中国实用口腔科杂志

中国实用口腔科杂志

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has-miR-16在舌鳞癌中的生物学功能及分子机制研究

李琦a，耿亚军b，郭菁a，鄂丽华a，徐溢涵a，金武龙a    

  1. 内蒙古医科大学附属医院 a 口腔科，b 肝胆外科，内蒙古 呼和浩特  010059
  • 出版日期:2017-06-15 发布日期:2017-07-11
  • 基金资助:

    内蒙古医科大学科技百万工程项目（2014YKDKJBW01）；内蒙古医科大学人才团队项目：口腔再生医学团队（NYTD-2015011）

  • Online:2017-06-15 Published:2017-07-11

摘要：

目的    探究has-miR-16在舌鳞癌发生发展中的作用及分子机制。方法    通过实时荧光定量PCR检测36例舌鳞癌与癌旁组织样本中has-miR-16的表达特征，并检测其在舌鳞癌细胞株CAL-27、SCC-9及正常人口腔黏膜成纤维细胞中的表达情况。转染has-miR-16 模拟物（mimics）至CAL-27和SCC-9细胞，采用噻唑蓝（MTT）比色法和创伤愈合试验检测其对细胞增殖及迁移的影响。通过生物信息学方法预测has-miR-16的靶基因，并采用双荧光素酶报告基因试验进行验证。结果    Has-miR-16在舌鳞癌组织的表达量明显低于癌旁组织（P < 0.05），其在CAL-27及SCC-9细胞中的表达量亦低于正常人口腔黏膜成纤维细胞（P < 0.05）。体外功能实验发现，过表达has-miR-16可显著抑制CAL-27与SCC-9细胞的增殖及迁移（P < 0.05）。生物信息学分析及双荧光素酶报告基因试验发现，成髓细胞瘤癌基因（myeloblastosis oncogene，MYB）是has-miR-16的靶基因。结论    has-miR-16在舌鳞癌组织及细胞系中低表达，过表达has-miR-16可抑制舌鳞癌细胞的增殖及迁移。has-miR-16在舌鳞癌中发挥抑癌作用，其作用机制可能通过调控MYB而实现。

关键词:  , has-miR-16, 舌鳞癌, 抑癌作用, 成髓细胞瘤癌基因

Abstract:

Objective    To study the role and molecular mechanism of has-miR-16 in the development of tongue squamous cell carcinoma （TSCC）. Methods    The expression of has-miR-16 was detected in 36 cases of TSCC tissues and matched normal tissues by using Quantitative real time PCR. The expression of has-miR-16 in TSCC cells （CAL-27 and SCC-9） and the normal human oral mucosa fibroblast cells （hOMF） was also analyzed by Quantitative real time PCR. The cell proliferation and migration of CAL-27 and SCC-9 were analyzed by MTT and wound healing assays after being transfected with has-miR-16 mimics or mimics control. The target gene of has-miR-16 was predicted by bioinformatics and verified by dual luciferase reporter assay. Results    The expression of has-miR-16 in TSCC tissues was significantly lower than matched normal tissues （P < 0.05）. The expression of has-miR-16 was also down-regulated in CAL-27 and SCC-9 cells compared with hOMF cells （P < 0.05）. Over-expression of has-miR-16 inhibited cell proliferation and migration in CAL-27 and SCC-9 cells （P < 0.05）. MYB （Myeloblastosis oncogene） was the target gene of has-miR-16 in TSCC. Conclusion    Has-miR-16 is down-regulated in TSCC tissue and cell lines；over-expression of has-miR-16 inhibites CAL-27 and SCC-9 cells proliferation and migration；has-miR-16 acts as tumor suppressor in TSCC，which may play its role via regulating MYB.

Key words: has-miR-16；tongue squamous cell carcinoma, TSCC；suppressor；myeloblastosis oncogene, MYB