CAJ | 학술논문

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.
목적 채용막단백질조학기술사선여원발성간암침습화전이상관적분자,병가이험증.방법 이불동전이잠능적인간암세포HCCLM9화MHCC97L위연구대상,채용십이완기류산납-취병희선알응효전영화전분무천련질보기술비교화감정차이표체적막단백,진일보행Westernblot、동물모형표본화림상표본험증. 결과전분무천련질보감정표체차이조대중단백질14개,기중Western blot험증coronin-1 C재인간암세포HCCLM9중회도치coronin-1 C/정합소α3비치위7.31±0.73,재MHCC97L중위2.84±0.99.coronin-1C재HCCLM9세포중적표체수평현저성승고(t=6.27,P<0.05).재라서간암조직중면역조직화학제시coronin-1C표체야명현승고.115례림상병리표본면역조직화학현시,coronin 1C표체월강,종류적침습정도월강、림상병기월만.결론 Coronin-1C여간암적침습화전이상관.
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.