Song Hai-Yan, Liu Yin-Kun, Feng Ju-Tao, Cui Jie-Feng, Dai Zhi, Zhang Li-Jun, Feng Jiu-Xian, Shen Hua-Li, Tang Zhao-You
Liver Cancer Institute, Zhongshan Hospital, Fudan University, 136 Yi Xue Yuan Road, 200032 Shanghai, People's Republic of China.
J Cancer Res Clin Oncol. 2006 Feb;132(2):92-8. doi: 10.1007/s00432-005-0044-x. Epub 2005 Nov 1.
A comparative proteomic approach was used to identify and analyze proteins related to metastasis of hepatocellular carcinoma (HCC).
Proteins extracted from 12 HCC tissue specimens (six with metastases and six without) were separated by two-dimensional gel electrophoresis (2-DE). The protein spots exhibiting statistical alternations between the two groups through computerized image analysis were then identified by mass spectrometry. In addition immunohistochemistry (IHC), Western blotting and RT-PCR were performed to verify the expression of certain candidate proteins.
16 proteins including HSP27, S100A11, CK18 were annotated by mass spectrometry, relevant to chaperone function, cell mobility, cytoskeletal architecture, respectively. Most were previously unconnected with metastasis of HCC. Of these HSP27 was found overexpressed consistently in 2-DE patterns of all metastatic HCC tissues compared with nonmetastatic ones. IHC and Western blotting of HCC tissues confirmed this difference while RT-PCR did not.
There are various proteins joined together in HCC metastasis. The overexpression of HSP27 may serve as a biomarker for early detection and therapeutic targets unique to the metastatic phenotype of HCC.
采用比较蛋白质组学方法鉴定和分析与肝细胞癌(HCC)转移相关的蛋白质。
从12例HCC组织标本(6例有转移,6例无转移)中提取的蛋白质通过二维凝胶电泳(2-DE)进行分离。然后通过计算机图像分析在两组之间表现出统计学差异的蛋白质斑点通过质谱进行鉴定。此外,进行免疫组织化学(IHC)、蛋白质印迹法和逆转录-聚合酶链反应(RT-PCR)以验证某些候选蛋白质的表达。
通过质谱鉴定出16种蛋白质,包括热休克蛋白27(HSP27)、钙结合蛋白A11(S100A11)、细胞角蛋白18(CK18),分别与伴侣功能、细胞迁移、细胞骨架结构相关。大多数以前与HCC转移无关。其中,与非转移性HCC组织相比,HSP27在所有转移性HCC组织的2-DE图谱中均持续过表达。HCC组织的免疫组织化学和蛋白质印迹法证实了这种差异,而逆转录-聚合酶链反应未证实。
在HCC转移中有多种蛋白质共同起作用。HSP27的过表达可能作为HCC转移表型的早期检测生物标志物和独特的治疗靶点。