1.Establishment and identification of the gene cell line transfected aldose reductase like gene and its drug resistance to drugs containing carbonyl group.
Jian WANG ; Laxiang WAN ; Mengjin WU ; Zaiyang WAN ; Dangheng WEI ; Yongzong YANG
Chinese Journal of Hepatology 2002;10(6):445-448
OBJECTIVETo investigate the relationship between aldose reductase like protein (ARL-1) gene overexpressed in HCC cells and drug-resistance of the cell to drugs containing carbonyl group.
METHODSTo establish ARL-1 stable expression positive cell line, eukaryotic expression vectors containing ARL-1 gene cDNA were transfected into Hep cell mediated by lipofect AMINE. The positive monoclones were determined by PCR and RT-PCR, respectively. Then MTT assay was used to study the drug resistance ability of the cells to drugs containing carbonyl after incubating three days with those drugs.
RESULTSAfter ARL-1 gene transfection mediated by lipofect AMINE, one positive monoclonal cell overexpressing ARL-1 gene was selected. Compared with the control cell group, drug resistance ability of the positive cells to ADM and MMC which contain carbonyl group increased 2.3 and 3.17 fold, respectively (t=6.39, P=0.016 in ADM group and t=30.06, P=0.001 in MMC group). In the same time, drug resistance ability to 5-FU which has no carbonyl group had no statistical difference between positive monoclonal cell group and control cell group (t=0.684, P=0.531).
CONCLUSIONSThe Hep ARL-1 positive cell line with stable expression of ARL-1 gene has been established successfully and the up-regulation of ARL-1 gene may plays an important role in drug resistance of the cells to anticancer drugs containing carbonyl group.
Aldehyde Reductase ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; genetics ; physiology ; Fluorouracil ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Mitomycin ; pharmacology ; Transfection ; Tumor Cells, Cultured ; drug effects
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