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目的探索长链非编码RNA(lncRNA)肺腺癌转移相关转录子1(metastasis associated lung adenocarcinoma transcript 1,MALAT1)在肝细胞癌(hepatocellular carcinoma,HCC;简称肝癌)细胞中是否可作为竞争性内源RNA(competing endogenous RNA,ceRNA)发挥促迁移功能。方法在Hep G2细胞中通过瞬时转染MALAT1的小干扰RNA(small interfering RNA,siRNA)敲低MALAT1的表达;利用Transwell实验检测敲低MALAT1后对细胞迁移能力的影响;利用RNA pull-down实验确认MALAT1是否可直接吸附miR-126*;荧光定量逆转录PCR(quantitative reverse transcription PCR,qRT-PCR)和Western印迹检测敲低MALAT1后miR-126*靶基因的mRNA和蛋白水平的变化;利用公共数据集癌症基因组图谱(The Cancer Genome Atlas,TCGA)分析MALAT1和miR-126*在肝癌组织样本中的表达情况和预后价值。结果与对照组相比,敲低MALAT1可显著抑制肝癌细胞的迁移能力;MALAT1含有3个miR-126*的结合位点,其野生型序列相对于突变体序列可显著吸附miR-126*;敲低MALAT1后,miR-126*的靶基因CXCL12的mRNA和蛋白水平均显著下调;TCGA肝癌数据分析显示MALAT1在癌组织中显著上调表达,且MALAT1高表达组有较低的无病生存率,而miR-126*与其相反。结论 MALAT1可吸附miR-126*,上调CXCL12的表达,进而促进肝癌的转移。
Objective To investigate whether long-chain non-coding RNA (lncRNA) metastasis associated lung adenocarcinoma transcript 1 (MALAT1) can be used as a competitive endogenous gene in hepatocellular carcinoma (HCC) cells RNA (competing endogenous RNA, ceRNA) play a role in promoting migration. Methods The expression of MALAT1 was knocked down by transient transfection of MALAT1 small interfering RNA (siRNA) in Hep G2 cells. The effect of knocking down MALAT1 on the migration ability was detected by Transwell assay. The RNA pull-down assay was used to confirm MALAT1 can directly adsorb miR-126 *; quantitative reverse transcription PCR (qRT-PCR) and Western blotting were used to detect the mRNA and protein levels of miR-126 * target gene knocked down by MALAT1; The Cancer Genome Atlas (TCGA) was used to analyze the expression and prognostic value of MALAT1 and miR-126 * in HCC tissues. Results Compared with the control group, knockdown of MALAT1 significantly inhibited the migration ability of HCC cells. MALAT1 contained three binding sites of miR-126 *, and its wild-type sequence could significantly adsorb miR-126 * relative to the mutant sequence. The mRNA and protein levels of CXCL12, a target gene of miR-126 *, were significantly down-regulated after low MALAT1. The analysis of TCGA liver cancer data showed that MALAT1 was up-regulated in cancer tissues and the MALAT1-overexpression group had a lower disease-free survival rate miR-126 * is the opposite. Conclusion MALAT1 can up-regulate the expression of CXCL12 and up-regulate the expression of miR-126 *, thereby promoting the metastasis of HCC.