摘要
目的通过比较二乙基亚硝胺（DEN）诱发大鼠肝细胞癌发生过程中不同阶段的蛋白质表达谱，筛选在大鼠肝癌前病变阶段起重要作用的蛋白质分子。方法大鼠分为DEN组和正常对照组，诱癌过程中定期处死动物并取其肝组织；以γ-谷氨酰转肽酶染色阳性的肝细胞增生灶为标志，识别和获取肝癌前病变的组织标本。用双向电泳及基质辅助激光解吸飞行时间串联质谱法对大鼠正常肝组织、癌前病变组织和肝癌组织的全蛋白质组表达谱进行比较和鉴定。用Western blot和RT—PCR方法对部分差异表达蛋白质（如层黏连蛋白受体1和鲱精胺酶）进行表达水平的验证。结果筛选出表达水平差异≥2倍的蛋白质共82种，其中在癌前病变阶段即发生明显改变的差异表达蛋白质有47种，包括在正常对照组、癌前病变组、肝癌组呈现依次上调的层黏连蛋白受体1等8种蛋白质和在上述组织中呈现依次下调的鲱精胺酶等22种蛋白质。Western blot和RT—PCR方法对层黏连蛋白受体1和鲱精胺酶表达水平的验证结果与双向电泳的结果相似。结论大鼠肝癌形成过程中不同阶段的蛋白质表达谱有所不同；对癌前病变的差异表达蛋白质如层黏连蛋白受体1和鲱精胺酶等进行深入探讨，有望为人类肝癌的早期诊断和治疗提供线索。
Objective To screen the differentially expressed proteins especially at the precancerous stage of diethylnitrosamine （DEN） induced hepatocarcinogenesis by comparative proteome research. Methods Rats were divided into normal and DEN groups and sacrificed periodically. The liver samples were stained with γ-glutamyl transpeptidase （GGT） and HE to distinguish the preneoplastic lesion （pre-HCC） from the normal and HCC tissues. The two-dimensional electrophoresis （2-DE） and mass spectrometry （MALDI- TOF-MS/MS） were then applied to analyze the differentially expressed protein between pre-HCC and normal tissues, pre-HCC and HCC, as well as HCC and normal tissues. A few of the candidate proteins such as laminin receptor 1 （67LR） and agmatinase were validated by Western blot and RT-PCR. Results Totally,there were 82 proteins that differentially expressed two fold or more in one kind of tissues sample than the other, 47 of which occurred in the pre-HCC tissues. Eight proteins including 67LR were consistently up- regulated from normal tissue to pre-HCC and then to HCC tissues, while 22 proteins including agmatinase showed progressively down-regulated in these tissues samples. Conclusion The protein expression profiles are different during the process of hepatocarcinogenesis. Further study on the differentially expressed protein, especially these upregulated in the precancerous stage such as 67LR and agmatinase, might contribute to prevention and early diagnosis of human HCC.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2009年第9期669-674,共6页
Chinese Journal of Hepatology
基金
国家自然科学基金（30560167）
广西自然科学基金（桂科自0728195）
广西大型仪器测试基金（513-2007-092）
国家高科技研究发展计划（863计划）（2006AA02A308）
