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目的建立稳定高表达增强型绿色荧光蛋白(EGFP)的人卵巢癌细胞株。方法采用基因转染的方法,将EGFP基因导入人卵巢癌细胞HO8910PM中,通过G418筛选、亚克隆扩增获得稳定表达绿色荧光蛋白的EGFP-HO8910PM细胞株,并用流式细胞术检测所获细胞株的EGFP表达率,通过细胞生长曲线、黏附实验、侵袭迁移实验比较EGFP-HO8910PM细胞和HO8910PM细胞的生物学行为。结果筛选出的EGFP-HO8910PM细胞经流式细胞仪检测EGFP阳性表达率达99%以上,EGFP-HO8910PM细胞和HO8910PM细胞生长、黏附及侵袭迁移能力无统计学差异。结论成功建立了稳定表达EGFP且保持母株细胞特性的人卵巢癌细胞株EGFP-HO8910PM,为人卵巢癌整体活体应用中可视化研究打下基础。
Objective To establish a human ovarian carcinoma cell line stably expressing EGFP. Methods EGFP gene was transfected into HO8910PM cells by gene transfection. EGFP-HO8910PM cells stably expressing green fluorescent protein (EGFP-HO8910PM) were screened by G418 and subcloned into EGFP-HO8910PM cells. The cell lines were obtained by flow cytometry The EGFP expression rate of EGFP-HO8910PM cells and HO8910PM cells were compared by cell growth curve, adhesion assay and invasion and migration assay. Results EGFP-HO8910PM cells screened by flow cytometry showed that the positive rate of EGFP was over 99%. There was no significant difference in the growth, adhesion, invasion and migration of EGFP-HO8910PM cells and HO8910PM cells. Conclusion The human ovarian cancer cell line EGFP-HO8910PM, which stably expresses EGFP and maintains the characteristics of the mother plant, has been successfully established and laid the foundation for the visualization of human ovarian cancer in vivo.