Format

Send to

Choose Destination
Acta Pharmacol Sin. 2001 Oct;22(10):949-55.

Genetic modification of hematopoietic progenitor cells for combined resistance to 4-hydroperoxycyclophosphamide, vincristine, and daunorubicin.

Author information

1
Department of Hematology, the Affiliated Hospital of Guiyang Medical College, Guiyang 550004,China. jswang@shmu.edu.cn

Abstract

AIM:

To investigate whether human peripheral blood hematopoietic progenitor cells (PBPC) modified with human aldehyde dehydrogenase class-3 gene (ALDH-3) and multidrug resistance gene 1 (MDR1) would increase chemotherapy resistance to 4-hydroperoxycyclophosphamide (4-HC) and -glycoprotein effluxed drugs.

METHODS:

A bicistronic retroviral vector G1Na-ALDH3-IRES-MDR1 cDNA was constructed and used to transfect the packaging cell lines PA317 by electroporation. CD34+ PBPC were isolated with a high-gradient magnetic cell sorting system (MACS), and then were transfected with supernatant of retrovirus containing human ALDH-3 and MDR1 cDNA. PCR, RT-PCR, Southern blot, Northern blot, FACS, and MTT assay were used to evaluate the transfection and expression of the transgene in target cells.

RESULTS:

The bicistronic retroviral vector construction was verified by PCR and restriction endonuclease analysis. Dual drug resistance genes were integrated into the genomic DNA of CD34+ PBPC and expressed efficiently. The efficiency of gene transfection in CD34+ PBPC was tested to be 18 % on colonies. Nested PCR and Neor rescue assay indicated that no helper virus was present in this system. Compared with the untransduced cells, transgene recipient cells conferred 4.5-fold resistance to 4-HC, 6.6-fold and 7.8-fold resistance to P-glycoprotein effluxed drug, vincristine and daunorubicin, respectively.

CONCLUSION:

Efficient transduction of two different types of drug resistance genes into human peripheral blood hematopoietic progenitor cells and the co-expression may decrease cumulative myelosuppression of combination chemotherapy.

PMID:
11749781
[Indexed for MEDLINE]
Free full text

Supplemental Content

Full text links

Icon for Acta Pharmacologica Sinica
Loading ...
Support Center