本文選題：MicroRNA--p　切入點(diǎn)：神經(jīng)膠質(zhì)瘤　出處：《中國(guó)病理生理雜志》2017年07期 　論文類型：期刊論文

【摘要】：目的:探討microRNA(miRNA)-483-3p對(duì)人神經(jīng)膠質(zhì)瘤細(xì)胞A172生長(zhǎng)和遷移能力的影響及潛在作用機(jī)制。方法:實(shí)時(shí)熒光定量聚合酶鏈?zhǔn)椒磻?yīng)(RT-q PCR)檢測(cè)人腎胚細(xì)胞系HEK-293和不同神經(jīng)膠質(zhì)瘤細(xì)胞株(A172、U251和SHG44)中miRNA-483-3p的表達(dá)水平。轉(zhuǎn)染miRNA-483-3p抑制序列(miRNA-483-3p inhibitor)下調(diào)A172細(xì)胞中miRNA-483-3p的表達(dá),采用CCK-8法和流式細(xì)胞術(shù)檢測(cè)細(xì)胞活力和周期分布;Transwell實(shí)驗(yàn)檢測(cè)細(xì)胞的遷移;Western blot檢測(cè)周期相關(guān)調(diào)控因子及上皮-間充質(zhì)轉(zhuǎn)化相關(guān)蛋白的水平。雙螢光素酶報(bào)告基因分析法預(yù)測(cè)及驗(yàn)證其可能的靶基因。結(jié)果:miRNA-483-3p在各型神經(jīng)膠質(zhì)瘤細(xì)胞中高表達(dá)。沉默miRNA-483-3p后,A172細(xì)胞的活力下降并呈現(xiàn)出明顯的周期阻滯,且細(xì)胞遷移率也顯著降低。同時(shí)細(xì)胞中cyclin D1、周期蛋白依賴性激酶4、磷酸化視網(wǎng)膜母細(xì)胞瘤蛋白、N-cadherin及vimentin的蛋白表達(dá)水平均顯著降低,E-cadherin和β-catenin的蛋白表達(dá)水平顯著升高。雙螢光素酶報(bào)告基因分析顯示Smad4是miRNA-483-3p的可能作用靶點(diǎn),A172細(xì)胞共轉(zhuǎn)染miRNA-483-3p inhibitor和Smad4 siRNA可部分逆轉(zhuǎn)miRNA-483-3p介導(dǎo)的細(xì)胞增殖及遷移抑制。結(jié)論:沉默miRNA-483-3p可通過(guò)靶向Smad4抑制神經(jīng)膠質(zhì)瘤細(xì)胞株A172的生長(zhǎng)及遷移。
[Abstract]:Objective: to investigate the effect of microRNA(miRNA)-483-3p on the growth and migration of human glioma cell line A172 and its potential mechanism. Methods: Real-time fluorescent quantitative polymerase chain reaction (RT-q PCR) was used to detect human renal embryo cell line HEK-293 and glioma fine cells. The expression level of miRNA-483-3p in A172 U251 and SHG44) was down-regulated by transfection of miRNA-483-3p inhibitory sequence (miRNA-483-3p inhibitor), and the expression of miRNA-483-3p in A172 cells was down-regulated by transfection of miRNA-483-3p inhibitor. CCK-8 assay and flow cytometry were used to detect cell viability and cell cycle distribution. Western blot was used to detect the levels of cycle related regulatory factors and epithelial-mesenchymal transformation related proteins. Results: miRNA-483-3p was highly expressed in glioma cells. After silencing miRNA-483-3p, the activity of A172 cells decreased and the cell cycle arrest was observed. At the same time, the expression of cyclin D1, cyclin dependent kinase 4, phosphorylated retinoblastoma protein N-cadherin and vimentin decreased significantly, and the expression of E-cadherin and 尾 -catenin increased significantly. Double luciferase reporter gene analysis showed that Smad4 is a possible target of miRNA-483-3p in miRNA-483-3p inhibitor and Smad4 siRNA cotransfection with miRNA-483-3p inhibitor and Smad4 siRNA can partially reverse miRNA-483-3p mediated cell proliferation and migration inhibition. Conclusion: silencing miRNA-483-3p can inhibit nerve by targeting Smad4. Growth and migration of glioma cell line A172.
【作者單位】： 鄭州大學(xué)附屬腫瘤醫(yī)院神經(jīng)外科;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(No.81372269)
【分類號(hào)】：R739.41

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