IKCa1通道对HeLa细胞迁移、侵袭及miRNA表达谱的影响

发布时间：2018-03-03 14:25

  本文选题：宫颈肿瘤　切入点：HeLa细胞　出处：《山东医药》2017年41期 　论文类型：期刊论文

【摘要】：目的探讨钙激活钾离子(IKCa1)通道对宫颈癌He La细胞迁移、侵袭及miRNA表达谱的影响。方法取生长期He La细胞,随机分为观察1~5组及DMSO组,分别给予0、10.0、20.0、30.0、40.0μmol/L TRAM-34及等体积DMSO(TRAM-34溶剂)处理24、48和72 h,用划痕实验检测He La细胞迁移特性;用Transwell侵袭实验检测48 h细胞侵袭特性。用高通量测序法检测IKCa1通道阻断前后miRNA表达谱的差异,并用qRT-PCR法对部分差异表达miRNAs进行验证。运用miRanda软件预测差异miRNA可能调控的靶基因,并对这些靶基因进行通路富集分析。结果划痕实验,TRAM-34干预后He La细胞迁移受抑制,随着TRAM-34浓度增加及作用时间延长,抑制迁移作用越明显(P0.001)。Transwell侵袭实验显示,He La细胞的侵袭率随TRAM-34浓度增加呈递减趋势(F=384.050,P0.001)。通过高通量测序筛选出15个差异有统计学意义而表达上调的miRNA,5个表达下调的miRNA。qRT-PCR结果显示差异性miRNA的表达趋势与测序结果一致。生物信息学分析发现,hsa-miR-143-5p的靶基因在癌症相关通路、p53基因信号通路及MAPK信号通路上出现聚集;而hsa-miR-421的靶基因在肿瘤蛋白多糖、Rap1信号通路和Wnt信号通路出现聚集。结论阻断IKCa1通道可抑制He La细胞迁移和侵袭,影响miRNAs表达谱。
[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.
【作者单位】： 西南医科大学附属医院;
【基金】：四川省卫生厅科研课题(110373)
【分类号】：R737.33
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本文编号：1561413