| 摘要: |
| 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。 |
| 关键词: 卵巢肿瘤 HO8910PM细胞株 绿色荧光蛋白质类 转染 |
| DOI：10.3724/SP.J.1008.2012.00950 |
| 投稿时间：2012-04-27修订日期：2012-07-17 |
| 基金项目:国家重点基础研究发展计划(“973”计划,2011CB707902). |
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| Establishment of a fluorescence-labeled human ovarian cancer cell line |
| ZHANG Xiao-mei,FANG Liao-qiong,WANG Zhi-biao* |
(Department of Biomedical Engineering, Chongqing Medical University, Chongqing 400016, China
*Corresponding author.) |
| Abstract: |
| ObjectiveTo establish a human ovarian cancer cell line stably expressing enhanced green fluorescent protein(EGFP), so as to carry out visualized research on whole ovarian cancer. MethodsWe transfected human ovarian cancer cell line HO8910PM by gene transfection, and obtained cells stably expressing EGFP by sub-cloning amplification and selection with G418-resistance.The expression rate of EGFP was analyzed by flow cytometry (FC).The growth curve, adhesion, migration and invasion experiments were employed to study the biological behaviors of the cells transfected with EGFP. ResultsFlow cytometry results showed that EGFP positive rate of screened EGFP-HO8910PM cells was higher than 99%. The cell growth, adhesion, invasion and migration abilities of cells were not significantly changed after transfection. ConclusionWe have successfully established a cell line EGFP-HO8910PM stably expressing EGFP and at mean time maintaining the characteristics of the parent cell line, which lays a foundation for whole-body visualization research of human ovarian cancer in vivo. |
| Key words: ovarian neoplasms HO8910PM cell line green fluorescent proteins transfection |
