基础医学与临床 ›› 2019, Vol. 39 ›› Issue (10): 1467-1473.

• 研究论文 • 上一篇    下一篇

miR-488-3p下调BRCA2蛋白促进乳腺癌细胞系MCF7的侵袭与增殖

马秋晟1,刘峰2,方贵龙1   

  1. 1. 天津北辰医院
    2. 深圳市龙华区中心医院
  • 收稿日期:2018-09-21 修回日期:2019-01-23 出版日期:2019-10-05 发布日期:2019-09-25
  • 通讯作者: 方贵龙 E-mail:3623452139@qq.com

miR-488-3p promotes invasion and proliferation of breast cancer cell line MCF7 through inhibiting BRCA2 expression

  • Received:2018-09-21 Revised:2019-01-23 Online:2019-10-05 Published:2019-09-25

摘要: 目的 探讨miR-488-3p的表达与乳腺癌患者临床特征的相关性,及通过抑制易感基因(BRCA2)促进乳腺癌细胞侵袭、增殖、迁移及细胞周期在S期聚集的作用。方法 RT-qPCR检测乳腺癌组织、远端转移肿瘤组织和未转移肿瘤组织miR-488-3p的表达;检验BRCA2是否为miR-488-3p的潜在靶基因;RT-qPCR检测乳腺癌组织BRCA2 mRNA表达量;Western blot检测乳腺癌肿瘤组织BRCA2蛋白表达;用CCK8法检测吸光光度值;用流式细胞计量术检测细胞周期S期的细胞比值;用Tranwell小室法检测乳腺癌细胞系MCF7的侵袭、增殖、迁移及S期的聚集;构建乳腺癌异种移植裸鼠模型,检测裸鼠肿瘤体积及重量及miR-488-3p在miR-488-3p模拟物转染组裸鼠肿瘤组织中的表达量。结果 在乳腺癌细胞miR-488-3p表达显著增高(P<0.01);在远端转移肿瘤组织中明显高表达(P<0.01);miR-488-3p与淋巴结转移及TNM分期表达呈明显相关(P<0.05);miR-488-3p能抑制BRCA2的mRNA翻译及其蛋白表达。BRCA2对乳腺癌有抑制效果,可促进乳腺癌细胞MCF7的侵袭、增殖、迁移及在细胞周期S期聚集。上述作用可被BRCA2抑制;在裸鼠体内,miR-488-3p能促进MCF7细胞的增殖。结论 miR-488-3p抑制BRCA2,促进乳腺癌细胞增殖,有可能成为乳腺癌诊疗的新靶向。

关键词: 乳腺癌, miR-488-3p, BRCA2

Abstract: Objective To explore the correlation between the expression of miR-488-3p and the clinical features of breast cancer patients, and to promote breast cancer cell invasion, proliferation, migration and cell cycle aggregation in S phase by inhibiting BRCA2. Methods The expression of mir-488-3p in breast cancer tissue samples was detected by RT-qPCR. BRCA2 was tested as a potential target gene for miR-488-3p. The effect of RT-qPCR on mRNA expression of BRCA2 in breast cancer tissues; The protein expression level of BRCA2 in breast cancer tumor tissues was detected by Western blot. OD value of each transfection group was detected by CCK8 assay experiment. The cell ratio of each transfection group at S phase was determined by flow cytometry. Tranwell migration were used to detect the invasion, proliferation, migration and aggregation of MCF7 in breast cancer cell line. A xenograft model of breast cancer xenograft in nude mice was established to detect tumor volume and weight in nude mice. The expression of miR-488-3p in tumor tissues of nude mice transfected with miR-488-3p mimics was detected. Results miR-488-3p was highly expressed in breast cancer tumor tissues. miR -488-3p was also highly expressed in distal metastatic tumor tissues. miR-488-3p was significantly correlated with lymph node metastasis and TNM staging expression(P<0.05). MRNA translation and protein expression of BRCA2 were inhibited by miR-488-3p. BRCA2 had an inhibitory effect on breast cancer; miR-488-3p can promote the invasion, proliferation, migration and aggregation of MCF7 in breast cancer cells in S phase, and these promoting effects can be inhibited by BRCA2. In nude mice, miR-488-3p promoted the proliferation of MCF7 cells. Conclusion In this study, miR-488-3p inhibiting BRCA2 and promoting breast cancer proliferation may be a new target for breast cancer diagnosis and treatment.

Key words: breast cancer, miR-488-3p, BRCA2