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Basic Sciences

DNA-mediated Transfer and Cloning of a Human Multidrug-resistant Gene of Adriamycin-resistant Myelogenous Leukemia K562

Yoshikazu Sugimoto and Takashi Tsuruo
Yoshikazu Sugimoto
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Takashi Tsuruo
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DOI:  Published May 1987
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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.

Footnotes

  • ↵1 Supported by Grant-in-aid from the Ministry of Education, Science and Culture, Japan.

  • ↵2 To whom requests for reprints should be addressed.

  • Received September 11, 1986.
  • Revision received December 9, 1986.
  • Accepted February 13, 1987.
  • ©1987 American Association for Cancer Research.
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May 1987
Volume 47, Issue 10
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DNA-mediated Transfer and Cloning of a Human Multidrug-resistant Gene of Adriamycin-resistant Myelogenous Leukemia K562
Yoshikazu Sugimoto and Takashi Tsuruo
Cancer Res May 15 1987 (47) (10) 2620-2625;

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DNA-mediated Transfer and Cloning of a Human Multidrug-resistant Gene of Adriamycin-resistant Myelogenous Leukemia K562
Yoshikazu Sugimoto and Takashi Tsuruo
Cancer Res May 15 1987 (47) (10) 2620-2625;
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