LncRNA NEAT1通过靶向Hsa-miR-206调控乳腺癌细胞增殖、侵袭和迁移的实验研究  

LncRNA NEAT1 Regulated Proliferation,Invasion and Migration of Breast Cancer Cells by Targeting Hsa-miR-206

作　　者：魏涛[1] 冯连杰[1] 付小娜 赵世杰[1] 刘伟鹏 梁豪 WEI Tao;FENG Lian-jie;FU Xiao-na(Anyang Cancer Hospital,Anyang Henan 455000,China)

机构地区：[1]安阳市肿瘤医院放疗三科,河南安阳455000

出　　处：《湖北科技学院学报（医学版）》2021年第6期496-500,508,F0003,共7页Journal of Hubei University of Science and Technology(Medical Sciences)

基　　金：安阳市科技发展计划(119)。

摘　　要：目的探讨长链非编码RNA(lncRNA)NEAT1通过靶向miR-206对乳腺癌细胞增殖、侵袭和迁移的影响。方法培养人乳腺癌细胞和乳腺上皮细胞,RT-qPCR检测细胞中NEAT1和miR-206的表达水平,双荧光素酶报告基因实验验证NEAT1与miR-206的靶向结合关系,MTT实验和Transwell实验分别检测敲减NEAT1和抑制miR-206对乳腺癌细胞增殖、迁移和侵袭的影响,Western blot检测敲减NEAT1和抑制miR-206对乳腺癌细胞中基质金属蛋白酶2(MMP-2)和MMP-9蛋白表达的影响。结果与人乳腺上皮细胞相比,乳腺癌细胞中NEAT1表达上调(P<0.05),miR-206表达下调(P<0.05);RT-qPCR结果表明,敲减NEAT1可促进乳腺癌细胞中miR-206表达,而过表达NEAT1可抑制乳腺癌细胞中miR-206表达;双荧光素酶报告基因实验证实NEAT1靶向负调控miR-206的表达;敲减NEAT1明显抑制乳腺癌细胞增殖、侵袭和迁移能力及细胞中MMP-2和MMP-9蛋白表达,而敲减NEAT1的同时抑制miR-206则可逆转敲减NEAT1对细胞增殖、侵袭和迁移的抑制作用,逆转敲减NEAT1对细胞中MMP-2和MMP-9蛋白表达的抑制作用。结论敲减NEAT1可能通过靶向抑制miR-206表达,发挥抑制乳腺癌细胞增殖、侵袭和迁移的作用。Objective To investigate the effects of long non-coding RNA(lncRNA)NEAT1 on proliferation,invasion and migration of breast cancer cells by targeting miR-206.Methods Cultured human breast cancer cells and mammary epithelial cells,and the expression levels of NEAT1 and miR-206 in cells were detected by RT-qPCR,the targeted binding relationship between NEAT1 and miR-206 was verified by Dual luciferase reporter gene assay,the effect of knockdown of NEAT1 and inhibition of miR-206 on proliferation,migration and invasion of breast cancer cells was detected by MTT assay and Transwell asssy.Western blot was used to detect knockdown of NEAT1 and inhibition of miR-206 on the expression of matrix metalloproteinase-2(MMP-2)and MMP-9 protein in breast cancer cells.Results Compared with human mammary epithelial cells,the expression of NEAT1 was up-regulated in breast cancer cells(P<0.05),and the expression of miR-206 was down-regulated(P<0.05);RT-qPCR results showed that knockdown of NEAT1 promoted miR-206 expression in breast cancer cells,while overexpression of NEAT1 inhibited miR-206 expression in breast cancer cells;Dual luciferase reporter gene experiments confirmed that NEAT1 targets negative regulation the expression of miR-206;Knockdown of NEAT1 significantly inhibited the proliferation,invasion and migration of breast cancer cells and the expression of MMP-2 and MMP-9 proteins in cells,while knockdown of NEAT1 and inhibition of miR-206 reversed knockdown of NEAT1 on cell proliferation,invasion and migration.Inhibition,reverse knockdown of NEAT1 inhibits MMP-2 and MMP-9 protein expression in cells.Conclusion Knockdown of NEAT1 might inhibit the proliferation,invasion and migration of breast cancer cells by targeting inhibition of miR-206 expression.

关 键 词：乳腺癌 LncRNA NEAT1 Hsa-miR-206 增殖 侵袭 迁移 

分 类 号：R737.9[医药卫生—肿瘤]