卵巢癌细胞中乳源免疫调节肽受体蛋白的筛选

Screening of immunomodulating peptide receptor protein from human ovarian carcinoma cells

目的乳源免疫调节肽(PGPIPN)在人卵巢癌细胞株(SKOV3)上进行受体蛋白的筛选。方法异硫氰酸荧光素(FITC)标记PGPIPN,作用于SKOV3观察其细胞定位;从SKOV3中提取作用部位的蛋白;通过NHS-activated亲和层析柱进行分离和纯化受体蛋白质,SDS-PAGE的初步鉴定。结果在激光扫描共聚焦显微镜下观察荧光标记肽结合于SKOV3的胞膜上。通过提取膜蛋白和亲和层析法成功筛选了受体膜蛋白。结论 PGPIPN的作用位点位于细胞膜上,其抗癌作用可能通过胞膜结合蛋白而启动信号转导通路,为进一步探讨其抗癌机制奠定基础。

Objective To screen receptor protein of immunomodulating peptide （PGPIPN） in ovarian cancer cell lines SKOV3. Methods PGPIPN was labelled fluorescein isothiocyanate （ FITC ）, whose binding site at the cell was observed under laser scanning confocal microscope. The whole membrane proteins were extracted from ovarian cancer cell by membrane protein extraction kit. Taking PGPIPN as ligand, the receptor protein was screened and purified by NHS-activated affinity chromatography, which were preliminary identificated by SDS-PAGE. Results Compared with the control, the fluorescence labeled peptides were observed at the cell membrane by laser scanning confocal microscope. The membrane proteins were successful extracted by using reagent kit. Receptor protein had been screened and purified from membrane proteins by using affinity chromatography. Conclusion PGPIPN receptor locates at the cell membrane. The anticancer of PGPIPN may effect through binding membrane protein to trigger signal transduction. This study has laid the foundation for further probing the anticancer mechanism of PGPIPN.