18srRNA作为内参在肺癌相关基因LSCC-3研究中的应用  

18srRNA ACTING AS AN INTERNAL STANDARD OF SEMI-QUANTITATIVE RT-PCR IN THE STUDY OF LSCC-3,A LUNG CANCER RELATED GENE

作　　者：陈应泰[1] 姜冠潮[1] 王丹蕾[1] 王俊[1] 

机构地区：[1]北京大学人民医院胸外科,胸部微创中心,北京100044

出　　处：《广西医科大学学报》2005年第2期209-212,共4页Journal of Guangxi Medical University

摘　　要：目的:利用以18srRNA为内参照系统的半定量RT-PCR方法对肺癌相关基因LSCC-3在肺癌组织和正常肺组织中的差异表达进行研究并探讨18srRNA作为内参照系统的可行性及其系统优化。方法:分别提取肺癌组织和癌旁正常肺组织共计50对标本的总RNA,根据人肺癌相关基因LSCC-3和人18srRNA序列分别设计引物,以18srRNA作为内参照物进行双扩增RT-PCR反应,对反应体系及过程进行优化并分别进行三次反应,三次反应产物经琼脂糖凝胶电泳后以凝胶成像分析系统行半定量分析,并对结果进行统计学处理。结果:经优化反应系统,以18srRNA作为内参照物的半定量RT-PCR反应结果稳定、三次反应结果经t检验显示差异无统计学意义(P>0.05);经配对t检验,人肺癌相关基因LSCC-3在肺癌组织中表达上调,与癌旁正常组织间存在明显的差异表达(P<0.005),与同期进行的Northernblot实验结果完全符合。结论:由于18srRNA在各种不同功能状态的不同细胞内的量的相对稳定,经过优化条件,以18srRNA作为内参照物的半定量RT-PCR系统的结果是稳定、可信的。Objective:To study the differential expression of Gene LSCC-3 in the lung cancer and normal tissues nearby; and to estimate the feasibility and advantages of 18srRNA as an internal standard in the semi-quantitative RT-PCR.Methods:Collecting RNA of 50 pairs of lung cancer and normal tissues nearby; designing the special Primers for LSCC-3 and 18srRNA;performing Semi-quantitative RT-PCR, which used 18srRNA as the internal standard of the lung cancer and normal tissues nearby for three times;and analyzing the results gotten form the semi-quantitative RT-PCR.Result:the results of the three semi-quantitative RT-PCRs were similar and there was no obvious difference among the results (P>0.05); the expression of Gene LSCC-3 in the lung cancer and normal tissues nearby was obviously differential (P<0.005) which accorded to the result of northern blot performed at the same time.Conclusion:Because of the stabilization of expression of 18srRNA in the distinct cells, after an optimization of the conditions of the trial, the results of a semi-quantitative RT-PCR, used 18srRNA as an internal standard, are stable and believable.

关 键 词：肺癌 肺癌相关基因 18srRNA 半定量RT—PCR 

分 类 号：R734.2[医药卫生—肿瘤]