Kang Xiangdong, Zhang Long, Sun Jian, Ni Zhenhua, Ma Yanchun, Chen Xiaobo, Sheng Xia, Chen Teng
Central Laboratory, Putuo Hospital Affiliated Shanghai University of Traditional Chinese Medicine, Lanxi Road No. 164, Shanghai, 200062, China.
J Gastroenterol. 2008;43(8):618-25. doi: 10.1007/s00535-008-2208-3. Epub 2008 Aug 17.
Prohibitin (PHB) was found to be overexpressed in breast cancer and thus is suggested as a biomarker in that disease. A few studies have investigated the PHB expression pattern in gastric cancer by two-dimensional gel electrophoresis. Uncertainties still existed on whether PHB expression could indicate the differentiation and apoptosis degree of gastric cancer and whether PHB protein as well as anti-PHB antibody could be a biomarker in the serum of the gastric cancer patient. In this study, the expression levels of PHB protein and mRNA of the tissues as well as PHB antigen and anti-PHB antibody in serum of patients with gastric cancer were systemically examined.
Immunohistochemistry and real-time PCR were used to detect expression levels of PHB protein and mRNA in gastric cancer tissues. Recombinant PHB antigen was identified by Western blotting. The expression of PHB antigen and anti-PHB antibody was investigated by ELISA and TRFIA. Bcl-2 expression was examined by immunohistochemistry.
By immunohistochemistry and real-time PCR analyses, PHB protein and mRNA were both overexpressed in gastric cancer tissues compared to adjacent normal gastric tissues (P < 0.01). Moreover, an elevated PHB expression pattern paralleled the differentiation degree and Bcl-2 protein expression in gastric cancer. However, no significant differences of PHB protein and anti-PHB antibody expression were detected in serum of gastric cancer patients and that of healthy volunteers.
These results indicated that PHB could be a potential diagnostic and differentiation biomarker of gastric cancer for tissue-based detection by immunohistochemistry and real-time PCR, but not for serum-based detection.
研究发现禁蛋白(PHB)在乳腺癌中过表达,因此被认为是该疾病的一种生物标志物。一些研究通过二维凝胶电泳研究了PHB在胃癌中的表达模式。关于PHB表达是否能指示胃癌的分化和凋亡程度,以及PHB蛋白和抗PHB抗体是否可作为胃癌患者血清中的生物标志物,仍存在不确定性。在本研究中,系统检测了胃癌患者组织中PHB蛋白和mRNA的表达水平以及血清中PHB抗原和抗PHB抗体的表达情况。
采用免疫组织化学和实时定量PCR检测胃癌组织中PHB蛋白和mRNA的表达水平。通过蛋白质印迹法鉴定重组PHB抗原。采用酶联免疫吸附测定法(ELISA)和时间分辨荧光免疫分析法(TRFIA)研究PHB抗原和抗PHB抗体的表达。通过免疫组织化学检测Bcl-2的表达。
免疫组织化学和实时定量PCR分析显示,与相邻正常胃组织相比,胃癌组织中PHB蛋白和mRNA均过表达(P<0.01)。此外,胃癌中PHB表达升高的模式与分化程度及Bcl-2蛋白表达平行。然而,在胃癌患者血清和健康志愿者血清中未检测到PHB蛋白和抗PHB抗体表达的显著差异。
这些结果表明,PHB可能是一种潜在的胃癌诊断和分化生物标志物,可通过免疫组织化学和实时定量PCR进行基于组织的检测,但不适用于基于血清的检测。