lncRNA VIM-AS5在人乳腺癌细胞系中的表达及对乳腺癌细胞增殖和迁移的影响  

lncRNA VIM-AS5 expression and its effect on proliferation and migration of human breast cancer cell lines

作　　者：陆凯 陆建菊 郭文利 黄建棋 李志华 LU Kai;LU Jianju;GUO Wenli;HUANG Jianqi;LI Zhihua(Department of Breast Diseases,the First Hospital of Jiaxing,Jiaxing 314001;Department of Breast Oncology,the Third Hospital of Nanchang,Nanchang 330008,China)

机构地区：[1]嘉兴市第一医院乳腺病科,浙江嘉兴314001 [2]南昌市第三医院乳腺肿瘤科,江西南昌330008

出　　处：《基础医学与临床》2024年第4期447-453,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81860546);嘉兴市科技计划项目(2020AD30066)。

摘　　要：目的 探讨长链非编码RNA (lncRNA)VIM-AS5在人乳腺癌组织中表达的临床意义及其对乳腺癌细胞增殖和迁移的作用机制。方法 利用Lnc2Cancer 3.0数据库分析VIM-AS5在乳腺癌组织中的表达及其与乳腺癌患者临床分期、生存期的相关性。RT-qPCR检测乳腺癌BT-549、MDA-MB-435、MDA-MB-231、CAL-51细胞中VIM-AS5的表达量。通过脂质体分别将VIM-AS5过表达质粒和阴性对照质粒转染至CAL-51细胞,依次记为VIM-AS5组和NC组。采用集落形成实验、划痕愈合法分别检测CAL-51细胞的增殖活力和迁移率。双荧光素酶报告基因实验验证VIM-AS5与miR-500a的靶向关系。RT-qPCR检测CAL-51细胞中miR-500a表达。Western blot检测CAL-51细胞中JAK/STAT3分子通路蛋白表达。结果 乳腺癌组织中VIM-AS5表达显著低于癌旁组织(P<0.01)。VIM-AS5表达量与乳腺癌患者的临床分期呈负相关(P<0.01)。VIM-AS5低表达乳腺癌患者的生存期明显短于VIM-AS5高表达乳腺癌患者(P<0.01)。与乳腺上皮MCF-10A细胞相比,乳腺癌细胞中VIM-AS5表达显著降低(P<0.01)。VIM-AS5组集落形成数显著低于NC组(P<0.01)。VIM-AS5组细胞迁移率显著低于NC组(P<0.01)。双荧光素酶报告基因实验证实miR-500a是VIM-AS5的靶基因(P<0.01)。VIM-AS5能够负调控miR-500a的表达(P<0.01)。与NC组比较,VIM-AS5组CAL-51细胞中JAK/STAT3分子通路蛋白JAK、p-STAT3、c-Myc、Bcl-2、CDK3表达均明显下降。结论 乳腺癌细胞中VIM-AS5呈低表达,上调VIM-AS5可能通过靶向miR-500a/JAK/STAT3分子通路抑制乳腺癌CAL-51细胞的增殖及迁移。Objective To explore the clinical significance of long non-coding RNA(lncRNA)VIM-AS5 expression in human breast cancer tissues and its regulatory mechanism involved in cancer cell proliferation and migration.Methods The Lnc2Cancer 3.0 database was used to analyze the expression of VIM-AS5 in breast cancer tissues and its correlation with the clinical stage and survival time of breast cancer patients.RT-qPCR was used to detect the expression of VIM-AS5 in breast cancer cell lines BT-549,MDA-MB-435,MDA-MB-231 and CAL-51.Plasmid with VIM-AS5 overexpression and negative control were all transfected into CAL-51 cells through liposome recorded as VIM-AS5 group and NC group,respectively.The proliferation and migration of CAL-51 cells were detected by colony formation assay and scratch healing method,respectively.Dual-luciferase reporter gene experiment verified the targeting relationship between VIM-AS5 and miR-500a.RT-qPCR was used to detect the expression of miR-500a in CAL-51 cells.Western blot was used to detect the expression of JAK/STAT3 pathway in CAL-51 cells.Results The expression of VIM-AS5 in breast cancer tissues was significantly lower than that in adjacent tissues(P<0.01).VIM-AS5 expression was negatively correlated with the clinical stage of breast cancer patients(P<0.01).The survival time of breast cancer patients with low VIM-AS5 expression was significantly shorter than that of breast cancer patients with high VIM-AS5 expression(P<0.01).Compared with mammary epithelial cell line MCF-10A cells,VIM-AS5 expression was significantly reduced in breast cancer cells(P<0.01).The counting number of colony formed in the VIM-AS5 group was significantly lower than that in the NC group(P<0.01).The cell migration rate in the VIM-AS5 group was significantly lower than that in the NC group(P<0.01).Dual-luciferase reporter gene experiment confirmed that miR-500a was the target gene of VIM-AS5(P<0.01).VIM-AS5 can negatively regulate the expression of miR-500a(P<0.01).Compared with the NC group,the expression of JAK/STAT

关 键 词：lncRNA VIM-AS5 乳腺癌 miR-500a 细胞增殖 细胞迁移 

分 类 号：R737.9[医药卫生—肿瘤]