腺癌细胞生长的影响。Western blotting检测各组p15、p16、p21、p27、p51蛋白表达情况。结果与MCF-10A细胞比较,MCF-7和MDA-MB-231细胞中miR143HG表达量降低(P<0. 05)。CCK-8法结果显示实验组细胞增殖活性明显低于对照组(P<0. 05);克隆形成实验显示,实验组细胞集落形成数量低于对照组(P<0. 05);流式细胞术结果显示实验组细胞凋亡率高于对照组(P<0. 05),实验组G0/G1期细胞比例高于对照组(P<0. 05);Transwell结果显示实验组迁移和侵袭细胞数低于对照组(P<0. 05)。裸鼠皮下移植瘤实验显示,实验组移植瘤体积小于对照组,实验组移植瘤质量低于对照组[(0. 27±0. 11) g vs.(0. 58±0. 20) g],差异具有统计学意义(P<0. 05)。Western blotting结果显示,实验组细胞中p21蛋白表达水平高于对照组(P<0. 05)。结论 lncRNA miR143HG可能通过上调p21的表达来发挥抑制乳腺癌细胞增殖、迁移侵袭、促进细胞周期停滞和凋亡的作用。Objective To study the effect of long non-coding RNA miR143HG on proliferation,migration and invasion of breast cancer cells and its possible molecular mechanism.Methods The expression of miR143HG in breast cancer cell lines MCF-7,MDA-MB-231 and human normal breast epithelial cells MCF-10A were detected by real-time fluorescent quantitative PCR(qPCR).MDA-MB-231 and MCF-7 cell lines were transfected with overexpressed plasmid vector(pcDNA-miR143HG,experimental group)and Empty vector(negative control group),respectively.Cell viability was detected by CCK-8 and clonal formation experiments,apoptosis and cell cycle were detected by flow cytometry,and cell migration and invasion were detected by Transwell experiment.The effect of miR143HG on the growth of breast cancer cells in vivo was confirmed by subcutaneous tumor transplantation in nude mice.Protein expressions of p15,p16,p21,p27 and p51 in each group were detected by Western blotting.Results Compared with MCF-10A,expression of miR143HG was decreased in MCF-7 and MDA-MB-231 cells(P<0.05).The results of CCK-8 method showed that the cell vitality of the experimental group was significantly lower than that of the negative control group(P<0.05).The number of colony formation in the experimental group was lower than that in the negative control group(P<0.05).The results of flow cytometry showed that the apoptosis rate of the experimental group was higher than that of the negative control group(P<0.05).The proportion of G 0/G 1 phase cells in the experimental group was higher than that in the negative control group(P<0.05).Transwell results showed that the number of cell migration and invasion in the experimental group was lower than that in the negative control group(P<0.05).Subcutaneous tumor transplantation in nude mice showed that the tumor volume of the experimental group was smaller than that of the negative control group,and the tumor weight of the experimental group was lower than that of the negative control group[(0.27±0.11)g vs.(0.58±0.20)g],and the differ

关 键 词：乳腺癌 miR143HG 增殖 细胞周期 迁移 侵袭 P21 

分 类 号：R737.9[医药卫生—肿瘤]