摘要
目的 研究腺病毒介导的多种基因在肝癌细胞中的表达和对肝癌细胞生长的影响。方法 构建含人p5 3、B7 1、GM CSF和IL 2 4种目的基因的重组腺病毒载体 ,感染 3种肝细胞癌细胞系和肝细胞系L0 2 ,运用ELISA、免疫组化、流式细胞仪等方法 ,检测目的基因在肝癌细胞中的表达 ,和对肝癌细胞生长的抑制及诱导其凋亡的作用。结果 肝癌细胞对腺病毒高度易感 ,腺病毒多重感剂量为 5 0 (5 0MOI)时 ,可使 90 %左右的肝细胞癌表达目的基因。除IL 2外 ,其他 3个目的基因都能在肝癌细胞中高效表达。导入多基因后 ,肝癌细胞的生长受到一定程度的抑制 ,并诱导肝癌细胞发生凋亡 ;而L0 2肝细胞的生长却未受影响。结论 腺病毒载体是进行肝癌基因治疗的理想载体 ,由一个腺病毒载体介导的多种目的基因 ,能在肝癌细胞系中高效表达 ,且能抑制肝癌细胞的生长和诱导其调亡。
Objective To investigate the expression of multigenes mediated by adenovirus in liver cancer cells and the effects on growth of cells transducted with multigenes. Methods By construction of recombinant adenovirus containing human p53, B7-1, GM-CSF, and IL-2 genes (Ad-multigenes), the expression level of target genes in three human hepatocellular carcinoma cell lines and a human hepatocellular cell line L02 was detected using ELISA, immunohistochemistry and FACS assay and the change of growth of these cells and the tumor cell apoptosis were observed. Results The human hepatic cells and liver cancer cells were all sensitive to adenovirus infection. At a MOI of 50 PFU/cell, among the cells examined nearly 90% were positive expression and except IL-2, other three genes were expressed with high efficiency. The growth of Ad-multigenes-transduced liver cancer cell lines was inhibited and apoptosis was induced, but the growth of Ad-multigenes-transduced normal hepatic cell line L02 did not change. Conclusion These results indicate that the adenovirus is an efficient vector for gene transfer into human liver cancer cells. These liver cancer cell lines transduced with multigenes constructed on one recombinant adenoviral vector can highly express target genes and their growth was inhibited, and apoptosis appeared.
出处
《中国普外基础与临床杂志》
CAS
2000年第6期357-359,361,共4页
Chinese Journal of Bases and Clinics In General Surgery
关键词
肝细胞癌
腺病毒
多基因
基因治疗
细胞凋亡
Hepatocellular carcinoma Adenovirus Multigenes Gene therapy Apoptosis
