Abstract:Objective To investigate the correlation between miR-499 rs3746444 polymorphism and clinical features of cervical cancer, and its effect on the proliferation of cervical cancer cells.Methods Case-control study was conducted among 857 patients with cervical cancer from June 2012 to December 2018 in the Inner Mongolia people's hospital, and 873 healthy volunteers were included as the controls. The age of both groups were 23~62 years old. The polymorphism of miR-499 at rs3746444 was genotyped by TaqMan probe method. And the pGL3- Sox6 was constructed Renilla luciferase vector of GG or AA of 3 ' UTR and miR-499 was inserted into prl-sv40 plasmid. Lipo2000 was used to transfect human cervical epithelial immortalized cells H8 and human cervical cancer cell lines: adenocarcinoma like SiHa, epithelioid HeLa and c33a. Cell proliferation was detected by CCK-8 assay; The luciferase activities of frefly and Renilla were analyzed by double luciferase reporter gene assay, and the protein expressions of cyclin D1, cyclinE, CDK4, Cdk6 and Sox6 in HeLa and SiHa cells were detected by Western blot.Results The risk of cervical cancer in the AG and GG genotypes was 1.09 and 2.41 times higher than that of the AA genotype, respectively. The risk of cervical cancer in individuals carrying the G allele was 1.28 times that of the AA genotype. The proportion of patients at stage III and high differentiation features in cervical cancer patients with GG genotype was significantly higher than that of AA genotype. The different genotypes of rs3746444 had no significant effect on the proliferation of normal cervical H8 cells. The OD450 of Hela, C33A and SiHa cells treated with miR-499 GG+Sox6-3'UTR was significantly lower than that of miR-499 AA+Sox6-3'UTR group at 72h, and higher than that of Sox6-3'UTR group (P<0.05). The Cyclin D1, CyclinE, CDK4 and CDK6 in HeLa and SiHa cells of miR-499 GG+Sox6-3'UTR treatment group were significantly higher than those of miR-499 AA+Sox6-3'UTR group, lower than Sox6-3'UTR group (P<0.05). Compared with the miR-499 AA+Sox6-3'UTR-treated group, the expression level of Sox6 in the miR-499 GG+Sox6-3'UTR-treated group was increased (P<0.05). Both were lower than the Sox6-3'UTR control group (P<0.05).Conclusion The rs3746444 mutation (A>G) of miR-499 is closely related to the growth and differentiation of cervical cancer. It can significantly promote the growth and proliferation of cervical cancer cells by up-regulating the overexpression of CyclinD1 mediated by Sox6.
苏俊玲, 乌云, 杨文静, 张燕. miR-499 rs3746444多态性抑制宫颈癌细胞的增殖作用及分子机制[J]. 中国生育健康杂志, 2021, 32(2): 143-149.
SU Junling, WU Yun, YANG Wenjin, ZHANG Yan. Inhibitory effect of miR-499 rs3746444 polymorphism on proliferation of cervical cancer cells and its molecular mechanism. Chinese Journal of Reproductive Health, 2021, 32(2): 143-149.
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