Apoptotic response to camptothecin and 7‐hydroxystaurosporine (UCN‐01) in the 8 human breast cancer cell lines of the NCI anticancer drug screen: Multifactorial relationships with topoisomerase i, protein kinase C, Bcl‐2, p53, MDM‐2 and caspase pathways

Derivatives of camptothecins, topoisomerase I inhibitors and 7‐hydroxystaurosporine (UCN‐01), a protein kinase C (PKC) inhibitor and cell cycle checkpoint abrogator, are promising anticancer drugs. We characterized the apoptotic response to camptothecin and UCN‐01 for the 8 human breast carcinoma cell lines (MCF‐7, MCF‐7/ADR, T47D, HS578T, BT549, MDA‐N, MDA MB231, MDA435) from the National Cancer Institute (NCI) Anticancer Drug Screen. MCF‐7 and T47D cells exhibited marked resistance to apoptosis, whereas MCF‐7/ADR (NCI/ADR‐RES) and HS578T cells exhibited the most pronounced apoptotic response. Apoptotic response was not correlated with growth inhibition measured by sulforhodamine B (SRB) assay, indicating that apoptosis is not the only mechanism of drug‐induced cell death. Measurements of topoisomerase I levels and cleavage complexes and of PKC isoforms demonstrated that primary target inhibition was not correlated with apoptotic response. Several key apoptotic pathways were evaluated. Only MCF‐7 cells had wild‐type p53, indicating that p53 is not required for drug‐induced apoptosis. MCF‐7 cells also showed the highest MDM‐2 expression (along with T47D cells, which were also resistant to apoptosis). Bcl‐2, Mcl‐1 and caspases 2 and 3 protein levels varied widely, whereas Bax expression was comparable among cell lines. Interestingly, Bcl‐2, Mcl‐1 and Bcl‐XL cumulative expressions were inversely correlated with apoptotic response. Our results provide a comparative molecular characterization for the breast cancer cell lines of the NCI Anticancer Drug Screen and demonstrate the diversity of cellular responses to drugs (apoptosis vs. cell cycle arrest) and the importance of multifactorial analyses for modulating/predicting the apoptotic response to chemotherapy. Int. J. Cancer 82:396–404, 1999. Published 1999 Wiley‐Liss, Inc.

[1]  S. Korsmeyer,et al.  Cell Death in Development , 1999, Cell.

[2]  Alan G. Porter,et al.  Caspase-3 Is Required for DNA Fragmentation and Morphological Changes Associated with Apoptosis* , 1998, The Journal of Biological Chemistry.

[3]  E. Schmitt,et al.  Bcl-xL modulates apoptosis induced by anticancer drugs and delays DEVDase and DNA fragmentation-promoting activities. , 1998, Experimental cell research.

[4]  John Calvin Reed,et al.  BCL-X expression in multiple myeloma: possible indicator of chemoresistance. , 1998, Cancer research.

[5]  J N Weinstein,et al.  Characterization of the p53 tumor suppressor pathway in cell lines of the National Cancer Institute anticancer drug screen and correlations with the growth-inhibitory potency of 123 anticancer agents. , 1997, Cancer research.

[6]  K. Kohn,et al.  Abrogation of an S-phase checkpoint and potentiation of camptothecin cytotoxicity by 7-hydroxystaurosporine (UCN-01) in human cancer cell lines, possibly influenced by p53 function. , 1997, Cancer research.

[7]  M. Shimizu,et al.  G1 phase accumulation induced by UCN-01 is associated with dephosphorylation of Rb and CDK2 proteins as well as induction of CDK inhibitor p21/Cip1/WAF1/Sdi1 in p53-mutated human epidermoid carcinoma A431 cells. , 1997, Cancer research.

[8]  B. Ogretmen,et al.  Expression of the mutated p53 tumor suppressor protein and its molecular and biochemical characterization in multidrug resistant MCF-7/Adr human breast cancer cells , 1997, Oncogene.

[9]  Y. Pommier,et al.  Cellular Resistance to Camptothecins , 1996, Annals of the New York Academy of Sciences.

[10]  E. Sausville,et al.  UCN-01: a potent abrogator of G2 checkpoint function in cancer cells with disrupted p53. , 1996, Journal of the National Cancer Institute.

[11]  T Takahashi,et al.  Overexpression of bax sensitizes human breast cancer MCF‐7 cells to radiation‐induced apoptosis , 1996, International journal of cancer.

[12]  A. Eastman,et al.  Enhancement of cisplatin-induced cytotoxicity by 7-hydroxystaurosporine (UCN-01), a new G2-checkpoint inhibitor. , 1996, Clinical cancer research : an official journal of the American Association for Cancer Research.

[13]  W. El-Deiry,et al.  Apoptotic death of tumor cells correlates with chemosensitivity, independent of p53 or bcl-2. , 1996, Clinical cancer research : an official journal of the American Association for Cancer Research.

[14]  Y. Pommier.  Eukaryotic DNA topoisomerase I: genome gatekeeper and its intruders, camptothecins. , 1996, Seminars in oncology.

[15]  Mcvie Jg.  Current areas of treatment. , 1996 .

[16]  Y. Pommier,et al.  The role of cell cycle regulation and apoptosis triggering in determining the sensitivity of leukemic cells to topoisomerase I and II inhibitors. , 1995, Leukemia.

[17]  Y. Pommier,et al.  Topoisomerase I-related parameters and camptothecin activity in the colon carcinoma cell lines from the National Cancer Institute anticancer screen. , 1995, Cancer research.

[18]  D. Housman,et al.  p53 status and the efficacy of cancer therapy in vivo. , 1994, Science.

[19]  Y. Pommier,et al.  bcl-2 protein inhibits etoposide-induced apoptosis through its effects on events subsequent to topoisomerase II-induced DNA strand breaks and their repair. , 1993, Cancer research.

[20]  E. Sausville,et al.  Cell cycle arrest and growth inhibition by the protein kinase antagonist UCN-01 in human breast carcinoma cells. , 1993, Cancer research.

[21]  S. Korsmeyer,et al.  Constitutive expression of human Bcl-2 modulates nitrogen mustard and camptothecin induced apoptosis. , 1993, Cancer research.

[22]  John Calvin Reed,et al.  bcl-2 gene transfer increases relative resistance of S49.1 and WEHI7.2 lymphoid cells to cell death and DNA fragmentation induced by glucocorticoids and multiple chemotherapeutic drugs. , 1992, Cancer research.

[23]  Y. Cheng,et al.  Characterization of camptothecin-resistant Chinese hamster lung cells. , 1992, Biochemical pharmacology.

[24]  S. Sukumar,et al.  Mutations in p53 as potential molecular markers for human breast cancer. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Bartek,et al.  Genetic and immunochemical analysis of mutant p53 in human breast cancer cell lines. , 1990, Oncogene.

[26]  P. Pantazis,et al.  The camptothecins : from discovery to the patient , 1996 .