Abstract
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.
Highlights
AND HISTORICAL PERSPECTIVE The development of chemotherapy drug-resistant cancer cell lines is a long established approach for investigating the mechanisms of cytotoxicity and resistance to chemotherapy agents
This induced 2500-fold greater resistance to the drug than that observed in the parental cells. These resistant cell lines were cross resistant to other chemotherapy drugs such as vinblastine and daunorubicin
We found that the 50% inhibitory concentration (IC50) concentrations of temozolomide were in the high micromolar range in melanoma cell lines
Summary
AND HISTORICAL PERSPECTIVE The development of chemotherapy drug-resistant cancer cell lines is a long established approach for investigating the mechanisms of cytotoxicity and resistance to chemotherapy agents. H69CIS200 and H69OX400 cisplatin and oxaliplatin-resistant cells were 1.5–2-fold resistant to platinums for 5–6 weeks in drug-free culture and the resistance phenotype faded over the 6–8 weeks in culture [20] This technique is often used with resistant models regardless of selection strategy to ensure consistency, so that cells within a limited range of passage numbers are used for all experiments. It should be noted, that different research groups can have different sub clones of a parent cell line and this can be a factor for the differences in the resistant models produced These examples demonstrate that the same cell line, treated with the same chemotherapy agent leads to the development of a heterogeneous range of drug-resistant models. If parallel models of the same treatment are produced the heterogeneity between drug-resistant cell lines should be examined with interest rather than dismissed as a non-reproducible experiment
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