miR-150靶向调控FOXO4促进宫颈癌C33A细胞生长与生存
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南充市科学技术和知识产权局基金项目


microRNA150 promotes cervical cancer cell C-33A growth and survival by targeting FOXO4
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    摘要:

    【摘要】 目的 检测miR150在宫颈癌细胞C33A中的表达,并探讨其是否通过调控靶向基因FOXO4促进宫颈癌细胞的增殖与凋亡。方法 将携带miR150 mimic(模拟物)和miR150 inhibitor(抑制物)、阴性对照siRNA经 LipofectamineTM 2000 转染至C33A细胞中,将细胞分为 NC组( 阴性对照组) 、miR150 模拟物组( 转染miR150 mimic细胞组) 及 miR150in 抑制物组( 转染miR150 inhibitor细胞组)。采用流式细胞仪、TUNEL检测法、荧光定量PCR、Western bolt 分别检测miR150对宫颈癌C33A细胞周期、凋亡以及周期和凋亡相关基因的影响。3’端非翻译区(UTR)荧光素酶活性分析证实miR150的直接靶向作用。结果 miR150模拟物促进宫颈癌细胞C33A的增殖与凋亡,抑制物作用相反,miR150模拟物促进细胞由G1/G0期进展到S期,从而促进宫颈癌细胞增殖,抑制物相反,miR150调控几个细胞周期、凋亡相关基因CyclinD1、 p27、 BIM和FASL的表达; miR150通过靶向结合FOXO4的3’UTR区从而抑制FOXO4的表达。结论 miR150靶向作用于FOXO4从而调节多种基因的表达以致宫颈癌细胞C33A的增殖与凋亡。

    Abstract:

    【Abstract】 Objective To detect the expression of miR150 in cervical cancer cells C33A and investigate whether it promotes the proliferation and invasion of cervical cancer cells by regulating the target gene FOXO4.Methods The effects of miR150 on cell cycle and apoptosis, as well as the expression of cycleand apoptosisrelated genes, were determined using flow cytometry, TUNEL assay, qRTPCR, and Western blot, respectively. The direct target of miR150 was confirmed using 3' untranslated region (UTR) luciferase reporter assay.Results miR150 promotes cervical cancer cell C33A survival and growth, while the inhibition of miR150 suppresses these actions. miR150 also induced the cell cycle progressionfrom G1/G0 to S phase, resulting in an enhancement of growth. Several cell cycleand apoptosisrelated genes, CyclinD1, p27, BIM, and FASL were modulated by miR150.Moreover, miR150 directly reduced the expression of FOXO4, which regulates the expression of CyclinD1, p27, BIM, and FASL, by targeting its 3' UTR.Conclusion Our data demonstrate that elevated miR150 targets FOXO4 expression and therefore regulates multiple genes expression, resulting in cervical cancer cell C33A growth and survival.

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  • 在线发布日期: 2019-09-10
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