蛋白质酵母双杂交系统的建立及其在大肠癌肝转移研究中的意义

目的 建立酵母双杂交系统,筛选与FasL相互作用的蛋白,探讨FasL与大肠癌肝转移的关系。方法 以FasL基因构建诱饵蛋白质粒,筛选人胎肝cDNA文库,鉴定与FasL相互作用的蛋白,通过生物信息学分析FasL及其相互作用蛋白在大肠癌肝转移中的作用。结果 筛选出10个与FasL特异性相互作用的蛋白,包括金属硫蛋白1K、1G、2A,组织蛋白酶B,脂肪酸合成酶,干扰素α诱导蛋白27,磷脂清除酶,丝氨酸／苏氨酸样激酶,锚着黏附蛋白以及纤维微丝蛋白-5等。结论 成功建立了筛选FasL相互作用蛋白的酵母双杂交系统,并初步证明FasL与组织蛋白酶、金属硫蛋白、锚着黏附蛋白等之间的相互作用与大肠癌肝转移密切相关。

The construction of yeast two-hybrid method in the protein-interactions and its significance in hepatic metastasis of colorectal carcinoma

OBJECTIVE: To construct the yeast two-hybrid system, and screen the proteins which interact with FasL, and investigate the relationship of FasL and hepatic metastasis of colorectal carcinoma. METHODS: We have cloned the FasL gene into the pGBKT7 vector as the bait, then screened the fetal liver cDNA library, and have got a series of specific proteins that interact with FasL protein. Using the bioinformatics, we analyzed the interacting proteins in the mechanism of hepatic metastasis of colorectal carcinoma. RESULTS: We have screened several proteins that interaction with FasL protein, including metallothionein 1K, 1G, 2A, cathepsin B, fatty acid synthase, interferon alpha-inducible protein 27, phospholipid scramblase, Ser/Thr-like kinase, anchor attachment protein, fibulin-5. CONCLUSIONS: We have successfully constructed the yeast two-hybrid system, and preliminary identified that the interaction between FasL, metallothionein, cathepsin and anchor attachment protein is radically related to the hepatic metastasis of colorectal carcinoma.