摘要
目的研究miR-613靶向调控高尔基体磷蛋白3(GOLPH3)基因对乳腺癌MCF-7细胞的增殖、迁移和侵袭的机制。方法用脂质体技术分别将miR-NC、miR-613 mimic、anti-miR-NC、anti-miR-613、si-NC、si-GOLPH3、miR-613 mimic与pcDNA、miR-613mimic与pcDNA-GOLPH3转染至MCF-7细胞。用实时荧光定量聚合酶链反应检测miR-613和GOLPH3 mRNA的表达,用噻唑蓝法、Transwell法检测MCF-7细胞的增殖、迁移和侵袭。结果HBL^-100组和MCF-7组的miR-613表达量分别为1.03±0.09和0.37±0.04,GOLPH3 mRNA表达量分别为1.07±0.11和3.25±0.26,GOLPH3蛋白表达量分别为0.34±0.05和0.75±0.08,MCF-7组的上述指标与HBL^-100组比较,差异均有统计学意义(均P<0.05)。miR-NC组和miR-613组72 h时细胞增殖率分别为(0.97±0.09)%和(0.63±0.06)%,迁移细胞数分别为(86.79±8.13)和(40.35±5.27)个,侵袭细胞数分别为(68.43±6.77)和(31.54±4.68)个,miR-613组的上述指标与miR-NC组比较,差异均有统计学意义(均P<0.05)。双荧光素酶报告基因实验证实miR-613能够靶向结合GOLPH3。结论过表达miR-613可通过靶向调控GOLPH3抑制MCF-7细胞的增殖、迁移和侵袭。
Objective To investigate the effects of miR-613 targeting golgi phosphoprotein 3(GOLPH3)gene on the proliferation,migration and invasion of MCF-7 cells in breast cancer and its mechanism.Methods Using liposome technology,miR-NC,miR-613 mimic,anti-miR-NC,anti-miR-613 group,si-NC,si-GOLPH3,miR-613 mimic and pcDNA,miR-613 mimic and pcDNA-GOLPH3 were transfected into MCF-7 cells.Reverse transcription-polymerase chain reaction was used to detect the expressions of miR-613 and GOLPH3 mRNA.Methyl thiazolyl tetrazolium method and Transwell method were used to detect the proliferation,migration and invasion of MCF-7 cells.Results The expression levels of miR-613 in the HBL^-100 group and the MCF-7 group were 1.03±0.09 and 0.37±0.04,the GOLPH3 mRNA expressions were 1.07±0.11 and 3.25±0.26,and the GOLPH3 protein expression were 0.34±0.05 and 0.75±0.08.The above indexes in MCF-7 group were significantly different from those in HBL^-100 group(all P<0.05).The cell proliferation rates of miR-NC group and miR-613 group at 72 h were(0.97±0.09)%and(0.63±0.06)%,respectively;the number of migrating cells were(86.79±8.13)and(40.35±5.27),the number of invading cells were(68.43±6.77)and(31.54±4.68).The above indexes in miR-613 group were significantly different from those in miR-NC group(all P<0.05).Double luciferase reporter gene experiments confirmed that miR-613 can target GOLPH3.Conclusion Overexpression of miR-613 can inhibit the proliferation,migration and invasion of MCF-7 cells through targeted regulation of GOLPH3.
作者
周春
王光华
张帮柱
余峰彬
范耀刚
ZHOU Chun;WANG Guang-hua;ZHANG Bang-zhu;YU Feng-bin;FAN Yao-gang(Department of Surgery,School of Clinical Medicine,Panzhihua University,Panzhihua 617000,Sichuan Province,China;Department of Hepatology,Panzhihua Central Hospital,Panzhihua 617067,Sichuan Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2020年第19期3066-3069,3074,共5页
The Chinese Journal of Clinical Pharmacology
基金
四川省教育厅课题资助项目(16ZB0480)。