發(fā)布時(shí)間：2018-03-03 14:25

  本文選題：宮頸腫瘤　切入點(diǎn)：HeLa細(xì)胞　出處：《山東醫(yī)藥》2017年41期 　論文類型：期刊論文

【摘要】：目的探討鈣激活鉀離子(IKCa1)通道對(duì)宮頸癌He La細(xì)胞遷移、侵襲及miRNA表達(dá)譜的影響。方法取生長(zhǎng)期He La細(xì)胞,隨機(jī)分為觀察1~5組及DMSO組,分別給予0、10.0、20.0、30.0、40.0μmol/L TRAM-34及等體積DMSO(TRAM-34溶劑)處理24、48和72 h,用劃痕實(shí)驗(yàn)檢測(cè)He La細(xì)胞遷移特性;用Transwell侵襲實(shí)驗(yàn)檢測(cè)48 h細(xì)胞侵襲特性。用高通量測(cè)序法檢測(cè)IKCa1通道阻斷前后miRNA表達(dá)譜的差異,并用qRT-PCR法對(duì)部分差異表達(dá)miRNAs進(jìn)行驗(yàn)證。運(yùn)用miRanda軟件預(yù)測(cè)差異miRNA可能調(diào)控的靶基因,并對(duì)這些靶基因進(jìn)行通路富集分析。結(jié)果劃痕實(shí)驗(yàn),TRAM-34干預(yù)后He La細(xì)胞遷移受抑制,隨著TRAM-34濃度增加及作用時(shí)間延長(zhǎng),抑制遷移作用越明顯(P0.001)。Transwell侵襲實(shí)驗(yàn)顯示,He La細(xì)胞的侵襲率隨TRAM-34濃度增加呈遞減趨勢(shì)(F=384.050,P0.001)。通過(guò)高通量測(cè)序篩選出15個(gè)差異有統(tǒng)計(jì)學(xué)意義而表達(dá)上調(diào)的miRNA,5個(gè)表達(dá)下調(diào)的miRNA。qRT-PCR結(jié)果顯示差異性miRNA的表達(dá)趨勢(shì)與測(cè)序結(jié)果一致。生物信息學(xué)分析發(fā)現(xiàn),hsa-miR-143-5p的靶基因在癌癥相關(guān)通路、p53基因信號(hào)通路及MAPK信號(hào)通路上出現(xiàn)聚集;而hsa-miR-421的靶基因在腫瘤蛋白多糖、Rap1信號(hào)通路和Wnt信號(hào)通路出現(xiàn)聚集。結(jié)論阻斷IKCa1通道可抑制He La細(xì)胞遷移和侵襲,影響miRNAs表達(dá)譜。
[Abstract]:Objective to investigate the effects of Ca ~ (2 +) -activated potassium ion Ike _ (Ca _ 1) channel on migration, invasion and miRNA expression profile of cervical cancer Hela cells. The migration characteristics of He-La cells were detected by scratch test at 2448 h and 72 h after treatment with 40 渭 mol/L TRAM-34 and DMSO(TRAM-34 solvent of equal volume. The invasion characteristics of He-La cells were detected by Transwell invasion assay. The differences of miRNA expression profiles before and after IKCa1 channel blocking were detected by high-throughput sequencing. Some differentially expressed miRNAs were verified by qRT-PCR method. The target genes regulated by differential miRNA were predicted by miRanda software, and these target genes were enriched and analyzed. Results the migration of He-La cells was inhibited after TRAM-34 intervention. With the increase of TRAM-34 concentration and the prolongation of the action time, The more obvious the inhibitory effect on migration was, the more obvious the invasion rate of He-La cells showed a decreasing trend with the increase of TRAM-34 concentration. Fifteen differentially expressed miRNAs and 5 down-regulated miRNA.qRT-PCR were screened by high-throughput sequencing. The results showed that the trend of differential miRNA expression was consistent with that of sequencing. Bioinformatics analysis showed that the target genes of hsa-miR-143-5p were clustered in p53 gene signaling pathway and MAPK signaling pathway in cancer related pathway. However, the target genes of hsa-miR-421 were clustered in the tumor proteoglycans Rap1 signaling pathway and Wnt signaling pathway. Conclusion blocking the IKCa1 channel can inhibit the migration and invasion of He-La cells and affect the miRNAs expression profile.
【作者單位】： 西南醫(yī)科大學(xué)附屬醫(yī)院;
【基金】：四川省衛(wèi)生廳科研課題(110373)
【分類號(hào)】：R737.33
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本文編號(hào)：1561413