差异显示cDNA转染DC细胞在肿瘤生物治疗中作用

The effect of dendritic cell transfected with differential displaying cDNA in biotherapy

目的通过差异显示技术筛选肿瘤细胞差异性基因并转染到树突状细胞中,验证转染后DC所诱导的CTL杀伤效能。方法将Lewis肺癌细胞种植在C57BL/6J小鼠后,取肿瘤组织和正常组织进行差异显示反转录PCR技术分析鉴定,得到的差异显示基因经测序后,构建pcDNA3.1-DD-cDNA质粒,经脂质体转染至未成熟树突状细胞中。分别通过体内体外实验检验树突状细胞诱导的细胞毒T细胞的杀伤活性。结果筛选出Lewis肺癌细胞的一条DD-cDNA的长度为730 bp,通过与Genbank序列数据库进行同源性比对,未发现同源性信息,为一全新基因。体内试验和体外试验中cDNA转染组CTL对肿瘤细胞杀伤效果均明显强于单纯肿瘤裂解物刺激组。结论 Lewis肺癌细胞表达一条肿瘤特异性基因,将其转染到树突状细胞中后,可以明显增强树突状细胞诱导的CTL的杀伤活性。

[ Objective ] To select the differential displaying gene of tumor, then transfect it to dendritic cell （DC）. To test the killing activity of cytotoxic T lymphocyte induced by transfected dendritic cell. [ Methods ] Tumorous and normal tissues, taken from C57BI/6J mice planted with Lewis lung carcinoma, were analyzed by differential display PCR. The differential displaying gene was sequenced and constructed the pcDNA3.1-DD-cDNA plasmid which was transfected into immature dendritic cell. The killing activity of cytotoxic T lymphocyte （CTL） induced by transfected DC were tested in vivo and in vitro separately. [Results] A DD-cDNA which was 730 bp was identified, homology of its sequences was not found compared to Genbank. The killing activity of CTL induced by transfected dendritic cell was dramatically stronger than that induced by DC stimulated by tumor cell lysate. [ Conclusion ] A differential displaying gene expresses in Lewis lung carcinoma cell, it can dramatically improve the CTL induced by dendritic transfected with this gene.