转白细胞介素2基因人肝癌细胞株的建立及其部分生物活性的研究

Establishment of a human liver cancer cell line transfected with IL-2 cDNA and its biologic aclivity.

目的建立转白细胞介素2(IL-2)基因人肝癌细胞株,并研究IL-2的表达情况及其在动物体内的生物活性。方法用重组载体逆转录病毒LNC-IL_2与肝癌细胞SMMC-7721混合培养、G_(418)筛选,建立转基因细胞株。以斑点杂交法、RT-PCR法及MTT法分别从DNA、RNA及蛋白质水平检测外源性IL-2基因。同时在裸鼠皮下进行致瘤及抑瘤试验。结果及结论 IL-2基因成功地整合到SMMC-7721细胞基因组中,且持续大量表达,高表达持续88天仍无降低趋势;裸鼠皮下接种转基因细胞发现致瘤性明显减弱,且能在一定程度上诱导体内的抗瘤作用。

Objective To obtain IL-2 gene transfected human liver cancer cells and study IL-2 expression and its biologic activity in vivo. Methods Human liver cancer cells SMMC-7721 were cocultured with recombinant retroviral vector LNC-IL-2, and screening was performed in G418 medium. The exogenous IL-2 cDNA at the DNA, RNA, and protein levels were determined by using dot hybridization, PR-PCR and MTT methods respectively. The tumorigenesis and antitumorigenesis of the screened liver cancer cell with subcutaneous injection in nude mice were observed. Results and Conclusion The IL-2 cDNA was successfully integrated into SMMC-7721 cell genomic DNA and highly and continuously expressed for more than 88 days. Subcutaneous vaccination of the nude mice with transfected cells revealed an obvious suppression of its tumorigenictiy, and could induce antitumor activity in vivo.