Polymorphism effect of Pre-miR-146a on the proliferation of cervical cancer cells
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    Abstract:

    Objective:To transfect the eukaryotic expression plasmid of Pre-miR-146a with a single nucleotide polymorphism (SNP) and observe the expression efficiency of miR-146a in cervical cancer HeLa cells,and then explore the possible mechanism of miRNA-146a on the proliferation of HeLa cells. Methods:We directly cloned a 370-bp fragment including miR-146a precursor from the human gDNA into the pcDNA3.1 (+) plasmid to construct recombinant Pre-miR-146a plasmid by PCR. A Pre-miR-146a-G or Pre-miR-146a-C was transfected into HeLa cell line and the expression of miR-146a was analyzed by real-time PCR. Cell proliferation was measured by cell counting kit-8 assay. Real-time PCR and Western blotting assays were performed to assess the expression of TNF receptor-associated factor 6 (TRAF6). Results:The fragment including miR-146a precursor was successfully cloned into the pcDNA3.1 (+) plasmid and the recombinant expression plasmid Pre-miR-146a-G or Pre-miR-146a-C was constructed. After transfection of Pre-miR-146a-G or Pre-miR-146a-C recombinant plasmid into HeLa cell line,the expression of miR-146a was significantly increased (P < 0.05). The miR-146a expression of the Pre-miR-146a-C plasmid transfected group was higher than that of the Pre-miR-146a-G plasmid group (P < 0.05). After transfection with recombinant plasmid 48 h,the cell viability of HeLa cell line was increased (P < 0.05) and TRAF6 protein expression was down-regulated (P < 0.05),but the mRNA level of TRAF6 did not changed (P > 0.05). Conclusion:The SNP in Pre-miR-146a enhanced the expression of miR-146a and promoted the proliferation of HeLa cells,probably by inhibiting the NF-κB signaling pathway by down-regulating TRAF6.

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胡启明,丁 波,崔毓桂,周德兰,梁佳乐,韩男男,韩素萍. miR-146a前体多态性对宫颈癌细胞增殖的影响[J].南京医科大学学报(自然科学版英文版),2016,(6):676-681.

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History
  • Received:September 22,2015
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  • Online: June 15,2016
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