RNAi抑制NF-κB通路相关基因对喉鳞癌凋亡及转移的影响  

The affection of RNAi inhibiting NF-κB pathway related genes on apoptosis and invasion of laryngeal squamous cell carcinoma

作　　者：黄带发[1] 富伟能[2] 孙开来[2] 徐振明[1] 董卫东[1] 

机构地区：[1]沈阳军区总医院,辽宁沈阳110015 [2]中国医科大学基础医学院医学遗传学教研室,辽宁沈阳110001

出　　处：《现代肿瘤医学》2008年第3期351-354,共4页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:30171008);国家863计划子课题(2002BA711A08-18)

摘　　要：目的:用RNA干涉方法探讨NF-κB通路参与喉癌发生、发展的可能机制。方法:RT-PCR、Western Blot检测喉鳞癌组织中NF-κB通路中相关基因表达,RNA干涉方法抑制p65,p50表达,流式细胞仪,TUNEL方法检测细胞凋亡,Transwell检测细胞侵袭力。结果:36例喉鳞癌组织中21例p65mRNA表达上调(58.33%)高于癌旁组织(P=0.012),13例出现p50mRNA表达上调(36.11%),但与癌旁组织比较无显著差异(P=0.602)。喉鳞癌组织的p65蛋白表达明显较癌旁组织增强(P=0.044),而p50蛋白表达无显著差异。特异siRNA转染Hep2细胞明显抑制p50,p65基因表达,该作用可持续10天。RNAi抑制p65基因表达使Hep2细胞凋亡率增加,在转染后第7天最高达29.89%,与对照组比较升高超过10倍(P=0.020);同时使Hep2细胞侵袭力明显下降(P=0.003),而干涉p50基因对细胞凋亡及细胞侵袭性影响不明显。结论:RNAi抑制p65表达诱导Hep2细胞凋亡、抑制细胞侵袭力,提示抑制p65表达与常规治疗结合可能成为改善喉癌预后的选择。Objective：Using RNAi to explore the effects of NF -κB pathyway on laryngeal squamous cell carcinoma （LSCC）. Methods： Semi -quantitative RT -PCR and Western blotting was used to detect expression of NF -κB pathyway. RNA interfering （RNAi） was used here knockdown p65 and p50 genes, then cell apoptosis was measured by flow cytometer as well as fluorescence staining assay, and cell invasion was measured by Transwell assay. Results： p65mRNA up - regulated in 58.33% of LSCC（21/36） , it was significantly different compared with the paired adjacent normal laryngeal tissues （PANL） （P = 0. 012 ）, but p50 not significantly different compared with PANL（ P = 0. 602）. p65 protein enhanced in LSCCs compared with PANLs（ P = 0.044）. Transfection of siRNA resulted in remarkable suppression on expression of 1965 and p50 mRNA in Hep2 cells, this effect can maintain 10 days. Moreover, down- regulation of p65 mRNA enhanced Hep2 cell apoptostic rate, which reached up to 29.89% on the 7^th day, over 10-fold （P=0. 020） , as well as the invasion of Hep2 cells were inhibited dramatically （P = 0. 003 ）. However, it had less effect on Hep2 cell after down - regulation p50 mRNA. Conclusion ： Down - regulation of p65 mRNA sensitized Hep2 cells to apoptosis as well as restrained invasion, which might be a choice for ameliorating the prognosis of LSCC.

关 键 词：喉 鳞状细胞癌 NF—κB通路 RNA干涉 Hep2细胞 

分 类 号：R739.65[医药卫生—肿瘤]