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Titolo:
A bicistronic retroviral vector to introduce drug resistance genes into human umbilical cord blood CD34(+) cells to improve combination chemotherapy tolerance
Autore:
Wang, JS; Chen, ZX; Xia, HM; Lu, DR; Xue, JL; Ruan, CG;
Indirizzi:
Suzhou Med Coll, Affiliated Hosp 1, Jiangsu Inst Hematol, Suzhou 215006, Peoples R China Suzhou Med Coll Suzhou Peoples R China 215006 ou 215006, Peoples R China Fudan Univ, Inst Genet, Shanghai 200433, Peoples R China Fudan Univ Shanghai Peoples R China 200433 nghai 200433, Peoples R China
Titolo Testata:
CHINESE MEDICAL JOURNAL
fascicolo: 1, volume: 114, anno: 2001,
pagine: 25 - 29
SICI:
0366-6999(200101)114:1<25:ABRVTI>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
MDR1; COEXPRESSION;
Keywords:
gene therapy aldehyde-dehydrogenase-class1 gene (ALDH-1); multidrug resistance gene (MDR1); retrovirus vector; gene co-expression; hematopoietic stem cells; cord blood;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Citazioni:
11
Recensione:
Indirizzi per estratti:
Indirizzo: Wang, JS Suzhou Med Coll, Affiliated Hosp 1, Jiangsu Inst Hematol, Suzhou 215006, Peoples R China Suzhou Med Coll Suzhou Peoples R China 215006 , Peoples R China
Citazione:
J.S. Wang et al., "A bicistronic retroviral vector to introduce drug resistance genes into human umbilical cord blood CD34(+) cells to improve combination chemotherapy tolerance", CHIN MED J, 114(1), 2001, pp. 25-29

Abstract

Objective To study whether human umbilical cord blood CD34(+) cells transduced with human aldehyde dehydrogenase class-1 (ALDH-1) and multidrug resistance gene (MDR1) have increases resistance to 4-Hydroperoxycyclo-phosphamide (4-HC) and P-glycoprotein effluxed drugs. Methods A bicistronic retroviral vector G1Na-ALDH1-IRES-MDR1 was constructed and used to transfect the packaging cell lines GP + E86 and PA317 by LipofectAMINE method, using the medium containing VCR and 4-HC agents for cloning selection and ping-ponging supernatant infection between the ecotropic producer clone and the amphotropic producer clone, we obtained high titer amphotropic PA317 producing cells with high titers up to 5.6 x 10(5) CFU/ml. Cord blood CD34+ cells were transfected repeatedly with supernatant of retrovirus containing human ALDH-1 and MDR1cDNA under the stimulation of hemopoietic growth factors. Results Bicistronic retroviral vector construction was verified by restriction endonuclease analysis. Polymerase chain reaction (PCR), reverse transcription (RT)-PCR, Southern blot, Northern blot, fluorescenceactivated cell sorting (FACS) method and 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analyses showed that dual drug resistance genes have been integrated into the genomic DNA of cord blood CD34+ cells and expressed efficiently. The transgenes recipient cells confered 4-fold stronger resistance to 4-HC and 5.5 to 7.2-fold P-glycoprotein effluxed drug than untransduced cells. Conclusion The bicistronic retroviral vector-mediated transfer of two different types of drug resistance genes into human cord blood CD34(+) cells and co-expression provided an experimental foundation for improving combination chemotherapy tolerance in tumor clinical trial.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 03/12/20 alle ore 15:35:32