In vitro Development of Chemotherapy and Targeted Therapy Drug-Resistant Cancer Cell Lines: A Practical Guide with Case Studies.

Article OPEN
BUSSCHOTS, STEVEN (2014)
  • Publisher: Frontiers Media S.A.
  • Journal: volume 4 (issn: 2234-943X, eissn: 2234-943X)
  • Related identifiers: doi: 10.3389/fonc.2014.00040, pmc: PMC3944788
  • Subject: Methods Article | cell lines | drug-resistant cell line | drug-resistance | selection strategy | cancer | chemotherapy | Oncology

PUBLISHED The development of a drug-resistant cell line can take from 3 to 18 months. However, little is published on the methodology of this development process. This article will discuss key decisions to be made prior to starting resistant cell line development; the choice of parent cell line, dose of selecting agent, treatment interval, and optimizing the dose of drug for the parent cell line. Clinically relevant drug-resistant cell lines are developed by mimicking the conditions cancer patients experience during chemotherapy and cell lines display between two- and eight-fold resistance compared to their parental cell line. Doses of drug administered are low, and a pulsed treatment strategy is often used where the cells recover in drug-free media. High-level laboratory models are developed with the aim of understanding potential mechanisms of resistance to chemotherapy agents. Doses of drug are higher and escalated over time. It is common to have difficulty developing stable clinically relevant drug-resistant cell lines. A comparative selection strategy of multiple cell lines or multiple chemotherapeutic agents mitigates this risk and gives insight into which agents or type of cell line develops resistance easily. Successful selection strategies from our research are presented. Pulsed-selection produced platinum or taxane-resistant large cell lung cancer (H1299 and H460) and temozolomide-resistant melanoma (Malme-3M and HT144) cell lines. Continuous selection produced a lapatinib-resistant breast cancer cell line (HCC1954). Techniques for maintaining drug-resistant cell lines are outlined including; maintaining cells with chemotherapy, pulse treating with chemotherapy, or returning to master drug-resistant stocks. The heterogeneity of drug-resistant models produced from the same parent cell line with the same chemotherapy agent is explored with reference to P-glycoprotein. Heterogeneity in drug-resistant cell lines reflects the heterogeneity that can occur in clinical drug resistance.
  • References (97)
    97 references, page 1 of 10

    1. Biedler JL, Riehm H. Cellular resistance to actinomycin D in Chinese hamster cells in vitro: cross-resistance, radioautographic, and cytogenetic studies. Cancer Res (1970) 30(4):1174-84.

    2. Roberts D, Wodinsky I, Hall TC. Studies on folic reductase. Cancer Res (1965) 25(11 Pt 1):1899-903.

    3. Burchenal JH, Robinson E, Johnston SF, Kushida MN. The induction of resistance to 4-amino-N10-methyl-pteroylglutamic acid in a strain of transmitted mouse leukemia. Science (1950) 111(2875):116-7. doi:10.1126/science.111. 2875.116

    4. Kessel D, Botterill V, Wodinsky I. Uptake and retention of daunomycin by mouse leukemic cells as factors in drug response. Cancer Res (1968) 28(5):938-41.

    5. Law LW. Resistance in leukemic cells to a guanine analog, 8-azaguanine. Proc Soc Exp Biol Med (1951) 78(2):499-502. doi:10.3181/00379727-78-19118

    6. Martin A, Clynes M. Comparison of 5 microplate colorimetric assays for in vitro cytotoxicity testing and cell proliferation assays. Cytotechnology (1993) 11(1):49-58. doi:10.1007/BF00749057

    7. Franken NAP, Rodermond HM, Stap J, Haveman J, van Bree C. Clonogenic assay of cells in vitro. Nat Protoc (2006) 1(5):2315-9. doi:10.1038/nprot.2006.339

    8. Kawai H, Kiura K, Tabata M,Yoshino T, Takata I, Hiraki A, et al. Characterization of non-small-cell lung cancer cell lines established before and after chemotherapy. Lung Cancer (2002) 35(3):305-14. doi:10.1016/S0169-5002(01)00430-5

    9. Kuroda H, Sugimoto T, Ueda K, Tsuchida S, Horii Y, Inazawa J, et al. Different drug sensitivity in two neuroblastoma cell lines established from the same patient before and after chemotherapy. Int J Cancer (1991) 47(5):732-7. doi:10.1002/ijc.2910470518

    10. Hida T, Ueda R, Takahashi T, Watanabe H, Kato T, Suyama M, et al. Chemosensitivity and radiosensitivity of small cell lung cancer cell lines studied by a newly developed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) hybrid assay. Cancer Res (1989) 49(17):4785-90.

  • Related Research Results (1)
  • Similar Research Results (1)
  • Metrics
    No metrics available
Share - Bookmark