上调miR-181a和miR-181b表达对HeLa细胞生物学特性的影响

Effects of upregulation of miR-181a and miR-181b on biological properties of HeLa cells

目的:探讨miR-181a和miR-181b表达改变对HeLa细胞生物学特性的影响。方法:收集18例宫颈癌手术标本及18例正常宫颈组织标本,提取总RNA,应用实时荧光定量PCR技术检测miR-181a和miR-181b的表达情况。选取人宫颈癌细胞系HeLa细胞为研究对象,转染miR-181a和miR-181b成熟体。应用实时定量PCR技术检测转染后HeLa细胞中miR-181a和miR-181b的表达情况;流式细胞术检测细胞凋亡和周期变化情况;CCK-8实验及细胞生长曲线检测细胞增殖能力。结果:miR-181a和miR-181b在宫颈癌组织中低表达。转染microRNA成熟体使细胞中miR-181a和miR-181b表达增高;HeLa细胞凋亡增加,细胞增殖能力下降,周期阻滞。结论:上调miR-181a和miR-181b的表达能够促进HeLa细胞凋亡,抑制细胞增殖,导致细胞G0/G1期阻滞。miR-181a和miR-181b可能为宫颈癌临床靶向治疗的靶点选择和药物开发提供理论基础。

Objective：To investigate the effects of the upregulation expression ot mlH - 181 a and miR - 181b on biological properties of HeLa cells. Methods：Tissues from 18 cases of cervical cancer and 18 samples of normal hu-man cervix were collected after surgical operation. Total miRNA was extracted and the relative expression of miR - 181a and miR - 181b were quantified by real -time quantitative PCR. microRNA mimics were transfected into cervi-cal cancer HeLa cells. The expressions of miR-181a and miR-181b were determined by real-time quantitative PCR; The changes of cell apoptosis and cycle were measured by flow cytometry. The cell counting Kit-8 （CCK -8 ） assay and growing curve were used to assess the effects miR - 181a and miR - 181b on cell proliferation. Results： miR-181a and miR-181b were markedly down-regulated in cervical cancer comparing with normal cervical sam-ples. Ecotopic expression of miR-181a and miR-181b promoted cell apoptosis, proliferation inhibition and cell cy-cle arrest. Conclusion：miR-181a and miR-181b may be as a tumor suppressor by inducing apoptosis,inhabiting proliferation and blocking cell cycle. Moreover,the miR - 181a and miR - 181b may provide an important theory basis in target choose for cervical carcinoma clinical treatment and pharmaprojects.