华西口腔医学杂志 ›› 2019, Vol. 37 ›› Issue (4): 378-383.doi: 10.7518/hxkq.2019.04.007

• 基础研究 • 上一篇    下一篇

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

赵军方,查治安,谢卫红,王海斌,李新明,孙强(),孙明磊   

  1. 郑州大学第一附属医院口腔颌面外科,郑州 450052
  • 收稿日期:2018-12-21 修回日期:2019-02-19 出版日期:2019-08-01 发布日期:2019-08-23
  • 通讯作者: 孙强 E-mail:85244275@qq.com
  • 作者简介:赵军方,副主任医师,博士,E-mail:zhaolei197507@163.com
  • 基金资助:
    国家自然科学基金(81402231);河南省自然科学基金(182300410319)

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

Junfang Zhao,Zhian Zha,Weihong Xie,Haibin Wang,Xinming Li,Qiang Sun(),Minglei Sun   

  1. Dept. of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
  • Received:2018-12-21 Revised:2019-02-19 Online:2019-08-01 Published:2019-08-23
  • Contact: Qiang Sun E-mail:85244275@qq.com
  • Supported by:
    The National Natural Science Foundation of China(81402231);The National Natural Science Foundation of Henan Province(182300410319)

摘要:

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。

关键词: 口腔癌, 长链非编码RNA H19, 侵袭, 迁移

Abstract:

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 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.

Key words: oral cancer, long chain non-coding RNA H19, invasion, migration

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