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反义核酸(antisense nucleic acid)

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中华预防医学杂志 2003年第5期37卷 67-67页

作者：方福德中国医学科学院基础医学研究所 100005北京

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

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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, Second Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400010, China Department of Traditional Chinese Surgery, Second Affiliated Hospital, Luzhou Medical College, 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) could reverse multidrug resistance in the human hepatocellular carcinoma (HCC) cell line SMMC7721/ADM. METHODS: The recombinant 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 recombinant adenovirus was transfected into the P-glycoprotein- 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-glycoprotein 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-glycoprotein production were observed in parental sensitive cells SMMC/7721 in addition to the overexpressionof mdr1 mRNA and P-glycoprotein in SMMC7721/ADM cells. The transfection of antisense-RNA into SMMC7721/ ADM cells resulted in decreases of mdr1 mRNA and P-glycoprotein, 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. CONCLUSIONS: 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-glycoprotein expression. This strategy may be applicable to cancer patients with P-glycoportein mediated multidrug resistance.

关键词： antisense nucleic acid multiple drug resistance P-glycoprotein

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Gene therapeutic approaches to inhibit hepatitis B virus replication

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World Journal of Hepatology 2015年第2期7卷 150-164页

作者： Maren Gebbing Thorsten Bergmann Eric Schulz Anja Ehrhardt Institute of Virology and Microbiology Center for Biomedical Education and Research(ZBAF)Department of Human MedicineFaculty of HealthUniversity Witten/Herdecke58453 WittenGermany

Acute and chronic hepatitis B virus(HBV) infections remain to present a major global health problem. The infection can be associated with acute symptomatic or asymptomatic hepatitis which can cause chronic inflammation of the liver and over years this can lead to cirrhosis and the development of hepatocellularcarcinomas. Currently available therapeutics for chronically infected individuals aim at reducing viral replication and to slow down or stop the progression of the disease. Therefore, novel treatment options are needed to efficiently combat and eradicate this disease. Here we provide a state of the art overview of gene therapeutic approaches to inhibit HBV replication. We discuss non-viral and viral approaches which were explored to deliver therapeutic nucleic acids aiming at reducing HBV replication. Types of delivered therapeutic nucleic acids which were studied since many years include antisense oligodeoxynucleotides and antisense RNA, ribozymes and DNAzymes, RNA interference, and external guide sequences. More recently designer nucleases gained increased attention and were exploited to destroy the HBV genome. In addition we mention other strategies to reduce HBV replication based on delivery of DNA encoding dominant negative mutants and DNA vaccination. In combination with available cell culture and animal models for HBV infection, in vitro and in vivo studies can be performed to test efficacy of gene therapeutic approaches. Recent progress but also challenges will be specified and future perspectives will be discussed. This is an exciting time to explore such approaches because recent successes of gene therapeutic strategies in the clinic to treat genetic diseases raise hope to find alternative treatment options for patients chronically infected with HBV.

关键词： Gene therapy Hepatitis B virus antisense nucleic acid RNA interference Designer nuclease Ribozyme DNAzyme Dominant negative mutant External guide sequence DNA vaccination

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