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miR-130a-5p靶向TPD52对人乳腺癌细胞增殖和凋亡的影响OA

Effect of miR-130a-5p targeting TPD52 on proliferation and apoptosis of human breast cancer cells

中文摘要英文摘要

目的 探讨miR-130a-5p靶向调控肿瘤蛋白D52(TPD52)对人乳腺癌细胞增殖和凋亡的影响.方法 将人乳腺癌MCF-7细胞分为对照组、NC mimic组(转染模拟物阴性对照)、miR-130a-5p mimic组(转染miR-130a-5p模拟物)、miR-130a-5p mimic+pcDNA-NC组(转染miR-130a-5p模拟物和pcDNA阴性对照)、miR-130a-5p mimic+pcDNA-TPD52组(转染miR-130a-5p模拟物和pcDNA-TPD52).采用生物信息学软件及双荧光素酶报告实验分析并验证miR-130a-5p对TPD52的靶向关系,荧光定量PCR法检测各组细胞miR-130a-5p和TPD52 mRNA表达情况,细胞计数试剂盒-8(CCK-8)检测各组细胞增殖能力,流式细胞术和4',6-二脒基-2-苯基吲哚(DAPI)核染法检测细胞凋亡率和核凋亡情况,Western blot测定增殖相关蛋白、凋亡相关蛋白表达情况.结果 miR-130a-5p和TPD52的3'-UTR存在连续的结合位点,miR-130a-5p可靶向调控TPD52的表达.与NC mimic组相比,miR-130a-5p mimic组细胞中miR-130a-5p的相对表达水平明显升高,TPD52 mRNA相对表达水平明显降低(P<0.05);与miR-130a-5p mimic+pcDNA-NC组比较,miR-130a-5p mimic+pcDNA-TPD52组细胞中TPD52 mRNA相对表达水平明显升高(P<0.05).与NC mimic组细胞比较,miR-130a-5p mimic组细胞存活率明显降低,细胞凋亡率明显升高(P<0.05);与 miR-130a-5p mimic+pcDNA-NC组细胞比较,miR-130a-5p mimic+pcDNA-TPD52组细胞存活率明显升高,细胞凋亡率明显降低(P<0.05).与NC mimic组比较,miR-130a-5p mimic组细胞中Bax、caspase 3蛋白表达水平升高,PCNA蛋白表达水平降低(P<0.05);与 miR-130a-5p mimic+pcDNA-NC组比较,miR-130a-5p mimic+pcDNA-TPD52组细胞中Bax、caspase 3蛋白表达水平降低,而PCNA蛋白表达水平升高(P<0.05).结论 miR-130a-5p通过靶向调控TPD52,进而抑制MCF-7细胞增殖,诱导MCF-7细胞凋亡.

Objective To investigate the effect of miR-130a-5p targeting tumor protein D52(TPD52)on proliferation and apoptosis of human breast cancer cells.Methods Human breast cancer MCF-7 cells were randomly divided into the control group,the NC mimic group(transfected with mimic negative control),the miR-130a-5p mimic group(transfected with miR-130a-5p mimic),the miR-130a-5p mimic+pcDNA-NC group(co-transfected with miR-130a-5p mimic and pcDNA negative control),and the miR-130a-5p mimic+pcDNA-TPD52 group(co-transfected with miR-130a-5p mimic and pcDNA-TPD52).Bioinformatics software and dual-luciferase reporter assay were used to analyze and validate the targeting relationship of miR-130a-5p to TPD52.Quantitative real-time PCR was used to detect the mRNA expression of miR-130a-5p and TPD52 mR-NA in each group.Cell counting kit-8(CCK-8)assay was used to assess cell proliferation.Flow cytometry and 4',6-diamidino-2-phenylindole(DAPI)nuclear staining were used to detect apoptosis rate of cells and nuclear apoptosis.Western blot was used to detect the expression of proliferation-related and apoptosis-related pro-teins.Results There were consecutive binding sites between miR-130a-5p and the 3'-UTR of TPD52,and miR-130a-5p could target and regulate TPD52 expression.Compared with the NC mimic group,the relative expression level of miR-130a-5p in the miR-130a-5p mimic group was significantly increased,while the relative expression level of TPD52 mRNA was significantly decreased(P<0.05).Compared with the miR-130a-5p mimic+pcDNA-NC group,the relative expression level of TPD52 mRNA in the miR-130a-5p mimic+pcDNA-TPD52 group was significantly increased(P<0.05).Compared with the NC mimic group,the cell viability in the miR-130a-5p mimic group was significantly decreased(P<0.05),while the apoptosis rate was significant-ly increased(P<0.05).Compared with the miR-130a-5p mimic+pcDNA-NC group,the cell viability in the miR-130a-5p mimic+pcDNA-TPD52 group was significantly increased(P<0.05),while the apoptosis rate was significantly decreased(P<0.05).Compared with the NC mimic group,the expression levels of Bax and caspase 3 proteins in the miR-130a-5p mimic group were significantly increased(P<0.05),while the expres-sion level of PCNA protein was significantly decreased(P<0.05).Compared with the miR-130a-5p mimic+pcDNA-NC group,the expression levels of Bax and caspase 3 proteins in the miR-130a-5p mimic+pcDNA-TPD52 group were significantly decreased(P<0.05),while the expression level of PCNA protein was signifi-cantly increased(P<0.05).Conclusion miR-130a-5p inhibits MCF-7 cell proliferation and induces apoptosis by targeting TPD52.

管福强;余佳勉;劳俊杰;余道军

浙江中医药大学研究生院,杭州 310053||杭州市职业病防治院检验科,杭州 310011杭州市职业病防治院检验科,杭州 310011西湖大学医学院附属杭州市第一人民医院检验科,杭州 310006西湖大学医学院附属杭州市第一人民医院检验科,杭州 310006

医药卫生

miR-130a-5p肿瘤蛋白D52乳腺癌增殖凋亡

miR-130a-5ptumor protein D52breast cancerproliferationapoptosis

《重庆医学》 2026 (8)

1736-1741,6

10.3969/j.issn.1671-8348.2026.08.009

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