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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 key molecules of metastasis and recurrence of liver cancer after liver transplantation. Methods Tumor specimens of primary liver cancer patients with liver transplantation under the same disease background were collected and divided into recurrence and non-recurrence groups according to the clinical follow-up data (6 in each). Laser microdissection was taken from the primary tumor tumor tissue homogenate liver tumor cells, total protein was extracted, the same amount of protein were mixed by group, respectively, stable isotope light and heavy chain markers, 2D-LC-MS / MS identification of differences Proteins; Immunohistochemistry and Western blotting to validate selected proteins. Results A total of 149 proteins with quantitative information were identified by mass spectrometry, of which 29 were up-regulated more than 2-fold and 23 were down-regulated more than 2-fold. The protein Capn4 was up-regulated by immunohistochemistry and Western blotting. The validation results were in good agreement with those by mass spectrometry. Conclusion The recurrence and metastasis of hepatocellular carcinoma after liver transplantation is related to the changes of many protein expressions. The high expression of Capn4 may play a role in the recurrence and metastasis, which may be used as a potential molecular target of metastasis and recurrence of liver cancer after liver transplantation.