Affiliations of authors: F. Pirnia, M. Breuleux, D. C. Betticher, M. M. Borner (Institute of Medical Oncology), M. A. Hotz (Department of Ear, Nose and Throat Surgery), Inselspital, Bern, Switzerland; M. Hochmeister (Department of Forensic Medicine), A. Marti (Department for Clinical Research), University of Bern; E. Schneider, Division of Molecular Medicine, Wadsworth Center, New York State Department of Health, Albany, NY; S. E. Bates, Medicine Branch, National Cancer Institute, Bethesda, MD.
Correspondence to: Markus M. Borner, M.D., Institute of Medical Oncology, Inselspital, 3010 Bern, Switzerland (e-mail: mborner{at}insel.ch).
The doxorubicin (ADR)-selected human breast cancer cell line (MCF-7/ADR-RES) has been a useful model for a multidrug-resistant subline in cancer research. However, a recent letter to the Journal has alerted the scientific community about the finding that MCF-7/ADR-RES is not a MCF-7-derived cell line. This report led to a change of nomenclature from MCF-7/ADR-RES to NCI/ADR-RES (1).
Another cell line (MCF-7 TH) has been independently selected by doxorubicin treatment at the National Cancer Institute (NCI) and demonstrated some of the same, interesting characteristics of NCI/ADR-RES, such as high levels of MDR-1 and P-glycoprotein expression and a mutated p53 tumor suppressor protein (2). Of interest, both cell lines exhibit the same 21-base-pair deletion beginning at exon 5 spanning codons 126133 (3). We used this cell line to analyze the effect of the multidrug-resistant phenotype and p53 on the efficacy of anticancer drugs with different mechanisms of action. Various anticancer drugs were more effective in the MCF-7 TH subline than in the parental MCF-7 line, despite a dysfunctional p53 and multidrug-resistant phenotype. These agents induced apoptosis in MCF-7 TH cells as demonstrated by the characteristic morphologic features. Because the activity of specific caspases might affect the propensity of a cell to undergo apoptosis, we examined expression level and cleavage pattern of different caspases in drug-treated MCF-7 TH and parental cells. The MCF-7 TH cell line strongly expressed caspase-3 (Fig. 1, A) and cisplatin treatment led to a significant increase of caspase-3-like activity as measured by use of the fluorogenic caspase-3-specific substrate Asp-Glu-Val-Asp-7-amino-4-trifluoromethyl coumarin (DEVD-afc) (Fig. 1
, B). The MCF-7 cell line has lost caspase-3 because of a 47-base-pair deletion within exon 3 of the CASP-3 gene (4). These findings thus suggest that MCF-7 TH also is not derived from MCF-7. This hypothesis was confirmed by DNA fingerprinting, which demonstrated that parental MCF-7 and MCF-7 TH cannot have the same donor, because they differ in all loci except the sex chromosome locus. The analyses were repeated with several different batches of parental MCF-7 and TH cells. Furthermore, MCF-7 parental cells from different sources were compared and all showed an identical allelic distribution profile of the short tandem repeat loci by DNA fingerprinting. The only cell line showing identity with MCF-7 TH by DNA fingerprinting was NCI/ADR-RES. Thus, MCF-7 TH, like NCI/ADR-RES, is not a MCF-7-related cell line.
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