[Improvement of combination chemotherapy tolerance of human umbilical cord blood CD(34)(+) cells transducted with double drug resistance genes by a bicistronic retroviral vector]

[Improvement of combination chemotherapy tolerance of human umbilical cord blood CD(34)(+) cells transducted with double drug resistance genes by a bicistronic retroviral vector] Research Abstract Details 

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[Improvement of combination chemotherapy tolerance of human umbilical cord blood CD(34)(+) cells transducted with double drug resistance genes by a bicistronic retroviral vector] Abstract Text:

J Wang,X Xia,Z Chen,D Lu,J Xue,C Ruan,

OBJECTIVE: To explore whether human umbilical cord blood hematopoietic progenitor cells transduced with human aldehyde dehydrogenase class 3 (ALDH3) and multidrug resistance gene (MDR1) could increase resistance to 4-hydroxycyclophosphamide (4-HC) and P-glycoprotein effluxed drugs. METHODS: A bicistronic retroviral vector G1Na-ALDH3-IRES-MDR1 cDNA was constructed and transfected the packaging cell lines GP + E86 and PA317 by LipofectAMINE method, using the medium containing VCR and 4-HC for cloning selection and ping-ponging supernatant infection between ecotropic producer clone and amphotropic producer clone, cord blood CD(34)(+) cells were enriched with a high-gradient magnetic cell sorting system (MACS), and then repeatedly transfected with supernatant of retrovirus containing human ALDH3 and MDR1 cDNA under stimulation of hematopoietic growth factors. PCR, RT-PCR, Southern blot, Northern blot, FACS and MTT assay were used to evaluate the transfection and expression of the double genes. RESULTS: The purity of cord blood CD(34)(+) cells was approximately 91% and the recovery rate was 72%. The highest titer of recombinant amphotropic retrovirus in the supernatant was up to 6.5 x 10(5) CFU/ml. The efficiency of gene transduction was 18%, 20% and 16.7% tested by colony formation, PCR and FACS, respectively. Rhodamine 123 efflux showed 16% transduced cells with P-gp function. No helper virus was found by both nested PCR and rescue assay. The MTT analysis showed a 3.5 to 6.8-fold increase of resistance of transducted cells to cyclophosphamide and P-glycoprotein effluxes drug as compared with the nontransduced cells. CONCLUSION: The efficiency and co-expression of this dual genes transfer system provided a foundation for ameliorating combination chemotherapy toxicity in clinical trial.

[Improvement of combination chemotherapy tolerance of human umbilical cord blood CD(34)(+) cells transducted with double drug resistance genes by a bicistronic retroviral vector] Publishing Authors By Initials

J Wang,X Xia,Z Chen,D Lu,J Xue,C Ruan,

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[Improvement of combination chemotherapy tolerance of human umbilical cord blood CD(34)(+) cells transducted with double drug resistance genes by a bicistronic retroviral vector] Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Zhonghua xue ye xue za zhi = Zhonghua xueyexue zaz

VOLUME: 22

Page Numbers: 197-201

Journal Abbreviation: Zhonghua Xue Ye Xue Za Zhi

ISSN: 0253-2727

DAY: 5

MONTH: Apr

YEAR: 2001

[Improvement of combination chemotherapy tolerance of human umbilical cord blood CD(34)(+) cells transducted with double drug resistance genes by a bicistronic retroviral vector] Information

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LANGUAGE: chi

NlmUniqueID: 8212398

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AFFILIATION: Jiangsu Institute of Hematology, First Affiliated Hospital of Suzhou Medical College, Suzhou 215006, China.

Country: China

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MEDLINETA: Zhonghua Xue Ye Xue Za Zhi

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