摘要
目的探讨微小RNA-186(miR-186)通过调控Dicer1对乳腺癌细胞MCF-7侵袭转移的影响及其作用机制。方法收集2018年1月至2019年2月于山东省威海市中心医院经手术切除后病理证实为乳腺癌的组织标本及其对应的癌旁正常组织标本各40份,实时荧光定量聚合酶链反应法检测miR-186在乳腺癌组织及对应的癌旁正常组织标本、人乳腺癌细胞株MCF-7以及正常人乳腺上皮细胞MCF10A的表达情况;Transwell法检测miR-186对乳腺癌细胞株MCF-7侵袭能力的影响;划痕实验检测miR-186对乳腺癌细胞株MCF-7迁移能力的影响;生物信息学预测及双荧光素酶报告基因实验验证miR-186和Dicer1的靶向关系;蛋白质印迹法检测Dicer1在各细胞株中的表达。结果乳腺癌组织中miR-186的表达水平显著低于癌旁正常组织[(0. 38±0. 11)比(0. 98±0. 24)](P <0. 05);miR-186在人乳腺癌细胞株MCF-7中的表达水平显著低于正常人乳腺上皮细胞MCF10A[(0. 14±0. 05)比(1. 04±0. 27)](P <0. 05)。过表达miR-186后,MCF-7-mimic-miR-186细胞穿过Matrigel基质胶的数量及覆盖划痕区域少于MCF-7和MCF-7-mimic-NC(均P <0. 05)。经www. microRNA. org网站预测发现miR-186和Dicer1有互补结合序列,双荧光素酶报告实验证实二者的靶向结合关系。人乳腺癌细胞株MCF-7中Dicer1表达水平显著低于正常人乳腺上皮细胞MCF10A,MCF-7-mimic-miR-186中Dicer1表达水平明显高于MCF-7和MCF-7-mimic-NC(均P <0. 05)。结论 miR-186通过靶向正调控Dicer1的表达,抑制人乳腺癌细胞株MCF-7的侵袭迁移能力。
Objective To investigate the effect of microRNA-186( miR-186) on invasion and metastasis of breast cancer cell line MCF-7 by regulating Dicer1. Methods Forty specimens of breast cancer tissue and paracancerous normal tissue determined by surgical and pathological findings were collected from January 2018 to February 2019 in Weihai Central Hospital,Shandong Province. Expressions of miR-186 in breast cancer tissue,paracancerous normal tissue,human breast cancer cell line MCF-7 and human normal breast epithelial cell line MCF10 A were detected by real-time fluorescence quantitative polymerase chain reaction. Invasion ability of MCF-7 cells affected by miR-186 was examined by Transwell assay and cell migration ability was examined by scratch assay. The relation between miR-186 and Dicer1 was verified by bioinformatics software prediction and dual-luciferase reporter gene assay. Expression of Dicer1 was detected by western blotting. Results Breast cancer tissue had lower expression of miR-186 than normal adjacent tissue[( 0. 38 ± 0. 11) vs( 0. 98 ± 0. 24) ]( P < 0. 05). Human breast cancer cells MCF-7 had lower expression of miR-186 than normal breast epithelial cells MCF10 A[( 0. 14 ±0. 05) vs( 1. 04 ± 0. 27) ]( P < 0. 05). The number of MCF-7-mimic-miR-186 cells passing through Matrigel matrix gel and the scratch area were significantly less than those of MCF-7 and MCF-7-mimic-NC( all P < 0. 05).According to the prediction of www. microRNA. org website,miR-186 and Dicer1 had complementary binding sequences,and dual-luciferase reporter gene assay confirmed the targeted binding relation between them. Breast cancer cells MCF-7 showed lower expression of Dicer1 than normal breast epithelial cells MCF10 A( P < 0. 05).MCF-7-mimc-mir-186 cells showed higher expression of Dicer1 than MCF-7 and MCF-7-mimc-NC( both P <0. 05). Conclusion MiR-186 inhibits the invasion and migration of breast cancer cell line MCF-7 by up-regulating Dicer1 expression.
作者
王明蕾
高巧燕
张道强
Wang Minglei;Gao Qiaoyan;Zhang Daoqiang(Logistics Management Center,Weihai Central Hospital,Weihai Key Laboratory of Autoimmunity,Shandong Province,Weihai 264400,China;Department of Nursing,Weihai Central Hospital,Shandong Province,Weihai 264400,China;Central Laboratory,Weihai Central Hospital,Shandong Province,Weihai 264400,China)
出处
《中国医药》
2020年第1期67-70,共4页
China Medicine
基金
国家自然科学基金(21577106)~~
