目的 筛查肝癌肝移植术后转移复发关键分子。方法 取具有相同疾病背景肝癌肝移植患者原发瘤标本,根据临床随访资料,分为复发和未复发组(各取6例)。激光显微切割分别从原发瘤标本肿瘤组织中切取同质肝肿瘤细胞,提取总蛋白,按组别等量混合蛋白后,分别进行稳定同位素轻重链标记,2D—LC—MS/MS鉴定差异蛋白;免疫组织化学及免疫印迹验证选取的差异蛋白。结果 质谱鉴定出149个有定量信息蛋白,其中表达上调2倍以上蛋白29个,表达下调2倍以上蛋白共23个。利用免疫组织化学和蛋白免疫印迹对其中上调差异蛋白Capn4进行了验证,验证结果和质谱鉴定的结果相吻合。结论 肝癌肝移植术后转移复发与多种蛋白表达改变相关,其中Capn4高表达可能在复发转移中发挥作用,可能作为肝癌肝移植术后转移复发潜在的分子靶标。
Objective To screen the key molecule associated with metastasis and recurrence of hepatocellular carcinoma (HCC) after liver transplantation. Methods The primary liver cancer samples of the patients with HCC after liver transplantation who had the same disease background were divided into the recurrent group and non-recurrent group (6 cases in each group) according to the follow-up data. The homogeneous HCC cells were obtained from HCC tissues, and the total protein was extracted,labeled with the light and heavy cICAT reagents respectively, and identified with two dimensional liquid chromatography coupled with tandem mass spectrometry. Immunohistochemistry and Western blotting were performed to examine the differential expression of the candidate proteins. Results Total 149 proteins including 29 upregulated 2-folds and 23 down-regulated 2-folds proteins were identified, of these, Capn4 was found to be uniquely over-expressed in the recurrent group when compared to the non-recurrent group. Immunohistochemistry and Western blotting confirmed this difference. Conclusion The metastasis and recurrence of HCC after liver transplantation may be correlated with the special variation of the protein expression profile. The overexpression of capn4 may play a crucial role in that process, and Capn4 may serve as a potential molecular target associated with the metaststic HCCs after liver transplantation.