lncBRM对乳腺癌细胞生长的影响及作用机制  

Effect of lnc BRM on the growth of breast cancer cells and its mechanism

作　　者：杨净渝[1] 邱爽 陈昕[1] YANG Jingyu;QIU Shuang;CHEN Xin(Department of Breast and Thyroid Surgery,the First People's Hospital of Yunnan Province,Kunming 650032,China)

机构地区：[1]云南省第一人民医院乳腺甲状腺外科,昆明650032

出　　处：《中国癌症防治杂志》2020年第2期175-180,共6页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：云南省科技计划项目[2017FE467(-153)]。

摘　　要：目的探讨Brahma相关长链非编码RNA(long non-coding RNA for association with Brahma,lncBRM)对人乳腺癌细胞系(MCF-7、MDA-MB-453)生长、迁移和侵袭的影响及可能的作用机制。方法采用RT-PCR实验检测lncBRM在不同乳腺癌细胞(MCF-7、ZR-75-30、BT474、MDA-MB-231和MDA-MB-453)和正常人乳腺上皮细胞株MCF 10A中的表达;利用siRNA在MCF-7、MDA-MB-453细胞中敲低lncBRM,分别转染si-lncBRM质粒(si-lncBRM组)和空载质粒(si-Ctrl组),采用CCK-8实验检测细胞增殖情况,流式细胞术检测细胞凋亡情况,Transwell小室实验检测细胞侵袭和迁移能力,采用Western blot检测迁移相关蛋白E-cadherin和N-cadherin的表达水平,并采用miRDB数据库预测lncBRM靶点。结果 lncBRM在不同乳腺癌细胞中的表达水平均高于正常人乳腺上皮细胞株MCF 10A(P<0.01)。si-Ctrl组比较,敲低lncBRM可抑制MCF-7、MDA-MB-453细胞增殖能力(P<0.05),提高细胞凋亡率(P<0.01),细胞侵袭和迁移细胞数目较少(均P<0.01);同时E-cadherin表达明显上调,而N-cadherin表达下调(均P<0.01)。miRDB数据库预测发现lncBRM和68个miRNAs存在结合位点,敲低lncBRM导致评分最高的前5个miRNA中的4个(miR-4646-5p、miR-204-3p、miR-204-5p和miR-6832-3p)表达上调。结论 lncBRM可能通过调控miRNAs的表达而抑制乳腺癌细胞的增殖、侵袭和迁移并诱导其凋亡。Objective To investigate the effect of long non-coding RNA for association with Brahma(lncBRM)on the growth,migration,and invasion of human breast cancer cell lines(MCF-7,MDA-MB-453) and its possible mechanism. Methods The expression of lncBRM in different breast cancer cells(MCF-7,ZR-75-30,BT474,MDA-MB-231,and MDA-MB-453) and normal human breast epithelial cell line MCF 10 A was detected by RT-PCR. siRNA was used to knock down the expression of lncBRM in MCF-7,MDAMB-453 cells,and transfect si-lncBRM plasmid(si-lncBRM group)and blank plasmid(si-Ctrl group),respectively. CCK-8 assay was used to detect cell proliferation,and flow cytometry was used to detect cell apoptosis. The alteration of cell migration and invasion was detected by Transwell test. In addition,the changes in the expression of E-cadherin and N-cadherin were detected by Western blot.Target prediction of lncBRM was performed by miRDB database. Results The expression level of lncBRM in all breast cancer cells was significantly higher than that in normal human breast epithelial cell line MCF 10 A(P<0.01). Compared with the si-Ctrl group,lncBRM knockdown can inhibit the proliferation of MCF-7 and MDA-MB-453 cells(P<0.05),and increase the apoptosis rate(P<0.01),the numbers of cell invasion and migration cells were small(all P<0.01). Meanwhile,the protein expression of E-cadherin increased,while N-cadherin decreased dramatically(all P<0.01). The miRDB database predicted that lncBRM could target 68 miRNAs,and lncBRM knockdown could indeed increase the expressions of 4 miRNAs(miR-4646-5 p,miR-204-3 p,miR-204-5 p,and miR-6832-3 p)of the top 5 miRNAs. Conclusion lncBRM may inhibit the proliferation,invasion,and migration of breast cancer cells and induce their apoptosis by regulating the miRNAs.

关 键 词：乳腺癌 lncBRM 增殖 侵袭 迁移 MIRNA 

分 类 号：R737.9[医药卫生—肿瘤]