Ras及Rab作用因子1基因在肝癌中的表达

Expression of Ras and Rab interactor 1 in liver cancer

目的通过检测位于大鼠肝癌模型的热点数量性状位点（QTL）区域内Ras及Rab作用因子1（RIN1）mRNA的表达，初步探讨RIN1与肝癌的关系。方法17只近交系雄性F344大鼠，12只喂食含0．06％3＇-甲基．4-二甲氨基偶氮苯（3＇-Me。DAB）的饲料以诱导肝癌模型（诱癌组），其余5只则喂食普通饲料（对照组）。20周后取各大鼠肝脏进行组织病理学检测以确认肝癌模型是否制备成功。用定量反转录PCR（qRT．PCR）分析大鼠肝癌组织、人肝癌组织（标本来自2010年1月至2011年12月的21例肝癌患者，其中11例是中山大学肿瘤医院、5例为本院行手术切除的原发性肝癌患者，另有5例患者标本购自上海芯超生物科技有限公司）及人肝癌细胞系（Hep3B、Huh7和HepG2）中RIN1mRNA的表达，并分别以正常对照F344大鼠肝脏、人肝癌癌旁组织及人正常肝细胞系（HL7702）作为相应对照进行对比。按年龄、肿瘤大小、病理组织分级、甲胎蛋白（AFP）和乙肝表面抗原（HBsAg）对21例肝癌患者进行分组，对比各亚组间RIN1mRNA表达差异，分析RIN1mRNA表达与肝癌患者临床指标的关系。结果诱癌组各大鼠肝脏明显肿大，形态失常，肉眼可见多个瘤体；组织学检测显示诱癌组F344大鼠肝脏存在肝细胞癌，肝癌模型制备成功。诱癌组大鼠肝癌组织RIN1mRNA表达量明显高于对照组（P〈0．05），为对照组大鼠肝脏的172．44倍。HepG2、Hep3B和Huh7细胞RIN1mRNA表达量分别为HL7702细胞的2．49、2．06和1．16倍，差异没有统计学意义。在21个肝癌患者标本中，有14个（66．67％）癌组织的RIN1表达量较相应癌旁组织降低，其倍数为0．02～O．92；与癌旁组织相比，癌组织RIN1表达量整体中位倍数为O．44，显著低于癌旁组织（P=O．049）。肝癌组织RIN1的表达量与患者年龄、肿瘤大小、病理组织分级、AFP和HBsAg均无关（均P〉0．05）。结论RIN1mRNA在肝癌组织中表达异常，可能与肝癌发生相关。

Objective To investigate the relationship of Ras and Rab interactor 1 （RIN1） with liver cancer by analyzing the mRNA expression in the hotspot quantitative trait locus （QTL） region in rat models with liver cancer. Methods Seventeen inbred male F344 rats were either fed with foods containing 0.06% 3＇ - methyl- 4 - dimethylaminoazobenzene （3＇ - Me - DAB） （carcinogenesis group, n=12） or common food （control group, n=5）. This was followed by hepatic biopsy at week 20 to ensure that the liver cancer model was successfully constructed. Quantitative reverse transcript!on-polymerase chain reaction （qRT-PCR） was employed to assay the RIN1 mRNA expression in rat liver cancer tissues, human liver cancer tissues [21 liver cancer patients from Affiliated Cancer Hospital of Sun Yat-sen University （n=ll） , Guangzhou First People＇s Hospital, Guangzhou Medical University （n=5） and Shanghai Xinchao Biotech Co. Ltd. （n=5） during January 2010 and December 2011 ] and human liver cancer cell lines （Hep3B, Huh7 and HepG2）,for which the normal liver tissues of F344 rats, human pericancerous liver tissues and human HL7702 cell line served as controls. The liver cancer patients were divided into subgroups according to age, tumor size, histological grade, alpha- fetal protein （AFP） and HBsAg for comparison on RIN1 expression among subgroups and the correlation with clinical indices. Results Carcinogenesis group evidenced significant hepatomegaly characterized by aberrant appearance and multiple tumors on gross anatomy. The histological examination further confirmed the presence of hepatocellular carcinoma in F344 subgroup, liver cancer models were successfully. Carcinogenesis group yielded the RIN 1 mRNA expression 172.44 times higher than that in control group （P〈0.05）. Compared with HL7702 cells, the HepG2, Hep3B and Huh7 cell lines had a 2.49-, 2.06- and 1.16-times higher RIN1 expression （all P〉0.05）, respectively. Of 21 human liver cancer tissues, 14 （66.67%） had decreased RIN1 expression （range： 0.02-0.92 times, median： 0.44 times, P= 0.049） compared with pericancerous tissues. RIN 1 expression was not significantly associated with the age, tumor size, pathologic grading, AFP and HBsAg （all P〉0.05）. Conclusion The aberrant RIN1 mRNA expression in liver cancer tissues may be associated with the carcinogenesis of liver cancer.