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, miR-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, miR-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 co-transfection 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 coregulated by H19 and miR-107 (P<0.05).
Conclusions: 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.
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）。siRNA H19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，anti-miR-107共转染可促进siRNA H19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 lncRNA H19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。.
Keywords: invasion; long chain non-coding RNA H19; migration; oral cancer.