LncRNA H19 regulates the expression of its target gene HOXA10 in endometrial carcinoma through competing with miR-612.
LncRNA H19 regulates the expression of its target gene HOXA10 in endometrial carcinoma through competing with miR-612.
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DOI：
10.26355/eurrev_201808_15617
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发表时间：
2018-08
影响因子：
3.3
通讯作者：
L. Zhang;D-L Wang;YU P.
中科院分区：
文献类型：
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作者：
L. Zhang;D-L Wang;YU P.
目的 研究长链非编码RNA（lncRNA）H19在子宫内膜癌中的作用，并探讨其作用机制。患者及方法采用定量逆转录聚合酶链反应（qRT-PCR）检测子宫内膜癌中lncRNA H19、miR-612和HOXA10的表达，并分析lncRNA H19与生存时间的关系。通过细胞转染实验完成lncRNA H19在子宫内膜癌细胞中的高表达或敲低。使用细胞计数试剂盒8（CCK-8）检测子宫内膜癌细胞活力的变化。进行双荧光素酶报告基因测定以验证 miR-612 可以与 lncRNA H19 或 HOXA10 结合。采用QRT-PCR和Western Blot法检测lncRNA H19过表达或敲低前后子宫内膜癌细胞中HOXA10表达的变化。结果 子宫内膜癌中lncRNA H19、HOXA10高表达，miR-612低表达。生存曲线表明lncRNA H19与患者生存呈负相关。采用qRT-PCR检测子宫内膜癌细胞HEC1-A、HEC1-B、AN3CA、Ishikawa中lncRNA H19 mRNA的表达量。发现lncRNA H19的表达在AN3CA中最高，在Ishikawa中最低。细胞转染实验使Ishikawa细胞过表达lncRNA H19，AN3CA细胞降低lncRNA H19表达。过表达lncRNA H19后，Ishikawa细胞的活力以及HOXA10的mRNA和蛋白水平增加。然而，敲低lncRNA H19后，AN3CA细胞的活力以及HOXA10的mRNA和蛋白质水平下降。双荧光素酶报告基因检测结果表明 miR-612 可以与 lncRNA H19 和 HOXA10 结合。结论 子宫内膜癌中lncRNA H19的高表达可能通过靶向miR-612来调节其靶基因HOXA10的表达水平,从而促进细胞增殖在子宫内膜癌的发生发展中发挥作用。
OBJECTIVE The role of long non-coding RNA (lncRNA) H19 in endometrial carcinoma was studied, and its mechanism was also explored. PATIENTS AND METHODS Quantitative Reverse Transcriptase-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of lncRNA H19, miR-612, and HOXA10 in endometrial carcinoma, and the relationship between lncRNA H19 and survival time was analyzed. The high expression or knockdown of lncRNA H19 in endometrial cancer cells was completed by cell transfection experiments. Cell counting kit-8 (CCK-8) assay was used to detect changes in the viability of endometrial cancer cells. Dual luciferase reporter assay was performed to verify that miR-612 could bind to lncRNA H19 or HOXA10. QRT-PCR and Western Blot assays were used to detect changes in the expression of HOXA10 in endometrial cancer cells before and after overexpression or knockdown of lncRNA H19. RESULTS The expression of lncRNA H19 and HOXA10 was high in endometrial carcinoma and miR-612 was lowly expressed. The survival curve suggested that lncRNA H19 was negatively correlated with patient survival. The mRNA expression of lncRNA H19 in endometrial cancer cells, including HEC1-A, HEC1-B, AN3CA, and Ishikawa, was detected by qRT-PCR. It was found that the expression of lncRNA H19 was highest in AN3CA and lowest in Ishikawa. The cell transfection experiments allowed Ishikawa cells to overexpress lncRNA H19 and AN3CA cells to reduce lncRNA H19 expression. After overexpression of lncRNA H19, the viability of Ishikawa cells as well as the mRNA and protein levels of HOXA10 increased. However, after knocking down lncRNA H19, the viability of AN3CA cells along with the mRNA and protein levels of HOXA10 decreased. The dual luciferase reporter assay results suggested that miR-612 could bind to lncRNA H19 and HOXA10. CONCLUSIONS The high expression of lncRNA H19 in endometrial carcinoma may regulate the expression level of its target gene HOXA10 by targeting miR-612, thus promoting cell proliferation to play a role in the development of endometrial carcinoma.