Retroviral coexpression of two different types of drug resistance genes to protect normal cells from combination chemotherapy

Mutsumi Suzuki, Yoshikazu Sugimoto, Satomi Tsukahara, Etsuko Okochi, Michael M. Gottesman, Takashi Tsuruo

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Drug resistance genes can protect normal hematopoietic cells from the toxicity of anticancer agents. Because chemotherapeutic agents are often used in combination in current clinical protocols, coexpression of two different drug resistance genes should be useful in protecting normal bone marrow cells from the hematotoxicities caused by combination chemotherapy. In this study, we have combined the human multidrug resistance gene (MDR1) and human O6-methylguanine DNA methyltransferase (MGMT) gene as drug resistance genes. For the coexpression of two drug resistance genes, we have constructed two bicistronic retrovirus vectors. One vector is Ha-MDR-IRES-MGMT, in which translation of the MDR1 cDNA is cap-dependent and MGMT translation is dependent on an internal ribosome entry site (IRES). The other is Ha-MGMT-IRES-MDR, which has cap-dependent MGMT translation and IRES-dependent MDR1 translation, MGMT-negative HeLa derivative (MR) cells transduced with these retroviruses showed resistance to vincristine (from MDR1) and 1-(4-amino-2-methyl-5- -pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU; from MGMT). Cells transduced with Ha-MDR- IRES-MGMT showed higher resistance to vincristine and lower resistance to ACNU than those transduced with Ha-MGMT-IRES-MDR. In any case, the resistance levels of cells transduced with either vector were high enough to select transduced cells with vincristine or ACNU. The expression levels of P-glycoprotein or MGMT in the transduced cells determined by FAGS and Western blot analysis correlated well with the extent of resistance to vincristine and ACNU, respectively. All of the MGMT-transduced cells expressed higher amounts of MGMT than the MGMT-expressing parental cell line HeLa S3. Murine bone marrow cells transduced with Ha-MDR-IRES-MGMT and selected with vincristine also showed simultaneous resistance to vincristine and ACNU. These results suggest that bicistronic retroviral vectors allow the functional coexpression of two different types of drug resistance genes. This strategy could be applicable to any combination of drug resistance genes.

Original languageEnglish
Pages (from-to)947-954
Number of pages8
JournalClinical Cancer Research
Volume3
Issue number6
Publication statusPublished - 1997 Jun
Externally publishedYes

Fingerprint

Methyltransferases
Combination Drug Therapy
Drug Resistance
DNA
Nimustine
Vincristine
Genes
Retroviridae
Bone Marrow Cells
MDR Genes
P-Glycoprotein
Clinical Protocols
Internal Ribosome Entry Sites
Antineoplastic Agents
Complementary DNA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Retroviral coexpression of two different types of drug resistance genes to protect normal cells from combination chemotherapy. / Suzuki, Mutsumi; Sugimoto, Yoshikazu; Tsukahara, Satomi; Okochi, Etsuko; Gottesman, Michael M.; Tsuruo, Takashi.

In: Clinical Cancer Research, Vol. 3, No. 6, 06.1997, p. 947-954.

Research output: Contribution to journalArticle

Suzuki, Mutsumi ; Sugimoto, Yoshikazu ; Tsukahara, Satomi ; Okochi, Etsuko ; Gottesman, Michael M. ; Tsuruo, Takashi. / Retroviral coexpression of two different types of drug resistance genes to protect normal cells from combination chemotherapy. In: Clinical Cancer Research. 1997 ; Vol. 3, No. 6. pp. 947-954.
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