本文關(guān)鍵詞： 微小RNA- 叉頭盒蛋白M 肝細(xì)胞癌 細(xì)胞增殖　出處：《第三軍醫(yī)大學(xué)學(xué)報》2017年01期 　論文類型：期刊論文

【摘要】：目的探討miR-134下調(diào)叉頭盒蛋白M1(forkhead box M1,FOXM1)的機(jī)制及其影響肝細(xì)胞癌(hepatocellular carcinoma,HCC)細(xì)胞增殖的作用。方法應(yīng)用CCK-8法檢測miR-134對HCC細(xì)胞增殖的作用;經(jīng)Western blot和Real-time PCR檢測人正常肝細(xì)胞L02和5種HCC細(xì)胞系中FOXM1和miR-134的表達(dá);用miR-134模擬物(miR-134 mimic)或miR-134抑制劑(miR-134 inhibitor)轉(zhuǎn)染HCC細(xì)胞后,經(jīng)Western blot和Real-time PCR檢測FOXM1及其下游靶基因Cyclin D1的表達(dá);應(yīng)用生物信息學(xué)分析miR-134在人FOXM1 3'-UTR的可能結(jié)合位點(diǎn),通過報告基因檢測實(shí)驗(yàn)分析miR-134與FOXM1的3'-UTR的特異性結(jié)合;將miR-134 mimic和FOXM1表達(dá)質(zhì)粒(無3'-UTR)共轉(zhuǎn)染于HCC細(xì)胞后,經(jīng)CCK-8法檢測細(xì)胞的增殖情況。結(jié)果 miR-134可顯著抑制HCC細(xì)胞增殖(P0.05);與人正常肝細(xì)胞L02相比,miR-134在HCC細(xì)胞中的表達(dá)明顯降低(P0.05),而FOXM1蛋白表達(dá)明顯增強(qiáng),二者存在負(fù)相關(guān)(R2=0.705,P0.05);miR-134可顯著下調(diào)FOXM1蛋白及其下游靶基因Cyclin D1的表達(dá)(P0.05);報告基因檢測實(shí)驗(yàn)證實(shí)miR-134可特異性結(jié)合于FOXM1 mRNA的3'-UTR(P0.01);細(xì)胞增殖實(shí)驗(yàn)檢測結(jié)果顯示,過表達(dá)缺失3'-UTR的FOXM1可顯著減弱miR-134對HCC細(xì)胞增殖的抑制效應(yīng)(P0.05)。結(jié)論 miR-134通過靶向結(jié)合于FOXM1的3'-UTR而下調(diào)FOXM1蛋白的表達(dá),從而抑制HCC細(xì)胞的增殖。
[Abstract]:Objective to investigate the down-regulation of forkhead box M1 by miR-134. FOXM1) and its effect on hepatocellular carcinoma. Methods CCK-8 assay was used to detect the effect of miR-134 on the proliferation of HCC cells. The expression of FOXM1 and miR-134 in L02 and 5 HCC cell lines were detected by Western blot and Real-time PCR. HCC cells were transfected with miR-134 mimic or miR-134 inhibitor HCC. The expression of FOXM1 and its downstream target gene Cyclin D1 was detected by Western blot and Real-time PCR. Bioinformatics was used to analyze the possible binding sites of miR-134 in human FOXM1 3- UTR. The specific binding of miR-134 to FOXM1 was analyzed by reporter gene detection. MiR-134 mimic and FOXM1 expression plasmids were co-transfected into HCC cells. The proliferation of HCC cells was detected by CCK-8. Results miR-134 could significantly inhibit the proliferation of HCC cells. Compared with human normal hepatocytes L02, the expression of miR-134 in HCC cells was significantly decreased, while the expression of FOXM1 protein was significantly increased. There was a negative correlation between them. MiR-134 could significantly down-regulate the expression of FOXM1 protein and its downstream target gene Cyclin D1. The reporter gene assay confirmed that miR-134 could specifically bind to FOXM1 mRNA. The results of cell proliferation test showed that. Overexpression of FOXM1 with deletion of 3H-UTR significantly attenuated the inhibitory effect of miR-134 on the proliferation of HCC cells (P0.05). Conclusion miR-134 can down-regulate the expression of FOXM1 protein by targeting the 3- UTR binding to FOXM1. Thus, the proliferation of HCC cells was inhibited.
【作者單位】： 第三軍醫(yī)大學(xué)基礎(chǔ)醫(yī)學(xué)部生物化學(xué)與分子生物學(xué)教研室;
【基金】：國家自然科學(xué)基金面上項目(81672377,31470066)~~
【分類號】：R735.7
【正文快照】： 生物化學(xué)與分子生物學(xué)教研室)specific binding of miR-134 with 3'-UTR of FOXM1 mRNA.After co-transfected with miR-134 mimic orFOXM1-expressing plasmids(without 3'-UTR),the HCC cell proliferation was detected by CCK-8 assay.Results miR-134 dramatically suppr，

本文編號：1484550