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目的通过蛋白质双向凝胶电泳方法(2-DE)建立人类乳头状瘤病毒-16阳性的宫颈上皮永生化细胞(H8细胞株)和宫颈癌细胞(Caski细胞株)亚细胞(细胞质、细胞膜、细胞核)蛋白电泳图谱,进行差异蛋白质组学研究筛选差异蛋白点。方法分别对H8和Caski细胞提取细胞质、细胞膜和细胞核蛋白,通过2-DE筛选出差异点进行基质辅助激光解吸飞行时间串联质谱(MALDI-TOF-MS/MS)分析。结果对H8细胞和Caski细胞亚细胞蛋白双向电泳图谱比较,初步比较筛选了6种细胞质差异蛋白点、3种细胞膜差异蛋白点和9种细胞核差异蛋白点。结论通过对HPV-16阳性的宫颈癌前和癌变的亚细胞结构差异蛋白进行分析,为寻找宫颈癌特异性肿瘤标志物和靶向治疗提供实验依据。
Objective To establish human papillomavirus-16 positive cervical epithelial immortalized cells (H8 cell line) and cervical cancer cell (Caski cell line) subcellular (cytoplasm, cell membrane, nucleus by protein two-dimensional gel electrophoresis method ) Protein electrophoresis pattern, differential proteomics study screening of differential protein spots. Methods Cytoplasm, cell membrane and nuclear protein were extracted from H8 and Caski cells, respectively. The differences were screened by 2-DE and analyzed by matrix-assisted laser desorption / time of flight tandem mass spectrometry (MALDI-TOF-MS / MS). Results Compared with the two-dimensional gel electrophoresis of H8 cells and Caski cells, six kinds of cytoplasmic differential proteins spots, three kinds of cell membrane differential proteins spots and nine kinds of nuclei differential protein spots were screened and compared. Conclusion The HPV-16 positive cervical precancerous and cancerous subcellular structural proteins were analyzed to provide experimental evidence for finding specific cervical cancer-targeted tumor markers and targeted therapy.