miR-198靶向MAPK1调控宫颈癌HeLa细胞增殖、凋亡和侵袭

目的 探究miR-198对宫颈癌细胞增殖、凋亡和侵袭的作用及其机制。方法 用miR-198 mimic转染HeLa细胞，qRT-PCR检测miR-198和丝裂原活化蛋白激酶1 （Mitogen-activated protein kinase1, MAPK1）的mRNA水平；荧光素酶实验检测miR-198和MAPK1的靶向关系；用pcDNA3.0-MAPK1（pc-MAPK1）和miR-198 mimic转染细胞，CCK-8检测细胞增殖活性，流式细胞术检测细胞凋亡，Transwell法检测细胞侵袭能力，Western blot检测蛋白的表达。结果 miR-198 mimic转染细胞后，miR-198表达水平明显升高，MAPK1 mRNA表达水平明显降低；荧光素酶报告实验表明miR-198序列上存在MAPK1结合位点；miR-198过表达能显著降低宫颈癌细胞增殖倍数、诱导细胞凋亡，同时还能明显降低HeLa细胞侵袭能力；此外，miR-198 mimic能显著抑制MAPK1下游基因核糖体S6激酶2（Ribosomal S6 kinase 2, RSK2）、c-Myc和c-fos的蛋白表达；pc-MAPK1能明显减弱miR-198 mimic对HeLa细胞增殖、凋亡、侵袭及对MAPK1下游蛋白表达的调控作用。结论 miR-198过表达能通过靶向MAPK1抑制宫颈癌细胞的增殖和侵袭能力并诱导细胞凋亡。

miR-198 Targets MAPK1 to Regulate Proliferation,Apoptosis and Invasion of Cervical Cancer HeLa Cells

Objective To investigate the effects and mechanisms of miR-198 on the proliferation, apoptosis and invasion of cervical cancer cells. Methods HeLa cells were transfected with miR-198 mimic and the mRNA levels between miR-198 and mitogen-activated protein kinase 1(MAPK1) were measured by qRTPCR. The relationship of miR-198 and MAPK1 was determined by luciferase reporter assay. pcDNA3.0- MAPK1 (pc-MAPK1) and miR-198 mimic transfection was performed, and cell proliferation, apoptosis and invasion abilities were measured by CCK-8 assay, flow cytometry and Transwell assay, respectively. Western blot was performed for protein levels. Results The level of miR-198 was increased and the mRNA level of MAPK1 was decreased by miR-198 mimic. The result of luciferase reporter assay indicated that there was binding site of MAPK1 on miR-198. Overexpression of miR-198 significantly inhibited the proliferation and invasion, and induced the apoptosis of HeLa cells. In addition, miR-198 mimic down-regulated the expressions of ribosomal S6 kinase 2 (RSK2), c-Myc and c-fos. pc-MAPK1 attenuated the effects of miR-198 mimic on cell proliferation, apoptosis, invasion and downstream proteins expression of MAPK1 in HeLa cells. Conclusion Overexpression of miR-198 inhibits the proliferation, invasion and induces the apoptosis of cervical cancer cells through targeting MAPK1.