3'-Me-DAB诱导大鼠肝癌发生过程中p53及相关基因mRNA的定量分析  

Quantitative Analysis of p53and Rel ated Genes mRNA in Rat Hepatocarcinogenesis Induced by 3' -Me-DAB

作　　者：付艳[1] 邓伟国[2] 李玉林[3] 杉山俊博[4] 

机构地区：[1]吉林大学第一医院妇产科,吉林长春130021 [2]吉林大学公共卫生学院营养与食品卫生教研室,吉林长春130021 [3]吉林大学基础医学院病理教研室,吉林长春130021 [4]秋田大学医学部生化学第一讲座

出　　处：《癌症》2003年第1期35-41,共7页Chinese Journal of Cancer

基　　金：日本癌研究促进基金资助

摘　　要：背景与目的:有关肝癌中p53基因突变及p53蛋白表达异常已有报道,但其在mRNA水平上的变化尚不清楚。为了解在肝癌发生、发展和预后过程中p53、谷胱甘肽转硫酶P(glutathioneS-transferaseP,GST-P)、甲胎蛋白(α-fetoprotein,AFP)和白蛋白mRNA水平的变化,本研究定量分析癌前病变和癌灶中GST-P、AFP和白蛋白mRNA的量。方法:在3'-甲基-4-二甲胺偶氮苯(3'-methyl-4-dimethylaminoazobenzene,3'-Me-DAB)诱发F344大鼠肝癌过程中,用激光捕获显微取样仪(LCM)准确获得大鼠肝脏中微小癌灶或癌前病变组织后,采用LightCyclerTMV3System实时(real-time)RT-PCR定量分析这些病灶组织中mRNA水平。结果:在实验的第6、12和24周,癌前病变组织中p53mRNA量均显著高于正常组织(P≤0.001)。从第6周到第24周,癌前病变组织中p53mRNA逐渐降低(P<0.01)。癌组织中p53mRNA含量高于正常组织,低于同期癌前病变组织(P=0.028和0.0136)。第24周的癌前病变组织或癌组织中细胞核呈p53强染色。各实验期,癌前病变中GST-PmRNA量明显高于正常组织和癌组织(P<0.001)。癌组织中AFPmRNA的表达量显著高于癌前病变组织和正常组织(P<0.001),白蛋白mRNA的表达量显著低于这两种组织(P<0.01)。GST-P和AFP分别在癌前病变组织和癌组织中呈灶状强表达。结论:GST-BACKGROUND &OBJECTIVE:p53g ene mutations and abnormal expression of p53in hepato carcinoma have been reported,but alteration in mRNA level is not yet understood.In order to find out the alt eration in mRNA levels of p53,g lutathione S-transferase P (GST-P),alpha-fetoprotein(AFP),and albumin in g enesis ,developing,and prog nosis of hepatocarcinoma ,quantitative analysis of mRNA levels of p 53,GST-P,AFP,and albumin in prehepatocarcin oma and hepatocarcinoma foci was performed.METHODS :During hepatocarcinog enesis of F344rat induced by 3' -methyl-4-dimethylamino-azobenz ene(3'-Me-DAB),the levels of these mRNAs were quantitatively analyzed by Lig htCycler TM V3System real-time RT-PCR after capturing accurately micro-foci in prehepatocarcinoma and hepato-car cinoma of rats with laser capture microdissec tion(LCM).RESULTS :At the 6 th ,12 th and24 th experiment weeks,the p53mRNA level s in all of prehepatocarcinoma foci were markedly hig her than those in th e adjacent normal tissues (all of P≤0.001),and g radually decreased from the 6 th week to the 24 th week(P<0.01).The content of p53mRNA in hepatocarcino ma foci was hig her than that in normal tissue(P=0.028and 0.013),but lower than that in prehepatocar cinoma foci.At 24 th weeks ,the sections of livers exhibi ted intensive immunostaining of p53protein in prehepatocarcinoma and h epatocarcinoma foci.At any time-po int of experiment,GST-P mRNA levels in pre hepatocarcinoma foci were sig nific antly hig her than those in the adjacent nor mal liver tissue and hepatocarcinom a foci (all of P<0.001).The concentration of AFP mRNA was th e hig hest (P<0.001)and that of albumin mRNA was the lowes t(P<0.01)in hepatocarcinoma foci as compared with adjacent normal tis sue and prehepatocarcinoma foci.Th e GST-P protein and AFP protein were expressed strong ly in prehepatocarci noma and hepatocarcinoma foci,respectively.CONCLUSION:GST-P and AFP mRNA overexpressed in prehepatocarcinoma and hepatocarcinoma foci respect iv

关 键 词：3′-Me-DAB 诱导 大鼠 MRNA P53基因 RT-PCR 免疫组织化学 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]