hGM-CSF 基因转导的人胃癌、肝癌细胞研究

Study on hGM CSF modified human hepatoma and gastrocarcinoma cells

ｈＧＭ－ＣＳＦ是一种细胞因子，能通过多种机制激活机体免疫功能。为了研究ｈＧＭ－ＣＳＦ修饰的人肝癌、胃癌细胞的免疫功能的变化，将含该基因的逆转录病毒载体ＬＸＳＮ转染ＰＡ３１７包装细胞，经Ｇ４１８筛选较高滴度病毒分泌株（滴度为１．５×１０４ＣＦＵ／ｍｌ），用病毒上清感染肝癌（ＨＨＣＬ）和胃癌（ＭＫＮ４５），筛选后测ｈＧＭ－ＣＳＦ活性，最高分别为２７１Ｕ／１０６细胞·２４小时和２４３．８０Ｕ／１０６细胞·２４小时。ｈＧＭ－ＣＳＦ基因在肝、胃癌细胞中整合，未发生重排。ＭＨＣ分子测定结果显示经修饰的肝癌ＨＨＣＬ细胞ＭＨＣ分子的表达无改变，而修饰后胃癌细胞ＭＫＮ４５的ＭＨＣⅠ类分子表达明显增加。本实验为今后瘤苗研制提供了资料。

hGM CSF is a cytokine which can stimulate human immunosystem by various mechanisms.In order to study the biological behavior of hGM CSF modified tumor,we cloned the hGM CSF gene into LXSN retrovirus vector with which the PA317 cells was transducted.The maximal titre of retrovirus vector was 1.5×10 4CFU/ml.We used the virus suppernatant to infect human hepatoma cell line(HHCL) and gastrocarcinoma cell line (MKN45),respectively.The hGM CSF transducted cells were bioassayed.The maximal hGM CSF expression as 271U/10 6 cells/24h(HHCL hGM CSF) and 243.80U/10 6 cells/24h(MKN45 hGM CSF),respectively.The vectors were successfully intergrated into the genomic DNA of target cells.Both hGM CSF and neo gene were expressed.The FACS results demonstrated that increase in the expression of MHC Ⅰ was occurred in hGM CSF modified MKN 45 cells,however,no MHC expression change happened to hGM CSF modified HHCL cells.Our results could be utilized for future hGM CSF modified tumor vaccine research.