肝癌切除肝缺血再灌注损伤前后的蛋白质组学变化

Differential proteomic research of liver ischemia-reperfusion injury undergoing resection of hepatocellular carcinomas

背景:肝癌切除肝缺血再灌注损伤可促进微小转移灶的侵袭转移并引起线粒体损伤。目的:比较肝癌切除手术过程中肝正常组织缺血再灌注损伤前后蛋白质表达谱的改变,筛查发生肝缺血再灌注损伤应激的相关蛋白质分子标志物并探讨可能机制。方法:应用双向等电聚焦/聚丙烯酰胺凝胶电泳建立20例肝癌切除患者肝缺血再灌注前与灌注后15min肝正常组织的全细胞蛋白质和线粒体蛋白质表达图谱,并进行差异分析,应用基质辅助激光解吸离子化串联时间质谱对差异蛋白质点进行鉴定,并进行生物信息学分析。结果与结论:相同条件下对各组全细胞和线粒体蛋白样品分别进行3次双向电泳,筛选在3次电泳中均存在的差异点进行分析,在全细胞蛋白质图谱中共鉴定出5个差异蛋白,在线粒体蛋白质图谱中未鉴定出明显的差异蛋白。结果可见肝癌切除手术过程中肝缺血再灌注15min即可使细胞骨架蛋白、热休克蛋白等细胞保护蛋白表达发生改变,同时有促进肿瘤转移的风险,需要采取预防措施降低影响,但线粒体蛋白表达基本未发生改变。

BACKGROUND ： The ischemia reperfusion injury undergoes resection of hepatocellular carcinomas may promote the occult metastatic invasion and metastasis, and induce mitochondrial damage. OBJECTIVE： To reveal the protein expression changes of normal liver tissue after ischemia reperfusion injury undergoes resection of hepatocellular carcinomas, and to screen the hepatic ischemia reperfusion injury stress related protein and to explore the possible mechanism. METHODS： Normal liver tissue proteins and mitochondrial protein expression profiles were established in 20 liver cancer patients with hepatic ischemia reperfusion before and after perfusion for 15 minutes by isoelectric focusing/polyacrylamide gel electrophoresis, and the difference was analyzed. The matrix-assisted laser desorption inoization-time of flight mass spectrometry was used to identified different expression proteins, and the bioinformatic analysis was performed. RESULTS AND CONCLUSION： Under the same condition, two-dimensional electrophoresis of each whole cell and mitochondrial protein samples were performed for three times. Different proteins appeared in each pair profile were analyzed, tn the whole cell protein profiles, a total of 5 different proteins were identified and no significant difference of protein in mitochondrial profiles was identified. Liver ischemia reperfusion 15 minutes during hepatocellular carcinoma resection operation can change the expression of the cell protective protein such as cytoskeletal proteins and heat shock proteins, and can promote tumor metastasis risk and the preventive measures should be taken to reduce the impact. However, no changes happen in mitochondrial.