Cell Cycle Phase-Specific Drug Resistance as an Escape Mechanism of Melanoma Cells.

The tumor microenvironment is characterized by cancer cell subpopulations with heterogeneous cell cycle profiles. For example, hypoxic tumor zones contain clusters of cancer cells that arrest in G1 phase. It is conceivable that neoplastic cells exhibit differential drug sensitivity based on their residence in specific cell cycle phases. In this study, we used two-dimensional and organotypic melanoma culture models in combination with fluorescent cell cycle indicators to investigate the effects of cell cycle phases on clinically used drugs. We demonstrate that G1-arrested melanoma cells, irrespective of the underlying cause mediating G1 arrest, are resistant to apoptosis induced by the proteasome inhibitor bortezomib or the alkylating agent temozolomide. In contrast, G1-arrested cells were more sensitive to mitogen-activated protein kinase pathway inhibitor-induced cell death. Of clinical relevance, pretreatment of melanoma cells with a mitogen-activated protein kinase pathway inhibitor, which induced G1 arrest, resulted in resistance to temozolomide or bortezomib. On the other hand, pretreatment with temozolomide, which induced G2 arrest, did not result in resistance to mitogen-activated protein kinase pathway inhibitors. In summary, we established a model to study the effects of the cell cycle on drug sensitivity. Cell cycle phase-specific drug resistance is an escape mechanism of melanoma cells that has implications on the choice and timing of drug combination therapies.

[1]  A. Ribas,et al.  New drug targets in metastatic melanoma , 2014, The Journal of pathology.

[2]  G. Schwartz,et al.  Sequential dependent enhancement of caspase activation and apoptosis by flavopiridol on paclitaxel-treated human gastric and breast cancer cells. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[3]  Kam Y. J. Zhang,et al.  Discovery of a selective inhibitor of oncogenic B-Raf kinase with potent antimelanoma activity , 2008, Proceedings of the National Academy of Sciences.

[4]  Timothy J. Mitchison,et al.  The proliferation rate paradox in antimitotic chemotherapy , 2012, Molecular biology of the cell.

[5]  N. Haass,et al.  Targeting the intrinsic apoptosis pathway as a strategy for melanoma therapy , 2014, Pigment cell & melanoma research.

[6]  Tae Won Kim,et al.  Bortezomib induces G2-M arrest in human colon cancer cells through ROS-inducible phosphorylation of ATM-CHK1. , 2012, International journal of oncology.

[7]  Ann Richmond,et al.  Augmenting Chemosensitivity of Malignant Melanoma Tumors via Proteasome Inhibition , 2004, Cancer Research.

[8]  M. Herlyn,et al.  The Mitogen-Activated Protein/Extracellular Signal-Regulated Kinase Kinase Inhibitor AZD6244 (ARRY-142886) Induces Growth Arrest in Melanoma Cells and Tumor Regression When Combined with Docetaxel , 2008, Clinical Cancer Research.

[9]  N. Saijo,et al.  Bortezomib potentially inhibits cellular growth of vascular endothelial cells through suppression of G2/M transition , 2010, Cancer science.

[10]  M. Amiot,et al.  A pivotal role for Mcl-1 in Bortezomib-induced apoptosis , 2008, Oncogene.

[11]  M. Herlyn,et al.  In vitro three-dimensional tumor microenvironment models for anticancer drug discovery , 2008, Expert opinion on drug discovery.

[12]  Loredana Spoerri,et al.  Real-Time Cell Cycle Imaging in a 3D Cell Culture Model of Melanoma. , 2017, Methods in molecular biology.

[13]  Aoife M Shannon,et al.  Tumour hypoxia, chemotherapeutic resistance and hypoxia-related therapies. , 2003, Cancer treatment reviews.

[14]  N. Haass,et al.  Evaluation of Cell Cycle Arrest in Estrogen Responsive MCF-7 Breast Cancer Cells: Pitfalls of the MTS Assay , 2011, PloS one.

[15]  Yuchun Luo,et al.  ABT-737 synergizes with Bortezomib to kill melanoma cells , 2011, Biology Open.

[16]  N. Haass,et al.  Melanoma's connections to the tumour microenvironment. , 2013, Pathology.

[17]  B. Kaina,et al.  Apoptosis triggered by DNA damage O6-methylguanine in human lymphocytes requires DNA replication and is mediated by p53 and Fas/CD95/Apo-1 , 2004, Oncogene.

[18]  M. Belvin,et al.  RAF inhibitors prime wild-type RAF to activate the MAPK pathway and enhance growth , 2010, Nature.

[19]  T. Burke,et al.  The CDK4/6 Inhibitor LY2835219 Overcomes Vemurafenib Resistance Resulting from MAPK Reactivation and Cyclin D1 Upregulation , 2013, Molecular Cancer Therapeutics.

[20]  N. Haass,et al.  Induction of endoplasmic reticulum stress as a strategy for melanoma therapy: is there a future? , 2014, Melanoma management.

[21]  P. Hersey,et al.  Tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis of human melanoma is regulated by smac/DIABLO release from mitochondria. , 2001, Cancer research.

[22]  Eric Brown,et al.  Up-regulated expression of zonula occludens protein-1 in human melanoma associates with N-cadherin and contributes to invasion and adhesion. , 2005, The American journal of pathology.

[23]  J. Utikal,et al.  Dabrafenib and trametinib versus dabrafenib and placebo for Val600 BRAF-mutant melanoma: a multicentre, double-blind, phase 3 randomised controlled trial , 2015, The Lancet.

[24]  N. Haass.  Dynamic tumor heterogeneity in melanoma therapy: how do we address this in a novel model system? , 2015, Melanoma management.

[25]  S. Nikfar,et al.  Efficacy and side effects of dacarbazine in comparison with temozolomide in the treatment of malignant melanoma: a meta-analysis consisting of 1314 patients , 2013, Melanoma research.

[26]  N. Haass,et al.  Real‐time cell cycle imaging during melanoma growth, invasion, and drug response , 2014, Pigment cell & melanoma research.

[27]  S. Lowe,et al.  Differential regulation of noxa in normal melanocytes and melanoma cells by proteasome inhibition: therapeutic implications. , 2005, Cancer research.

[28]  K. Flaherty,et al.  Combined BRAF (Dabrafenib) and MEK inhibition (Trametinib) in patients with BRAFV600-mutant melanoma experiencing progression with single-agent BRAF inhibitor. , 2014, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[29]  I. Barasoain,et al.  Taxanes: microtubule and centrosome targets, and cell cycle dependent mechanisms of action. , 2003, Current cancer drug targets.

[30]  D. Kuhn,et al.  Proteasome Inhibitors in Cancer Therapy: Lessons from the First Decade , 2008, Clinical Cancer Research.

[31]  Victor G Prieto,et al.  Integrated Molecular and Clinical Analysis of AKT Activation in Metastatic Melanoma , 2009, Clinical Cancer Research.

[32]  P. Elliott,et al.  Mechanisms of proteasome inhibitor PS-341-induced G(2)-M-phase arrest and apoptosis in human non-small cell lung cancer cell lines. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[33]  H. Drexler.  Activation of the cell death program by inhibition of proteasome function. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Simeon Santourlidis,et al.  Bortezomib/proteasome inhibitor triggers both apoptosis and autophagy-dependent pathways in melanoma cells. , 2013, Cellular signalling.

[35]  Keith T Flaherty,et al.  Multiple signaling pathways must be targeted to overcome drug resistance in cell lines derived from melanoma metastases , 2006, Molecular Cancer Therapeutics.

[36]  M. Soengas,et al.  Therapeutic window for melanoma treatment provided by selective effects of the proteasome on Bcl-2 proteins , 2007, Cell Death and Differentiation.

[37]  Z. Jehan,et al.  Bortezomib stabilizes mitotic cyclins and prevents cell cycle progression via inhibition of UBE2C in colorectal carcinoma. , 2011, The American journal of pathology.

[38]  K. Black,et al.  Proteasome inhibitor PS-341 causes cell growth arrest and apoptosis in human glioblastoma multiforme (GBM) , 2005, Oncogene.

[39]  P. Hersey,et al.  The BH3-mimetic ABT-737 sensitizes human melanoma cells to apoptosis induced by selective BRAF inhibitors but does not reverse acquired resistance. , 2013, Carcinogenesis.

[40]  N. Erin,et al.  Differential sensitivity of breast cancer and melanoma cells to proteasome inhibitor Velcade. , 2008, International journal of molecular medicine.

[41]  K. Flaherty,et al.  Combined BRAF and MEK inhibition in melanoma with BRAF V600 mutations. , 2012, The New England journal of medicine.

[42]  M. Herlyn,et al.  PLX4032, a potent inhibitor of the B‐Raf V600E oncogene, selectively inhibits V600E‐positive melanomas , 2010, Pigment cell & melanoma research.

[43]  M. Herlyn,et al.  Melanocytes: From Morphology to Application , 2009, Skin Pharmacology and Physiology.

[44]  K. Flaherty,et al.  An organometallic protein kinase inhibitor pharmacologically activates p53 and induces apoptosis in human melanoma cells. , 2007, Cancer research.

[45]  Farzana Ahmed,et al.  Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids. , 2015, Journal of visualized experiments : JoVE.

[46]  K. Flaherty,et al.  Ki67 expression levels are a better marker of reduced melanoma growth following MEK inhibitor treatment than phospho-ERK levels , 2007, British Journal of Cancer.

[47]  R. Flockhart,et al.  Combining the Endoplasmic Reticulum Stress–Inducing Agents Bortezomib and Fenretinide as a Novel Therapeutic Strategy for Metastatic Melanoma , 2009, Clinical Cancer Research.

[48]  S. Haferkamp,et al.  Vemurafenib induces senescence features in melanoma cells. , 2013, The Journal of investigative dermatology.

[49]  A. Giobbie-Hurder,et al.  A Phase I Trial of Bortezomib and Sorafenib in Advanced Malignant Melanoma. , 2015, The oncologist.

[50]  R. Scott,et al.  Molecular and Cellular Pathobiology Nucleotide Excision Repair Gene Expression after Cisplatin Treatment in Melanoma , 2022 .

[51]  Nikolas K. Haass,et al.  Modeling Melanoma In Vitro and In Vivo , 2013, Healthcare.

[52]  J A Wohlschlegel,et al.  Inhibition of eukaryotic DNA replication by geminin binding to Cdt1. , 2000, Science.

[53]  D. Schadendorf,et al.  Metastatic potential of melanomas defined by specific gene expression profiles with no BRAF signature. , 2006, Pigment cell research.

[54]  M. Christmann,et al.  B-Raf inhibitor vemurafenib in combination with temozolomide and fotemustine in the killing response of malignant melanoma cells , 2014, Oncotarget.

[55]  S. Kridel,et al.  Preferential cytotoxicity of bortezomib toward hypoxic tumor cells via overactivation of endoplasmic reticulum stress pathways. , 2008, Cancer research.

[56]  B. Kaina,et al.  Contribution of ATM and ATR to the Resistance of Glioblastoma and Malignant Melanoma Cells to the Methylating Anticancer Drug Temozolomide , 2013, Molecular Cancer Therapeutics.

[57]  Nizar M. Mhaidat,et al.  Temozolomide induces senescence but not apoptosis in human melanoma cells , 2007, British Journal of Cancer.

[58]  K. L. Donaldson,et al.  Cytotoxicity of the anticancer agents cisplatin and taxol during cell proliferation and the cell cycle , 1994, International journal of cancer.

[59]  H P Soyer,et al.  A stress-induced early innate response causes multidrug tolerance in melanoma , 2014, Oncogene.

[60]  Atsushi Miyawaki,et al.  Visualizing Spatiotemporal Dynamics of Multicellular Cell-Cycle Progression , 2008, Cell.

[61]  M. Scheffner,et al.  Activation of the Cell Death Program by Nitric Oxide Involves Inhibition of the Proteasome* , 1999, The Journal of Biological Chemistry.

[62]  M. Hendrix,et al.  Proteasome inhibitors trigger NOXA-mediated apoptosis in melanoma and myeloma cells. , 2005, Cancer research.

[63]  A. Weeraratna,et al.  Hypoxia induces phenotypic plasticity and therapy resistance in melanoma via the tyrosine kinase receptors ROR1 and ROR2. , 2013, Cancer discovery.

[64]  G. McArthur,et al.  Cell cycle control as a promising target in melanoma , 2015, Current opinion in oncology.

[65]  G. McArthur.  Combination Therapies to Inhibit the RAF/MEK/ERK Pathway in Melanoma: We are not Done Yet , 2015, Front. Oncol..

[66]  W. Hahn,et al.  Prolonged early G(1) arrest by selective CDK4/CDK6 inhibition sensitizes myeloma cells to cytotoxic killing through cell cycle-coupled loss of IRF4. , 2012, Blood.

[67]  J. Sosman,et al.  A Phase I Trial of Bortezomib with Temozolomide in Patients with Advanced Melanoma: Toxicities, Antitumor Effects, and Modulation of Therapeutic Targets , 2009, Clinical Cancer Research.

[68]  P. Hersey,et al.  Modulation of NOXA and MCL-1 as a Strategy for Sensitizing Melanoma Cells to the BH3-Mimetic ABT-737 , 2011, Clinical Cancer Research.