天津医药

• 细胞与分子生物学 • 上一篇    下一篇

干扰LOX基因对喉癌Hep-2细胞增殖、侵袭及放疗敏感性的影响

董凯峰   

  1. 河北医科大学第一医院
  • 收稿日期:2013-09-09 修回日期:2014-01-09 出版日期:2014-05-15 发布日期:2014-05-15
  • 通讯作者: 董凯峰

The effect on proliferation, invasion and radiosensitivity of laryngeal cancer Hep-2 cells by interferencing LOX gene expression

  • Received:2013-09-09 Revised:2014-01-09 Published:2014-05-15 Online:2014-05-15

PDF (PC)

3007

摘要： 目的 探讨抑制赖氨酰氧化酶（LOX）基因表达对喉癌Hep-2细胞增殖、侵袭及放疗敏感性的影响。方法 转染LOX干扰质粒的 Hep-2细胞株，RT-PCR 检测 LOX mRNA,Western blot检测喉癌Hep-2细胞中LOX蛋白及细胞增殖侵袭相关蛋白Ki-67、PCNA、MMP-2/9的表达情况；MTT法检测不同剂量射线对Hep-2细胞生存率的影响，流式细胞术（FCM）检测 Hep-2 细胞凋亡。结果 转染 LOX干扰质粒后 Hep-2细胞中,LOX蛋白和 mRNA 的表达水平下调；Western blot结果显示转染组细胞LOX、Ki-67、PCNA、MMP-2/9蛋白表达明显低于阴性对照组和空白对照组；同时不同剂量射线对Hep-2细胞生存率影响呈剂量依赖性，转染组细胞转染后联合放疗细胞凋亡率明显高于空白及阴性对照组。结论 LOX干扰可特异下调LOX表达，抑制细胞增殖侵袭，作用机制可能与Ki-67、PCNA、MMP-2/9蛋白表达下调有关，并且可增强放疗敏感性，为临床提供一个新的靶点，提高喉鳞癌治疗效果。

关键词: 赖氨酰氧化酶, 增殖, 侵袭, 放疗

Abstract: Objective： to investigate the inhibiting lysyl oxidase (LOX) gene expression of laryngeal cancer cell-Hep-2 proliferation, invasion and influence of radiotherapy sensitivity.Methods: LOX interference plasmids were transfected into cells, The detection of LOX mRNA and protein expression were used by RT-PCR and Western blot., Ki-67,PCNA, MMP-2/9 protein expression were assied by western blot; flow cytometry (FCM) detection of apoptosis in Hep-2 cells. Results after LOX interference plasmid were transfected into Hep-2 cells, LOX protein and mRNA expression levels were cut down; Western blot results showed that LOX, Ki-67, PCNA, MMP-2/9 protein expressions were typically lower than the negative control group and control group; the apoptosis rate of transfection cells combined with radiotherapy was significantly higher than the control and the negative control group. Conclusions Downregulation of LOX expression can inhibit Hep-2 cells’ proliferation and invasion, enhanced radiosensitivity, the mechanism of this action may be related to the downregulation of Ki-67, PCNA, MMP-2/9 protein expression.In a word, this could provide a new target for clinical threapy, improving the effect of laryngeal squamous cell carcinoma.

Key words: lysyl oxidase, proliferation, invasion, radiation therapy