IL-2基因转导小鼠肝癌细胞的实验研究

Study of Mouse Hepatoma Cells Transfected with IL-2 Gene

用重组人白细胞介素-2(IL-2)含信号肽cDNA的缺陷型逆转录病毒,将IL-2基因转导入小鼠H_(22)肝癌细胞株,经PCR和RT-PCR检测证实了转导后的细胞DNA内确有NeoR和IL-2基因的存在的表达。分别用光镜和电镜观察了经IL-2基因转录的H_(22)细胞体外形态,MTT法检测细胞体外增殖能力以及皮下接种H_(22)-IL-2细胞后观察肿瘤生长能力。结果表明经IL-2基因转导的鼠肝癌细胞体外增殖能力没有明显变化,但体内致瘤性却显著下降,丝裂霉素处理后作为瘤苗接种能抑制其后野生型H_(22)细胞的再次攻击。本研究为制备新型瘤苗打下了基础。

Application of interliukin-2 (IL-2) has already been promising in treating tumors, Weinvestigeted the effect of IL-2 on mouse hepatoma. Methods: Retrovirl vector was used to introduced the IL-2 gene into H22 cells, a mouse hepatoma cell line. The IL-2 gene and marker gene (NeoR) were assessed by using PCR and RT-PCR methods. Results: The results demonstrated that NeoR gene and IL-2 gene have been inserted and satisfactorily expressed. In vitro the transduced cells showed no obyious changes in morphology and growth ability, but its tumorigenicity decreased in vivo. Inoculation of H22-IL-2 positive cell vaccine to mice resulted in mice resistance to the rechallenge of parental H22 cells. IL-2 gene-transfected H22 cells were inactiveted with mitomycin C (0.4mg/ml, 3 hours at 37℃) to serve as vaccine. Mice were immunized i.p. three times at 7-day intervals, with 2 × 106 cells. Conclusion: The vaccine transfected with IL-2 gene had anti-tumor ability. The expression of IL-2 in H22 cells can reduce their tumorigenicity and/or metastatic potential and increase the recognition of tumor by the host immune system.