摘要
目的 :进一步从蛋白水平研究 SNC19编码蛋白与肿瘤转移的关系 ;方法 :免疫印迹法检测该蛋白在不同肿瘤细胞系及大肠癌组织中的表达 ;构建 Psectag2 A- SNC19(ORF)真核表达质粒 ,转染至乳腺癌细胞 BCAP37中瞬时表达 ,产物经亲和层析纯化 ,用 anti- His m Ab进行 Western blot鉴定其加工形式 ,并观察不同的加工产物是否具有明胶酶活性。结果 :免疫印迹法显示 ,在所选肿瘤细胞系和组织中 SNC19蛋白存在差异表达及翻译后加工 ;转染的 BCAP37细胞产生 12 0 k D和 75 k D两种形式的 SNC19蛋白 ,其中只有 75 k D的形式被证明具有明胶酶活性。结论 :SNC19蛋白在不同的肿瘤细胞系和肿瘤组织中存在差异表达及翻译后加工 ,其中 75 k D的形式被证明具有明胶酶活性。
Objective: To investigate the relationship between SNC19 protein and cancer metastasis. Methods: Expression of SNC19 protein in cancer cell lines and tissues was examined by Western blot analysis using anti SNC19 monoclonal antibody. In addition,Psectag2A SNC19(ORF) eukaryotic expression vector was constructed and transfected into BCAP37 cells.After the target protein was expressed and purified,processing forms of SNC19 protein were further identified using anti His mAb and each form was assayed for its gelatinase activity. Results: Different expression and post translational processing of the SNC19 proteins in the cancer cell lines and intestinal tissues were detected.BCAP37 cells transfected full length SNC19 (ORF) generated two different sized proteins in cell lysates,120 and 75 kD; 75 kD was detected to have proteolytic activity by gelatin zymography. Conclusion: SNC19 protein presents different expression and post translational processing in the cancer cells and tissues,of which 75 kD was identified to have gelatinase activity.
出处
《浙江大学学报（医学版）》
CAS
CSCD
2005年第1期38-42,共5页
Journal of Zhejiang University(Medical Sciences)
基金
国家 8 6 3高科技研究发展计划基金 (10 2 - 10 - 0 1-0 7)
浙江省自然科学基金 (ZB0 110 )项目 .
关键词
SNC19
蛋白质加工
转译后
真核表达
转染
明胶酶类
SNC19
Protein processing,post-translational
Eukaryotic expression
Transfection
Gelatinases
