人γ-干扰素基因导入人大肠癌细胞中的方法及鉴定

The Method and Identification of Human Interferon-γGene Transfered into Colon Cancer Cell

目的：为将人γ－干扰素（ｈｕＩＦＮ－γ）基因尽快用于临床基因治疗，寻求基因导入的有效方法。方法：利用阳离子脂质体Ｌｉｐｏｆｅｃｔａｍｉｎｅ将ｈｕＩＦＮ－γ基因导入人大肠癌细胞株，利用ＰＣＲ、ＲＴ－ＰＣＲ对该基因的整合和表达进行鉴定，并对细胞培养上清中γ－干扰素的活性进行了检测。结果：该基因已经有效地进入细胞内并实现了表达。结论：利用脂质体可以有效地将ｈｕＩＦＮ－γ基因导入肿瘤细胞内，基因表达的持续时间可以满足肿瘤基因治疗的需要。不同细胞株其上清中ｈｕＩＦＮ－γ的分泌量不同。

Objective:In order to discover the available mode on interferon-γ gene therapy. Methods:The whole length of huIFN-γ cDN A was subcloned into expression vector pcDNA-3 after sequence mensuration, and then transduced to human colon cancer cell line LOVO, SW620, HCT 116BG mediated b y Lipofectamine. The integration and expression of huIFN-γ gene were verified by PCR and RT-PCR and the huIFN-γ activity in supernatant fluid of cell culture dish was measured by ELISA. Results:huIFN-γ gene was successful int egrated into human colon cancer cell and availably expressed. It gave useful inf ormation to develop the cytokine gene therapy in colon cancer. Conclusion: The huIFN-γ gene be effectively transduced into human colon cancer cell. The expression time of huIFN-γ maybe meet the demand of gene therapy and the qua ntity of huIFN-γ in the supernatant fluid of different cell lines is variant.