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Effects of multidrug resistance, antisense RNA on the chemosensitivity of hepatocellular carcinoma cells

Hepatobiliary & Pancreatic diseases International 2006年第4期5卷 552-559页

作者： Bo Li, Tian Ye, Lei Zhao, De-Hua Li, Xing-Hua Gou, Lan-Ying Zhao, Lei Han, Lin Chen, Lu-Nan Yan and Jian-Ping Gong department of Hepatobiliary surgery, Seco.d Affiliated hospital, Chongqing university of Medical Sciences, Chongqing 400010, china department of Traditional Chinese surgery, Seco.d Affiliated hospital, Luzhou Medical co.lege, Luzhou 646000, china genetic engineering laboratory, chengdu di’ao group co., ltd., chengdu 610041, china department of general surgery, west china hospital, sichuan university, chengdu 610041, china department of Hepatobiliary surgery Second Affiliated Hospital Chongqing University of Medical Sciences Chongqing 400010 China.

BACKGROUND: Multidrug resistance is a major obstacle in cancer chemotherapy. We examined whether the antisense RNA of multidrug resistance gene 1 (mdr1) co.ld reverse multidrug resistance in the human hepatocellular carcinoma (HCC) cell line SMMC7721/ADM. METHODS: The reco.binant adenoviruses pAdEasy- GFP-ASmdr1 product was produced by the adenoviral vector AdEasy system, which can express antisense RNA against the mdr1 gene. Following that, the reco.binant adenovirus was transfected into the P-glyco.rotein- producing multidrug resistance cell line, SMMC7721/ADM human HCC cells resistant to adriamycin (ADM) and daunorubicin (DNR). In order to investigate the reversal of multidrug resistance phenotype, we measured the expression of mdr1 mRNA by RT-PCR and the production of P-glyco.rotein by flow cytometry. The sensitivities for ADM and DNR SMMC7721/ADM cells were examined by [3-(4, 5-dimethylthi-azol-2-yl)-2,5 diphenyl-terazolium bromide] (MTT) analysis. RESULTS: The low-level expression of mdr1 mRNA and P-glyco.rotein production were observed in parental sensitive cells SMMC/7721 in addition to the overexpressionof mdr1 mRNA and P-glyco.rotein in SMMC7721/ADM cells. The transfection of antisense-RNA into SMMC7721/ ADM cells resulted in decreases of mdr1 mRNA and P-glyco.rotein, but increase of drug sensitivities. The sensitivities of transfected SMMC7721/ADM cells to ADM and DNR in IC50 reduced by 31.25% and 62.96% respectively. co.CLUSIONS: Mdr1 antisense RNA can increase the sensitivities of SMMC7721/ADM cells to anticancer drug by decreasing the expression of the mdr1 gene and inhibiting P-glyco.rotein expression. This strategy may be applicable to cancer patients with P-glyco.ortein mediated multidrug resistance.

关键词： antisense nucleic acid multiple drug resistance P-glyco.rotein

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