Coronin-1 C蛋白表达与肝癌侵袭和转移相关

Increased coronin-1C expression is related to hepatocellular carcinoma invasion and metastasis

目的 采用膜蛋白质组学技术筛选与原发性肝癌侵袭和转移相关的分子,并加以验证.方法 以不同转移潜能的人肝癌细胞HCCLM9和MHCC97L为研究对象,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和电喷雾串联质谱技术比较和鉴定差异表达的膜蛋白,进一步行Westernblot、动物模型标本和临床标本验证. 结果电喷雾串联质谱鉴定表达差异条带中蛋白质14个,其中Western blot验证coronin-1 C在人肝癌细胞HCCLM9中灰度值coronin-1 C/整合素α3比值为7.31±0.73,在MHCC97L中为2.84±0.99.coronin-1C在HCCLM9细胞中的表达水平显著性升高（t=6.27,P〈0.05）.在裸鼠肝癌组织中免疫组织化学提示coronin-1C表达也明显升高.115例临床病理标本免疫组织化学显示,coronin 1C表达越强,肿瘤的侵袭程度越强、临床病期越晚.结论 Coronin-1C与肝癌的侵袭和转移相关.

Objective To search for hepatocellular carcinoma （HCC） invasion related biomarkers using the cell membrane proteomics approaches, and to validate the markers using experimental and clinical specimens. Methods The HCCLM9 and MHCC97L cells with a similar genetic background and remarkably different metastasis behaviors were used for comparative membrane proteome profiling using sodium dodecyl sulfate polyacrylamide gel electrophoresis and electrospray ionization mass spectrometry technologies. Candidate protein makers were further validated by western blot on cells, immunohistochemistry （IHC） on animal tumor tissues, and tissue micro-array on clinical specimens. Results The membrane proteins of MHCC97L and HCCLM9 cells were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis analyses.14 proteins were identified by ESI-MS/MS among the differential bands. Coronin-1C was overexpressed in HCCLM9 （7.31±0.73） versus MHCC97L （2.84±0.99） validated by western blot. Elevated coronin-1C expression was observed in liver cancer tissues of HCCLM9 nude mice. IHC study in 115 human HCC specimens demonstrated that patients with higher coronin-1C expession had more advanced stage. Conclusion The study suggests that coronin-1C could be a potential molecule to predict HCC invasive behavior.