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Related Experiment Videos

Genetic changes in methotrexate-resistant mosquito cells.

F A Shotkoski1, A M Fallon

  • 1Department of Entomology, University of Minnesota, St. Paul 55108.

Archives of Insect Biochemistry and Physiology
|January 1, 1990
PubMed
Summary
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Recovery of cDNAs encoding ribosomal proteins S9 and L26 from Aedes albopictus mosquito cells and identification of their homologs in the malaria vector, Anopheles gambiae.

Archives of insect biochemistry and physiology·2005

Researchers developed methotrexate (mtx)-resistant Aedes albopictus mosquito cells. These cells showed significant mtx resistance due to dihydrofolate reductase (DHFR) gene amplification and chromosomal changes.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Methotrexate (mtx) is a chemotherapy drug that inhibits dihydrofolate reductase (DHFR).
  • Drug resistance in cancer cells often involves gene amplification and altered protein expression.
  • Aedes albopictus cells provide a model for studying drug resistance mechanisms.

Purpose of the Study:

  • To generate and characterize methotrexate-resistant variants of Aedes albopictus cells.
  • To investigate the molecular mechanisms underlying methotrexate resistance in these cells.

Main Methods:

  • Stepwise selection procedure to induce methotrexate resistance.
  • Growth assays to quantify resistance levels.
  • Gene amplification analysis (mRNA and DNA enrichment).

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  • Karyotype analysis to study chromosomal changes.

    • Main Results:

    • Generated Aedes albopictus cell variants 270- to 3,000-fold more resistant to methotrexate.
    • Observed amplification of the dihydrofolate reductase (DHFR) gene.
    • Confirmed DHFR gene amplification through increased DHFR protein and mRNA levels.
    • Identified chromosomal abnormalities including tetraploidy and rearrangements in resistant cells.

    Conclusions:

    • Dihydrofolate reductase (DHFR) gene amplification is a key mechanism for methotrexate resistance in Aedes albopictus cells.
    • Chromosomal alterations, including tetraploidy and rearrangements, are associated with high-level methotrexate resistance.
    • These resistant cell lines serve as valuable tools for studying drug resistance in insect cells.