SV40T基因介导的永生化对细胞生物学特性的影响

The study of the biological characteristics of immortalized human ovarian carcinoma cells induced by SV40 T gene

目的研究SV40 T基因对其诱导的永生化细胞系细胞表型的影响。方法以手术切除的卵巢肉瘤样癌腹壁转移组织为材料,进行体外培养。将永生化基因———SV40 T抗原基因转染第10代细胞,经筛选,抗性克隆扩大培养,得到永生化细胞系。通过光学显微镜、电子显微镜、生长曲线测定、染色体分析、双层软琼脂培养、裸鼠接种、免疫组化等,研究转染与未转染细胞的生物学特性,通过四甲基偶氮唑蓝比色法(MTT)在相同条件下检测转染与未转染细胞对顺铂、阿霉素、足叶乙甙、拓扑替康四种化疗药物的敏感性。结果建立人卵巢肉瘤样癌细胞系,命名为BUPH:OVSC-1。经SV40 T抗原基因转染,建立人卵巢肉瘤样癌永生化细胞系,命名为BUPH:OVSC-2,两者均已长期稳定传代。通过比较,两者的生物学特性无明显差别,对不同化疗药物的敏感性无明显差别。结论SV40 T基因永生化的细胞保留了原细胞的分化表型,SV40介导的永生化细胞系可作为体外研究的模型。

Objective To study the biological characteristics of an immortalized human ovarian carcinoma cell line induced by SV40 T gene, and study the influence of SV40 T gene to the cell differentiation properties. Methods A specimen derived from metastatic tumor of a human ovarian sarcomatoid carcinoma was obtained and cultured in vitro. The 10th passage was transfected with SV40 T gene, colonies were isolated by G418 selection and expanded to cell lines. The morphology of the cells was observed by microscope and electromicroscope. Growth curve, karyotype analysis, culturing in soft agar, nude mice transplantation, immunohistochemistry etc were used to investigate its biological characteristics. The biological characteristics of transfected cells and its original ceils were compared. The sensitivities of two cells to cisplatin, adriamycin, etoposide and topotecan were tested in vitro respectively by MTT assay. Results An ovarian sarcomatoid carcinoma cell line BUPH：OVSC - 1 was established. After transfection and selection, an immortalized ovarian sarcomatoid carcinoma cell line BUPH：OVSC -2 was established. The two cell lines have been maintained for over 100 passages. Comparative studies revealed that the overall properties of the two cells were largely identical. No obvious differences were observed between the drug sensitivities of the two cell lines. Conclusion SV40 T gene - mediated immortalization can retain similar differentiation properties of their original cells. Immortalized cell lines induced by SV40 T gene can be useful cell models for in vitro research.