β-葡萄糖醛酸苷酶基因转染人卵巢癌细胞的生物学特性

Biological characteristics of β-glucuronidase cDNA transfected ovarian cancer cell line 3AO-βG

目的 :将β 葡萄糖醛酸苷酶 (βG)基因转入卵巢癌细胞系 3AO ,建立高表达 βG的卵巢癌细胞模型 ,并观察转染细胞的生物学特性 .方法 :构建真核表达的逆转录病毒载体pcDNA3.1 βG ,利用阳离子脂质体将其转入人卵巢癌细胞株 ,用mRNA打点杂交、WesternBloting鉴定其转录与表达水平 ,并通过检测细胞生长、增殖及形态学的变化观察转染细胞的生物学特性 .结果 :转染阳性细胞 ,在mRNA与蛋白水平证实转染细胞内有 βG基因高表达 ;但转染前后细胞生长情况及细胞周期分析无显著性差别 .结论 :用脂质体介导法将βG基因在体外转染人卵巢癌 3AO细胞 ,经鉴定获得了生物学特性稳定的

AIM: To establish an ovarian carcinoma model with high expression of β glucuronidase by transfecting βG gene into the ovarian tumor 3AO cells and to study the biological characteristics of the transfected cells. METHODS: A eukaryotic expression vector pcDNA3.1 βG was constructed with βG cDNA and a eukaryotic expression vector pcDNA3.1(+). The pcDNA3.1 βG vector was transfected into ovarian cancer 3AO cells with lipofection reagent and the βG vector producer cell line 3AO/βG was isolated in 200 mg· L -1 nermycin medium. Transfection and isolation were confirmed by the means of RNA blotting and Western blot. The biological characteristics of the βG transfected cells were studied by the observation of their growth and proliferative period. RESULTS: It was confirmed that the βG gene had been stably integrated into the genomic DNA of the 3AO cells and highly expressed by the means of RNA blotting, immunohistochemistry and Western blot. No statistical difference was found between 3AO/βG and 3AO in the speed of growth and cell cycle. CONCLUSION: An ovarian carcinoma model with high expression of βG gene in vitro has been established, which can be used in future study on gene therapy.