DNA-mediated transfer and cloning of a human multidrug-resistant gene of Adriamycin-resistant myelogenous leukemia K562

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Abstract

We have successfully transferred and cloned a fragment of a human multidrug-resistant gene by using DNA-mediated gene transfer. Macromolecular DNA of human multidrug-resistant K562 cells was transfected to drug-sensitive mouse Ltk- cells to obtain a drug-resistant transfectant with a human resistant gene. Both primary and secondary transfectants showed similar patterns of cross-resistance to Adriamycin and vincristine. The mechanism of drug resistance of the transfectants was attributed to decreased retention of the drug. Three secondary transfectants obtained independently contained common Alu-containing EcoRI fragments 15, 6.5, 3.7, 2.6, and 1.9 kilobases long. The 2.6-kilobase EcoRI fragment was cloned from a λ phage genomic library made from DNA of a secondary transfectant. The 2.6-kilobase fragment was detected in the primary and secondary transfectants but not in the parental Ltk-, Adriamycin-resistant Ltk-, and Adriamycin-resistant P388 cells. This sequence was found to be amplified in several multidrug-resistant cell lines such as Adriamycin-resistant ovarian carcinoma A2780 and colchicine-resistant KB carcinoma cells. The 2.6-kilobase fragment hybridized with a 4.5-kilobase mRNA which is overexpressed in the Adriamycin-resistant K562 cells and the Adriamycin-resistant A2780 cells but not detected in the parental K562 cells. The gene transferred and cloned in this study seems to be related to the P-glycoprotein gene as judged from the size of mRNA and its overexpression in some of the multidrug-resistant cell lines where P-glycoprotein was found to be highly expressed.

Original languageEnglish
Pages (from-to)2620-2625
Number of pages6
JournalCancer Research
Volume47
Issue number10
Publication statusPublished - 1987
Externally publishedYes

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Myeloid Leukemia
Doxorubicin
Organism Cloning
K562 Cells
DNA
Genes
P-Glycoprotein
Pharmaceutical Preparations
Carcinoma
KB Cells
Cell Line
Messenger RNA
Genomic Library
Colchicine
Vincristine
Drug Resistance
Bacteriophages

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

DNA-mediated transfer and cloning of a human multidrug-resistant gene of Adriamycin-resistant myelogenous leukemia K562. / Sugimoto, Yoshikazu; Tsuruo, T.

In: Cancer Research, Vol. 47, No. 10, 1987, p. 2620-2625.

Research output: Contribution to journalArticle

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