用镭射激光捕获技术研究LEC鼠P53基因突变和肝癌发生的关系  

Use of laser capture microdissection (LCM) to study the relationship between P53 mutation of LEC rats and hepatocarcinogenesis

作　　者：程大也[1] 梁彬[1] 解芳[2] 梁红[3] 李丰[1] 

机构地区：[1]中国医科大学细胞生物学实验室 [2]中国医科大学高等职业技术学院检验教研室,辽宁沈阳110001 [3]中国医科大学肿瘤研究所,辽宁沈阳110001

出　　处：《中国现代医学杂志》2007年第12期1434-1437,共4页China Journal of Modern Medicine

基　　金：高等学校博士学科点专项科研基因(20050159023)

摘　　要：目的研究P53基因突变与原发性肝癌发生发展的关系。方法利用LEC鼠(long evan cinnamon)作为肝癌的动物模型。选取15周,37周,101周的LEC鼠15,14和10只,正常对照SD大鼠15只,取出肝脏组织进行切片。切片一部份用于GST-P免疫组化(immunohistochemistry,IHC)染色,另一部份利用镭射激光捕获技术(laser capture microdissection,LCM)定位切割,进行P53基因外显子5,6/7,8突变的分析。结果37周龄LEC鼠和101周龄LEC鼠GST-P阳性率高于对照组(P<0.005),15周龄LEC鼠GST-P阳性率与对照组相比差异无统计学意义(P>0.05)。P53基因在GST-P表达阳性肝炎组有3例突变(3/11),GST-P表达阳性肝癌组有10例发生突变(10/15),肝癌组显著高于肝炎组(P<0.05)。结论P53的突变发生在肝癌形成的早期阶段,随着发展过程,突变率明显增高。GST-P虽然特异性不强,但是不失为一种灵敏的判断肝损伤的指标。[Objective] To investigate the relationship of P53 mutation and primary hepatocareinogenesis. [Methods] Use the LEC rats as animal model, choose 15 15 weeks LEC rats, 14 37 weeks LEC rats and 10 101 weeks LEG rats as experiment group, 15 SD rats as control, took out liver and sectioned. Some sections were used for immunohistochemieal staining, the others were used for analysis of P53 mutation by LCM. [Results] Compared with the controls, GST-P expression ratios between 37 weeks and 101 weeks LEC rats without 15 weeks LEC rats were all higher than the controls （P 〈0.005）. Ratio of P53 mutation in hepatoma group with GST-P positive （10/15） was higher than hepatitis group （3/11） （P 〈0.05）. [Conclusion] P53 mutation ran through the initiative and late stages of hepato-carcinogenesis. With the progression, mutation ratio increases. Although GST-P specificity is not better, GST-P is a sensitive marker to define hepatic injury.

关 键 词：P53 镭射激光捕获技术 LEC鼠 突变 

分 类 号：R735.7[医药卫生—肿瘤]