Abstract：Objective To investigate the expression of microRNA-21 (miR-21) in cervical cancer (CC), and to explore its effect on the proliferation of CC cells and its mechanism.Methods We collected peripheral blood (PB), cancer tissues and paracancerous tissues of 56 CC patients, and 67 healthy volunteers from Inner Mongolia People′s Hospital between January 2016 and June 2018. We also calculated the tumor size, and used real-time quantitative PCR (qRT-PCR) to detect the content of HPV16, HPV18 and miR-21 in PB of CC patients and healthy volunteers, as well as the content of miR-21 in cervical cancer tissues and paracancerous tissues; the correlation between miR-21 with HPV16 and HPV18 was analyzed by Pearson′s correlation analysis; miR-21 expression levels in cervical normal epithelial cells H8, CC cell lines C4-I, C4-II, Hela, SiHa, C33A, Caski, MS751 cells,and HeLa cells after miR-21 inhibitor treatment for 24 h were detected by qRT-PCR;the effect of miR-21 inhibitor on proliferation of H8, Hela, C4-I and Caski cells was detected in CCk-8 assay;the effect of miR-21 inhibitor on the mRNA and protein expression of CyclinD1, CyclinE, CDK4, CDK6 and CYLD in Hela and Caski cells were detected by qRT-PCR and Western blot, respectively.Results The levels of HPV16, HPV18 and miR-21 in PB of CC patients were significantly higher than those in healthy volunteers. miR-21 level was higher in cancer tissues than in paracancerous tissues and PB of CC patients (P<0.05). miR-21 content was positively correlated with HPV16, HPV18 and tumor size (P<0.05). miR-21 level was higher in CC cell lines C4-I, C4-II, Hela, SiHa, C33A, Caski and MS751 cells than that in normal cervical epithelial cells H8. miR-21 inhibitor significantly down-regulated miR-21 expression in Hela and inhibited proliferation of Hela, C4-I and Caski cell (P<0.05), but did not affect normal cervical Epithelial cells H8 (P>0.05). miR-21 inhibitor significantly inhibited the expression of CyclinD1, CyclinE, CDK4 protein and mRNA in Hela and Caski cells (P<0.05), up-regulated the expression of CYLD protein and mRNA (P<0.05). That inhibited both CDK6 protein and mRNA in Caski cells (P<0.05), but had no significant effect on CDK6 protein or mRNA in Hela cells (P>0.05).Conclusion miR-21 content was highly expressed in CC tissues and paracancerous cells, which promotes cell proliferation by inhibiting CYLD protein-mediated cell cycle arrest. It suggests that miR-21 may be involved in the development of CC, which may become a similar diagnostic indicator for CC as HPV16 and HPV18, and participate in tumor growth.
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