长链非编码RNA H19对口腔癌细胞的迁移和侵袭的影响以及分子机制

Effect of long chain non-coding RNA H19 on the migration and invasion of oral cancer cells and its molecular mechanism

目的探讨长链非编码RNA(lncRNA)H19对口腔癌细胞侵袭、迁移的影响以及作用机制。方法荧光定量聚合酶链反应检测永生化口腔上皮细胞系HIOEC细胞以及口腔癌细胞系CAL27中H19、miR-107、细胞周期蛋白依赖性激酶6(CDK6)的表达。siRNA H19、miR-107 mimics、pcDNA H19、anti-miR-107转染CAL27细胞,Transwell法检测H19和miR-107对细胞侵袭、迁移的影响,TargetScan数据库预测H19、miR-107、CDK6的靶向关系,双荧光素酶报告基因检测H19、miR-107、CDK6的相互作用,蛋白质印迹法(Western blot)检测H19和miR-107对靶基因CDK6蛋白水平的影响。结果H19在CAL27细胞中的表达高于HIOEC细胞(P<0.05)。siRNAH19转染后,H19的表达降低,抑制CAL27细胞的侵袭、迁移能力(P<0.05)。H19与miR-107的3’-UTR特异性结合,调控miR-107的表达活性。miR-107在CAL细胞中的表达低于HIOEC细胞(P<0.05),siRNA H19转染后,miR-107的表达升高,antimiR-107共转染可促进siRNAH19对CAL27细胞的侵袭、迁移能力(P<0.05)。CDK6在CAL27细胞中的表达高于HIOEC细胞(P<0.05),CDK6的表达可被H19和miR-107共同调控。结论lncRNAH19通过靶向调控miR-107/CDK6信号轴影响口腔癌细胞的侵袭、迁移能力,在口腔癌的发生发展过程中起着重要作用。

Objective To investigate the effect of the long chain non-coding RNA H19(lncRNA H19)on the invasion and migration of oral cancer cells and its related molecular mechanism.Methods The expression levels of lncRNA H19,mi R-107,and cyclin-dependent kinase 6(CDK6)in the immortalized oral epithelial cell line HIOEC and the oral cancer cell line CAL27 were detected by real-time quantitative polymerase chain reaction.CAL27 cells were transfected with siRNA H19,mi R-107 mimics,pcDNA H19,or anti-miR-107,and the effects of H19 and miR-107 on the invasion and migration of cells were examined via Transwell assay.The TargetScan database predicted the targeting of H19,miR-107,and CDK6.Double luciferase reporter gene assay was performed to detect interactions among H19,miR-107,and CDK6.Western blot analysis was conducted to examine the effects of H19 and miR-107 on the protein level of the target gene CDK6.Results Compared with that in HIOEC cells,the expression of H19 was significantly increased in CAL27 cells(P<0.05).After transfection with siRNA H19,the expression of H19 decreased,and the invasion and migration ability of CAL27 cells were inhibited(P<0.05).H19 could bind specifically to the 3'-UTR of miR-107 to modulate the expression of miR-107.Compared with that in HIOEC cells,the expression of miR-107 significantly decreased in CAL27 cells(P<0.05).The expression of miR-107 increased after transfection with siRNA H19,and anti-mir-107 cotransfection could promote the invasion and migration ability of siRNA H19 in CAL27 cells(P<0.05).Compared with that in HIOEC cells,CDK6 expression significantly increased in CAL27 cells(P<0.05),and the expression level of the gene was co-regulated by H19 and miR-107(P<0.05).Conclusion lncRNA H19 plays an important role in the development of oral cancer.It can regulate the invasion and migration of oral cancer cells by targeting the miR-107/CDK6 signaling axis.