沉默赖氨酰氧化酶基因对喉癌Hep-2细胞凋亡及化疗敏感性的影响

Effects on apoptosis and chemosensitivity in human laryngeal cancer Hep-2 cells by silencing the lysyl oxidase gene expression

目的探讨沉默赖氨酰氧化酶(LOX)基因表达对喉鳞癌Hep-2细胞凋亡及化疗敏感性的影响。方法将LOX siRNA转染至Hep-2细胞,免疫细胞化学及免疫印迹(Western blot)检测喉鳞癌Hep-2细胞中LOX蛋白表达情况,实时聚合酶反应(RT-PCR)检测LOX mRNA;MTT法检测不同浓度顺铂对Hep-2细胞生长抑制率的影响;流式细胞术(FCM)检测Hep-2细胞凋亡。结果转染LOX siRNA后Hep-2细胞中LOX蛋白和mRNA表达水平下调(P<0.05);用浓度为2、3、4 mg/L顺铂作用Hep-2细胞后,可明显抑制细胞生长(P<0.05)。转染组细胞联合顺铂作用后细胞凋亡指数及生长抑制率显著高于阴性对照组、空白对照组、顺铂组及转染组(均P<0.05)。结论特异性抑制LOX表达可诱导Hep-2细胞凋亡,并增强喉鳞癌Hep-2细胞对顺铂的敏感性。

【Objective】 To investigate the effects of silencing the lysyl oxidase（LOX） gene expression in laryngeal cancer Hep-2 cells on apoptosis and chemosensitivity. 【Methods】 LOX siRNA were transfected into Hep-2 cells, LOX mRNA/protein were detected by Real-time PCR, Western blotting and immunocytochemistry assay; The proliferation was assessed by MTT, and the apoptosis of Hep-2 cells was detected by flow cytometry（FCM）. 【Results】 In the LOX siRNA transfected group, LOX mRNA and protein expression levels decreased; the apoptosis index was significantly higher than the control group and negative control group. MTT assay showed that cell proliferation of siRNA transfected group was significantly lower than the negative control group and control group. The inhibition rate of cell in siRNA transfected group administrated with cisplatin was significantly higher than the control group and negative control group. 【Conclusion】 The specific inhibition of LOX can induce apoptosis of Hep-2 cells and enhance the sensitivity of Hep-2 cells to chemotherapeutic drugs.