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Archive > Volume 24 Issue 7 > 2017,24(7):742-747. DOI:10.3872/j.issn.1007-385X.2017.07.008 Prev Next
Basic Research
Effect of miR-134-5p on proliferation and apoptosis of cervical carcinoma cell and its molecular mechanism
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Abstract:
Objective: To observe effect of miR-134-5p transfection on proliferation and apoptosis of cervical carci-noma cell and to verify its possible molecular mechanism. Methods: Eight pairs of cervical cancer and para-cancer-ous tissuesfrom the patients with cervical cancer who hospitalized in Center of Oncology, Renmin Hospital, Hubei University of Medicine during May to August 2016 were collected. miR-134-5p mimics were transfected into cervi-cal carcinoma Hela and SiHa cells by lipofectomin 2000. MTT and colony formation assays were used to detect pro-liferation of cells. Flow cytometry (FCM）assay was used to test cell cycles and apoptosis of cells. Expressions of miR-134-5p mRNA in cervical carcinoma tissue and cell, and expression of EGFR mRNA in cervical carcinoma cell were detected by qRT-PCR assay. Expressions of EGFR pathway-related proteins in cervical carcinoma cell were examed by Western blotting assay. Results: Expression of miR-134-5p mRNA in cervical carcinoma tissue was significantly lower than that in para-carcinoma tissue (P<0.01). Comparing with the Hela and SiHa cells that transfected with miR-NC, expressions of miR-134-5p mRNA in the Hela and SiHa cells that transfected with miR-134-5p mimics were obviously increased, proliferation abilities of the cells significantly reduced (at the 5th day of the transfection, Hela cell:1.06 ± 0.13 vs 1.32 ± 0.07; SiHa cell: 1.12 ± 0.10 vs 1.42 ± 0.12, all P<0.05), apoptosis rates of the cells obviously increased (Hela cell: [26.53 ± 13.48]% vs [3.25 ± 1.74]%; SiHa cell: [30.49 ± 12.04]% vs [5.12 ±2.86]%, all P<0.05), number of formed colony decreased, ratio of G0/G1 phase cells increased, ratio of the cells in Sand G2/M phase decreased, apoptosis rate of the cells enhanced (all P<0.05), expressions of EGFR mRNA and EG-FR protein in the cells were remarkably down-regulated, among which EGFR mRNA in the the Hela cell down 58%(P<0.01) and in the the SiHa cell down 41% (P<0.05), expressions of downstream target protein for EGFR, p-AKT,p-ERK/2 and Cyclin D1, as well as pEGFR proteins were evidently down-regulated. Conclusion: miR-134-5p couldsignificantly inhibit proliferation of the cervical carcinoma cells and promote their apoptosis, of which possible mo-lecular mechanism might be inhibit activation of EGFR pathway through inhibiting expression of EGFR gene.
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Project Supported:
Project supported by the Scientific Technology Research Foundation of Hubei Provincial Education Department (No. B2016139) and Innovative Foundation for Excellent Young and Middle-aged Talents in Universities of Hubei Province (No. T201510)
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