干扰LOX基因对喉鳞癌Hep-2细胞增殖、侵袭及放疗敏感性的影响  

Effects of LOX Gene Expression on Proliferation, Invasion and Radiosensitivity of Laryngeal Cancer Hep-2 Cells

作　　者：董凯峰[1] 吕欣[1] 宋冬梅[1] 刘志明[1] 薛海涛[1] 张翠红[2] 

机构地区：[1]河北医科大学第一医院耳鼻咽喉头颈外科,050031 [2]白求恩国际和平医院放疗科

出　　处：《天津医药》2014年第5期417-420,共4页Tianjin Medical Journal

基　　金：河北省医学科学研究重点课题计划项目(项目编号:20120049)

摘　　要：目的探讨抑制赖氨酰氧化酶(LOX)基因表达对喉鳞癌Hep-2细胞增殖、侵袭及放疗敏感性的影响。方法将Hep-2细胞分为空白对照组(正常培养对照)、阴性对照组(转染试剂对照)、转染组(转染LOX siRNA),用Real-time PCR法、Western blot法分别检测各组细胞LOX、增殖细胞核抗原(Ki-67、PCNA)、基质金属蛋白酶(MMP)-2、MMP-9 mRNA和蛋白表达水平;MTT法检测不同剂量X射线(0、3、6、9、12、15、18 Gy)照射对Hep-2细胞生存率的影响,流式细胞术(FCM)检测Hep-2细胞凋亡情况。结果转染LOX siRNA后转染组Hep-2细胞中的LOX、Ki-67、PCNA、MMP-2、MMP-9 mRNA及蛋白表达明显低于阴性对照组和空白对照组。用剂量为12、15、18 Gy X射线照射Hep-2细胞后24 h,可明显抑制细胞生存率(P<0.05),并呈剂量依赖性。转染组联合放疗组的细胞凋亡指数明显高于空白对照组、阴性对照组及放疗组(%:79.11±1.26 vs 5.01±1.02、4.95±1.12、43.21±2.12,P<0.05)。结论转染LOXsiRNA可下调Hep-2细胞LOX表达,抑制细胞增殖,并且可增强放疗敏感性,作用机制可能与Hep-2细胞中Ki-67、PCNA、MMP-2、MMP-9表达下调有关。Objective To investigate the effect of lysyl oxidase （LOX） gene expression on proliferation, invasion and radiotherapy sensitivity in laryngeal cancer Hep-2 cells. Methods Hep-2 cells were divided into control group （normal cultured）, negative control group （transfection reagent） and transfection group （LOX siRNA transfected）. The expressions of LOX, Ki-67, PCNA, MMP-2 and MMP-9 mRNA were detected by real time-PCR and Western blot methods. The cell sur-vival rate and apoptosis irradiated by different doses of X-rays （0, 3, 6, 9, 12, 15 and 18 Gy） were detected by MTT and flow cytometry （FCM）. Results The expression levels of LOX, Ki-67, PCNA, MMP-2 and MMP-9 protein and mRNA were sig-nificantly lower in Hep-2 cells after transfection than those of control group and negative control group （P〈0.05）. The cell survival rate was significantly inhibited 24 hours after irradiation （12, 15 and 18 Gy） in a dose-dependent manner （P〈0.05）. The apoptotic rate of transfection cells combined with radiotherapy was significantly higher than that of control and the nega-tive control groups （%：79.11 ±177; 1.26 vs 5.01 ±177; 1.02, 4.95 ±177; 1.12, 43.21 ±177; 2.1,P〈0.05）. Conclusion The expression of LOX can be down-regulated after LOX siRNA transfection, inhibiting proliferation and enhancing radiosensitivity, which may be related to the down-regulation of Ki-67, PCNA, MMP-2 and MMP-9 protein expression.

关 键 词：喉肿瘤 蛋白赖氨酸6-氧化酶 RNA干扰 细胞增殖 放射疗法 赖氨酰氧化酶 

分 类 号：R739.65[医药卫生—肿瘤]