长链非编码RNA H19靶向miR-141调控卵巢癌细胞的迁移和侵袭行为

Long-chain non-coding RNA H19 targeting miR-141 regulates migration and invasion of ovarian cancer cells

目的:探讨长链非编码RNA H19靶向miR-141调控卵巢癌细胞的迁移和侵袭行为及其机制。方法:q PCR检测H19和miR-141在卵巢癌组织中的表达情况及H19在不同卵巢癌细胞株中的表达;分析H19和卵巢癌患者的临床病理资料之间的关联;双荧光素酶报告基因检测H19与miR-141之间的相互作用;划痕愈合实验和Transwell侵袭实验检测抑制H19后卵巢癌细胞的迁移和侵袭能力的变化情况,q PCR检测H19和miR-141之间的相互调控的关系;Western blot检测抑制H19后ZEB1蛋白的表达情况。结果:与正常卵巢组织相比,H19在卵巢癌组织中表达上调,miR-141在卵巢癌中的表达水平下调,与其他卵巢癌细胞株相比,ES-2细胞中H19表达最高;H19的表达与卵巢癌的病理分期相关以及淋巴结转移情况有关,随着分期越高,H19在卵巢癌组织中表达越高,淋巴结转移的患者中H19的表达也相对较高;双荧光素酶实验证实H19能与miR-141的3'UTR特异性结合,可以调控miR-141的表达与活性;抑制H19的表达后可以抑制卵巢癌细胞的迁移和侵袭能力;miR-141可以负反馈调节H19的表达,抑制H19和miR-141的表达后,ZEB1蛋白的表达水平受到相应的抑制。结论:H19调控miR-141的表达通过影响ZEB1蛋白的表达参与卵巢癌细胞迁移和侵袭能力的调控。

Objective:To investigate the effect of long-chain non-coding RNA H19 targeting miR-141 on the migration and invasion of ovarian cancer cells and its mechanism.Methods:The expression of H19 and miR-141 in different ovarian cancer cell lines was detected by qPCR.To analyze the association between H19 and clinicopathological data in patients with ovarian cancer.Double luciferase reporter gene was used to detect the interaction between H19 and miR-141.Scaling healing experiment and Transwell invasion test were used to detect the changes of migration and invasion ability of ovarian cancer cells after H19,qPCR detects the interrelationship between H19 and miR-141;Western blot was used to detect the expression of ZEB1 protein after H19.Results:Compared with normal ovarian tissue,the expression of H19 was up-regulated in ovarian cancer and the expression of miR-141 was down-regulated in ovarian cancer.Compared with other ovarian cancer cell lines,the expression of H19 was the highest in ES-2 cells.The expression of H19 was related to the pathological stage of ovarian cancer and the lymph node metastasis.The higher the expression of H19,the higher the expression of H19 in ovarian cancer,and the higher expression of H19 in lymph node metastasis.Double luciferase assay confirmed that H19 could bind to the 3′UTR of miR-141,which could regulate the expression and activity of miR-141.Inhibition of H19 expression could inhibit ovarian cancer cell migration and invasion ability;miR-141 could negatively regulate the expression of H19,inhibit the expression of H19 and miR-141,ZEB1 protein expression levels were inhibited accordingly.Conclusion:The expression of miR-141 by H19 regulates the expression of ZEB1 protein in the regulation of ovarian cancer cell migration and invasion.