MicroRNA-196a 在喉癌组织中的表达及其 对喉癌细胞侵袭和转移的影响

Expression of miR-196a in laryngeal carcinoma and its effect on invasion and metastasis of laryngeal carcinoma cells

摘要:

目的 探讨microRNA-196a（miR-196a）在喉癌组织中表达及其对喉癌细胞侵袭和转移的影响。 方法 选取2015 年1 月—2017 年12 月金华市中心医院70 例喉癌手术切除标本的喉癌组织和癌旁组织。将 人喉癌Hep-2 细胞分为空白对照组、阴性对照组及siRNA 组。siRNA 组细胞转染pcDNA/miR-196a，阴性 对照组细胞转染pcDNA/miR-NC，空白对照组不做处理。采用逆转录聚合酶链反应（RT-PCR）测定癌旁 组织和喉癌细胞中miR-196a 水平；Transwell 小室测定喉癌细胞侵袭和迁移能力；Western blotting 检测喉癌 细胞磷脂酰肌醇-3- 激酶（PI3K）、磷酸化蛋白激酶B（p-PKB）、Bax 及Bcl-2 蛋白水平；RT-PCR 测定喉 癌细胞PI3K、p-PKB、Bax 及Bcl-2 mRNA 水平。结果 喉癌组织中miR-196a 水平高于癌旁组织（P <0.05）。 与空白对照组和阴性对照组比较，siRNA 组喉癌细胞中miR-196a 水平降低（P <0.05），细胞侵袭和迁移能 力下降（P <0.05），PI3K、p-PKB 及Bcl-2 蛋白和mRNA 水平降低（P <0.05），Bax 蛋白和mRNA 水平升 高（P <0.05）；空白对照组与阴性对照组细胞各指标比较，差异无统计学意义（P >0.05）。结论 喉癌组织中 miR-196a 水平升高，沉默miR-196a 可降低喉癌细胞的侵袭和迁移能力，其机制可能与PI3K/PKB 信号通路 有关。

Abstract:

Objective To investigate the expression of miR-196a in laryngeal carcinoma, the effect of miR- 196a on invasion and metastasis of laryngeal carcinoma cells and its mechanism. Methods A total of 70 cases of laryngeal cancer tissues and its adjacent tissues from laryngeal cancer which were surgically removed were collected from the department of otorhinolaryngology in Jinhua Central Hospital from January 2015 to December 2017. Human laryngeal carcinoma Hep-2 cells were divided into blank control group, negative control group and siRNA group. The siRNA group cells were transfected with pcDNA/miR-196a and the negative control group cells were transfected with pcDNA/miR-NC, while the blank control group cells were not treated. The miR-196a levels in tissues and laryngeal carcinoma cells were determined by reverse transcription-polymerase chain reaction (RT-PCR). The transwell chamber was used to measure the invasion and migration ability of laryngeal cancer cells. Western blot was used to detect the levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-PKB), Bax and Bcl-2 in laryngeal carcinoma cells. The mRNA levels of PI3K, p-PKB, Bax and Bcl-2 in laryngeal carcinoma cells were determined by RT-PCR. Results The level of miR-196a in laryngeal carcinoma tissues was higher than that in adjacent tissues (P < 0.05). Compared with the blank control group and the negative control group, the level of miR-196a in the siRNA group was decreased (P < 0.05), the cell invasion and migration ability were decreased (P < 0.05), the levels of PI3K, p-PKB, Bcl-2 protein and mRNA were decreased (P < 0.05), and the levels of Bax protein and mRNA were increased (P < 0.05).There were no significant difference in each index between the blank control group and the negative control group (P > 0.05). Conclusions The level of miR-196a is elevated in laryngeal carcinoma. Silence of miR-196a can reduce the invasion and migration of laryngeal carcinoma cells, and the mechanism may be related to PI3K/PKB signaling pathway.