宫颈永生化细胞和癌细胞的亚细胞蛋白质组学初步研究

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。

Subcellular Proteomics Analysis of Immortalized Cervical Cell and Cervical Cancer Cell

Objective To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from subcellular immortalized human endocervical cell （H8） and cervical cancer cell （Casld）, and to identify the differential expressions of subcellular proteins （cytoplasmic, membranous and nuclear proteins）. Methods H8 cells and Caski cells were incubated, and subcellular proteins of H8 cells and Caski cells were extracted and separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis （2-DE）. Then the selected differential protein spots were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and SWISS-PROT database. Results We obtained well-resolved, reproducible 2-DE patterns; 6 differentially expressed cytoplasmic proteins and 3 differentially expressed membranous proteins and 9 differentially expressed nuclear proteins were defined in 2-DE gels. Conclusions Subcellular proteins of cervical precancerous lesion and cervical cancer are separated and analyzed by means of 2-DE and MALDI-TOF-MS/MS. There are significant differences between H8 cells and Caski cells. These data may be valuable for research of cervical precancerous lesion and cervical cancer, or as diagnostic markers and therapeutic targets for cervical cancer.