下调HMGA1对乳腺癌MCF7细胞增殖和侵袭能力的影响及机制  

Effects of down-regulation of HMGA1 on proliferation and invasion of breast cancer MCF7 cells

作　　者：张洋[1] 董理[2] 孙源博 李文媛[1] 王莹[1] 孙平[1] 于伟光[2] 

机构地区：[1]牡丹江医学院,黑龙江牡丹江157011 [2]牡丹江医学院红旗医院

出　　处：《山东医药》2017年第38期15-18,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81371362);黑龙江省自然科学基金项目(H201377)

摘　　要：目的观察下调高迁移率族蛋白A1(HMGA1)对乳腺癌MCF7细胞增殖和侵袭能力的影响,并探讨其机制。方法选择人乳腺癌MCF7细胞,将其分为siRNA对照组、siRNA-HMGA1组、miR对照组和miR-24-3p组,分别转染siRNA对照慢病毒、siRNA-HMGA1慢病毒、miR对照慢病毒和miR-24-3p慢病毒。采用Western boltting法检测未转染MCF7、正常乳腺上皮细胞株(HMEC h TERT)及siRNA对照组、siRNA-HMGA1组细胞HMGA1蛋白,实时荧光定量PCR法检测miR对照组和miR-24-3p组细胞HMGA1、miR-142-3p mRNA,MTT法和Transwell法检测4组细胞增殖活力和侵袭能力,荧光素酶报告基因分析验证HMGA1与miR-142-3p靶向关系。结果 MCF7、HMEC h TERT细胞中HMGA1蛋白相对表达量分别为0.574±0.075、0.234±0.099,两者相比P<0.01。siRNA-HMGA1组、siRNA对照组HMGA1蛋白相对表达量分别为0.141±0.051、0.651±0.136,两组相比P<0.01。miR-142-3p组、miR对照组HMGA1 mRNA相对表达量分别为0.135±0.046、0.604±0.156,miR-142-3 mRNA相对表达量分别为1.128±0.174、0.263±0.116,两者相比P均<0.01。MCF7细胞中HMGA1 mRNA、miR-142-3p mRNA表达呈负相关(r=-0.259,P=0.021)。siRNA-HMGA1组、siRNA对照组细胞增殖活力分别为0.535±0.066、1.160±0.125,穿膜细胞数分别为(32.05±9.33)、(71.68±14.39)个,两者相比P均<0.01。miR-142-3p组、miR对照组细胞增殖活力分别为0.362±0.121、1.083±0.139,穿膜细胞数分别为(21.52±6.64)、(46.74±7.82)个,两组相比P均<0.01。miR-142-3p组、miR对照组野生型HMGA1 3'-UTR荧光素酶活性分别为0.668±0.074、1.000±0.000,两组相比P<0.01。结论下调HMGA1乳腺癌MCF7细胞增殖和侵袭能力增强,其机制可能与HMGA1可靶向反向调控miR-142-3p有关。Objective To investigate the effects of the down-regulation of high mobility group protein A1 （HMGA1）on the proliferation and invasion of breast cancer MCF7 cells and its mechanism.Methods The human breast cancer cell line （MCF7） was divided into four groups： siRNA control group,siRNA-HMGA1 group,miR control group,and miR-24-3p group; MCF7 cells in the above groups were transfected with siRNA control lentivirus,siRNA-HMGA1 lentivirus,miR control lentivirus and miR-24-3p lentivirus,respectively.The protein expression of HMGA1 was detected in MCF7 cells,normal breast epithelial cell line （HMEC h TERT）,siRNA control group,and siRNA-HMGA1 group by using Western blotting; the HMGA1 and miR-142-3p mRNA expression and its correlation was detected in the miR control group and miR-24-3p group by real-time quantitative PCR; the cell proliferation and invasion were detected in the four groups by MTT and Transwell assay; Luciferase reporter experiment was used to verify the target relationship between HMGA1 and miR-142-3p.Results HMGA1 protein expression in MCF7 cells was 0.574 ± 0.075,which was significantly higher than that of HMEC h TERT cells （0.234 ± 0.099） （P〈0.05）,and the expression of HMGA1 in siRNA-HMGA1 group （0.141 ±0.051） was lower than that of siRNA group （0.651 ± 0.136） （P〈0.05）; HMGA1 mRNA expression in the miR-142-3p group was lower than that of the miR control group （0.135 ± 0.046 vs 0.604 ± 0.156）,and the miR-142-3 mRNA expression was higher than that of the miR control group （1.128 ± 0.174 vs 0.263 ± 0.116） （all P〈0.01）; the expression of HMGA1 mRNA and miR-142-3p mRNA was negatively correlated （r = 0.259,P〈0.05）; the cell proliferation in the siRNA-HMGA1 group was lower than that of the control group （0.535 ± 0.066 vs 1.160 ± 0.125）,transmembrane cell number was also lower than that of the siRNA group （32.05 ± 9.33 vs 71.68 ± 14.39） （P〈0.05）; the cell proliferation ability of miR-142-3p group was lower than that of the miR

关 键 词：高迁移率族蛋白A1 微小RNA 142-3p 乳腺肿瘤 细胞增殖 细胞侵袭 

分 类 号：R735[医药卫生—肿瘤]