Zhang Jing, Wang Lu, Li Baoli, Huo Manpeng, Mu Mingtao, Liu Junjun, Han Jiming
Department of Clinical Medicine, Medical College of Yan'an University, Yan'an, Shaanxi 716000, P.R. China.
Exp Ther Med. 2014 Aug;8(2):591-594. doi: 10.3892/etm.2014.1765. Epub 2014 Jun 6.
The present study aimed to investigate the effect of the inhibition of miR-145 on cyclin-dependent protein kinase 6 (CDK6) and the proliferation of human cervical carcinoma cells. The miR-145 sequence was synthesized and cloned into pcDNA™6.2-GW to construct the recombinant plasmid pcDNA6.2-GW-miR-145. HeLa cells were divided into the micro (mi)R-145, normal control and blank groups. The transcription levels of miR-145 and CDK6 were detected using quantitative polymerase chin reaction and western blot analysis was used to examine the CDK6 protein expression. In addition, the inhibitory effect of miR-145 on the proliferation of HeLa cells was measured by an MTT assay. The recombinant plasmid pcDNA6.2-GW-miR-145 was successfully constructed and used to transfect the HeLa cells in the MiR-145 group. The miR-145 expression level in the miR-145 group was significantly higher than that in the blank group. The CDK6 expression level in miR-145 group was significantly lower than that in the blank group. Furthermore, miR-145 inhibited the proliferation of HeLa cells. In conclusion, miR-145 overexpression suppresses the expression of CDK6 and inhibits the proliferative ability of HeLa cells.
本研究旨在探讨抑制miR-145对细胞周期蛋白依赖性蛋白激酶6（CDK6）及人宫颈癌细胞增殖的影响。合成miR-145序列并将其克隆至pcDNA™6.2-GW中，构建重组质粒pcDNA6.2-GW-miR-145。将HeLa细胞分为微小（mi）R-145组、正常对照组和空白组。采用定量聚合酶链反应检测miR-145和CDK6的转录水平，并用蛋白质印迹分析检测CDK6蛋白表达。此外，通过MTT法检测miR-145对HeLa细胞增殖的抑制作用。成功构建重组质粒pcDNA6.2-GW-miR-145并用于转染MiR-145组的HeLa细胞。MiR-145组中miR-145表达水平显著高于空白组。miR-145组中CDK6表达水平显著低于空白组。此外，miR-145抑制HeLa细胞增殖。总之，miR-145过表达抑制CDK6表达并抑制HeLa细胞的增殖能力。
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