人食管癌相关基因4蛋白的亚细胞定位、纯化和抑癌功能研究

Studies on the subcellular localization, purification and tumor-suppressing function of human e-sophageal cancer related gene 4 protein in esophageal carcinoma

目的检测人食管癌相关基因4（ECRG4）蛋白的亚细胞定位，探讨重组人ECRG4（rhECRG4）蛋白的纯化和抑癌功能。方法用激光扫描共聚焦显微镜成像和Westernblot方法检测ECRG4蛋白的亚细胞定位；用噻唑蓝（MTT）方法检测rhECRG4蛋白的体外抑癌功能。结果激光扫描共聚焦显微镜成像显示，内源性和外源性ECRG4蛋白主要定位在细胞质。Westernblot方法在转染细胞的培养液中检测到ECRG4蛋白表达。rhECRG4蛋白浓度达到3mg／L时，抑制肿瘤细胞增殖；抑制率随着重组蛋白浓度增加而升高，呈剂量一效应关系（P〈0．05）。结论ECRG4蛋白可能是分泌蛋白；rhECRG4蛋白体外抑制肿瘤细胞增殖。

Objective To investigate the subcellular localization of human esophageal cancer re- lated gene 4 （ECRG4） protein, purification and tumor-suppressing function of recombinant human ECRG4 protein （rhECRG4） in esophageal carcinoma. Methods The laser-scanning confocal microscopy analysis and Western blotting assay were used to detect the subcellular localization of ECRC,4 protein. And methyl thiazol tetrazolium （MTI＇） assay was utilized to examine the tumor-suppressing function of rhECRG4 pro- tein. Results The images of laser-scanning confocal microscopy analysis indicated that endogenous and exogenous ECRG4 proteins were localized in the cytoplasm. And we also found ECRG4 protein in cellular medium by Western blot. The rhECRG4 protein in 3 mg/L concentration could inhibit tumor cells prolifera- tion in vitro, exhibiting dose-effect relationship （P 〈 0. 05 ）. Conclusion The human ECRG4 protein could be secretory protein. And the rhECRG4 protein inhibited the tumor cells growth in vitro.