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Production of the Polyclonal Anti-human metallothionein 2a Antibody with Recombinant Protein Technology

Acta Biochimica et Biophysica Sinica 2006年第5期38卷 305-309页

作者： FaizM.M.T.MARIKAR The State Key Laboratory of Pharmaceutical Biotechnology Nanjing University Nanjing 210093 China

metallothionein 2a(MT2a)is a small stress response protein that can be induced by exposureto toxic *** is highly expressed in breast cancer *** this study,the cDNA encoding the humanMT2a protein was expressed as glutathione S-transferase(GST)fusion protein in Escherichia *** MT2a proteins were loaded onto 12% sodium dodecylsulfate-polyacrylamide gel and separatedby electrophoresis,the recombinant protein was visualized by Coomassie blue staining and the 33 kDarecombinant GST-MT2a fusion protein band was cut out from the *** gel slice was minced and used togenerate polyclonal *** of rabbit against MT2a protein allowed the production of hightiter polyclonal *** new polyclonal antibody recognized recombinant MT2a protein in Westernblot *** low-cost antibody will be useful for detection in various immuno-assays.

关键词： metallothionein 2a glutathione S-transferase polyclonal antibody

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Clinicopathological significance of altered metallothionein 2a expression in gastric cancer according to Lauren＇s classification

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Chinese Medical Journal 2013年第14期126卷 2681-2686页

作者： PAN Yuan-ming XING Rui CUI Jian-tao LI Wen-mei LU You-yong Department of Gastroenterology & Hepatology Chinese People's Liberation Army General Hospital Beijing 100853 China Laboratory of Molecular Oncology Key Laboratory of Carcino- genesis and Translational Research Ministry of Education Beijing Cancer Hospital/Institute School of Oncology Peking University Beijing 100142 China

Background Dysregulated metallothionein 2a （MT2a） has been implicated in carcinogenesis. The purpose of this study was to investigate the expression of MT2a in gastric cancer （GC） and its correlation with prognosis. Methods Reverse transcription-polymerase chain reaction and real-time polymerase chain reaction were used to detect the mRNA expression of MT2a in 12 GC cell lines, normal gastric epithelial GES-1 cells, and 36 GC and adjacent normal tissues. MT2a protein expression was determined in 258 GC tissues and 171 adjacent normal tissues by immunohistochemistry. Results MT2a mRNA expression was lower in GC cells and primary tumors than in GES-1 cells and adjacent normal tissues, respectively, High protein expression of MT2a was present in 130 of 171 normal tissues （76.0%） and in 56 of 258 GC tissues （21,7%; P 〈0.001）. MT2a protein expression was higher in well/moderately differentiated GC （22/54; 40.7%） than in poorly differentiated GC （34/204; 16.7%; P 〈0.001）. Moreover, the protein expression of MT2a was lower in diffuse-type GC （6/82; 7.3%） than in intestinal-type GC （50/176; 28.4%; P=0.0001）. Importantly, MT2a expression was an independent prognostic factor for GC, and decreased MT2a expression was associated with poor clinical outcome （P 〈0.001）. The expression status of MT2a could predict prognosis in intestinal and diffuse-type GCs. Conclusion Expression status of MT2a might be a useful prognostic biomarker for GC, especially when used in combination with Lauren＇s classification.

关键词： metallothionein 2a gastric cancer prognosis Lauren's classification

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