一种新的肿瘤相关蛋白FAM92A1-289的原核表达、纯化及其在肾细胞癌中的特性研究
Prokaryotic expression, purification of a new tumor-relative protein FAM92A1-289 and its characterization in renal cell carcinoma.
作者信息
Liang Shufang, Gong Fengming, Zhao Xinyu, Wang Xianhuo, Shen Guobo, Xu Yuhuan, Yang Hanshuo, Ruan Xuzhi, Wei Yuquan
机构信息
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, People's Republic of China.
出版信息
Cancer Lett. 2009 Apr 8;276(1):81-7. doi: 10.1016/j.canlet.2008.10.043. Epub 2008 Dec 6.
In order to study the characterization of a new tumor-relative FAM92A1-289 protein, we first constructed plasmid FAM92A1-pQE30 for fusion expression in Escherichia coli. The recombinant protein FAM92A1-289 was affinity-purified by Ni2+-charged resin and separated by HPLC chromatography with high purity, and it was further identified by electrospray ionization-mass spectrometry. Furthermore, the expression and cell localization of FAM92A1-289 by immunohistochemistry using our self-prepared polyclonal antibody showed it was expressed in cytoplasm of renal carcinoma. FAM92A1-289 mRNA was expressed in 2 of 10 kidney tissues and in 6 of 12 primary renal tumors. FAM92A1-289 can promote cell growth in vitro and in vivo by colony formation and mouse xenograft assay. Our present data indicated FAM92A1-289 is a new tumor-related gene with oncogenic potentials to probably play roles in renal carcinogenesis.
为了研究一种新的肿瘤相关蛋白FAM92A1-289的特性，我们首先构建了质粒FAM92A1-pQE30用于在大肠杆菌中融合表达。重组蛋白FAM92A1-289通过镍离子负载树脂进行亲和纯化，并通过高效液相色谱法分离得到高纯度产物，进一步通过电喷雾电离质谱法进行鉴定。此外，使用我们自行制备的多克隆抗体通过免疫组织化学检测FAM92A1-289的表达和细胞定位，结果显示其在肾癌细胞质中表达。FAM92A1-289 mRNA在10例肾组织中的2例以及12例原发性肾肿瘤中的6例中表达。通过集落形成和小鼠异种移植实验，FAM92A1-289在体内外均可促进细胞生长。我们目前的数据表明FAM92A1-289是一个新的肿瘤相关基因，具有致癌潜能，可能在肾癌发生过程中发挥作用。
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