B_7基因转染联合LAK细胞治疗肝癌的体外研究

Experimental study on B_7 gene transfection combined with LAK-cells for liver carcinoma in vitro

目的 :探讨体外诱导的 L AK细胞对 B7基因转染的肝癌细胞的杀伤作用。方法 :构建含人 B7- 1 基因 c DNA的真核表达载体 p RCCMV- B7- 1 ,脂质体介导转染法将 p RCCMV- B7- 1 转入人肝癌细胞系 SMMC772 1 ,G4 1 8筛选转染肝癌细胞 (B7+ SMMC772 1 )。RT- PCR、流式细胞仪鉴定目的基因表达。MTT比色试验体外检测 L AK细胞对 B7- 1 转染肝癌细胞的杀伤活性。结果 :B7+ SMMC772 1细胞稳定表达 B7基因。L AK细胞对 B7+ SMMC772 1细胞的杀伤活性明显高于野生型 SMMC772 1者 ,结果有统计学意义。结论 :B7基因体外转染人肝癌细胞后可表达有活性的 B7分子 ,L AK细胞对表达 B7- 1

ObjectiveTo study the killing effects of induced LAK cells on B 7 gene transfected cells of liver carcinoma. MethodsA new vector pRCCMV B 7 1 containing B 7 1 gene was constructed to establish the new cell line B 7 +SMMC7721 through transfection B 7 gene into the hepatocarcinoma cell SMMC7721 by liposome mediated transduction. RT PCR and flow cytometry were used to identify the expression of the transfected gene. MTT assay was used to evaluate the killing effects of LAK cells activated by IL 2 on the transfected line and the wild type of the cells. ResultsB 7 gene was steadily expressed in B 7 +SMMC7721 and LAK cells had effectively killing effects on the B 7 +SMMC7721 cells. ConclusionB 7 gene can be transfected into hepatocarcinoma cells and can be expressed steadily in vitro , which can increase the efficiency of LAK cells activated by IL 2 on them.