Inhibition of CDK4 sensitizes multidrug resistant ovarian cancer cells to paclitaxel by increasing apoptosiss
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Overexpression of cyclin-dependent kinase (CDK) 4 has been observed in a variety of cancers and has been found to contribute to tumor cell growth and proliferation. However, the effect of inhibition of CDK4 in ovarian cancer is unknown. We investigated the therapeutic effect of the CDK4 inhibitor palbociclib in combination with paclitaxel in ovarian cancer cells.
Cell viabilities were determined by MTT assay after exposure to different dosages of palbociclib and/or paclitaxel. Western blot, immunofluorescence, and Calcein AM assays were conducted to determine the mechanisms underlying the cytotoxic effects of palbociclib in combination with paclitaxel. CDK4 siRNA was used to validate the outcome of targeting CDK4 by palbociclib in ovarian cancer cells.
We found that combinations of palbociclib and paclitaxel significantly enhanced drug sensitivity in both Rb-positive (SKOV3TR) and Rb-negative (OVCAR8TR) ovarian cancer-derived cells. When combined with paclitaxel, palbociclib induced apoptosis in both SKOV3TR and OVCAR8TR cells. We also found that palbociclib inhibited the activity of P-glycoprotein (Pgp), and that siRNA-mediated CDK4 knockdown sensitized multidrug resistant (MDR) SKOV3TR and OVCAR8TR cells to paclitaxel.
Inhibition of CDK4 by palbociclib can enhance paclitaxel sensitivity in both Rb-positive and Rb-negative MDR ovarian cancer cells by increasing apoptosis. CDK4 may serve as a promising target in the treatment of ovarian cancer.
KeywordsOvarian cancer CDK4 Palbociclib Rb Paclitaxel Apoptosis MDR
This work was supported in part by grants from the Gattegno and Wechsler funds. Dr. Duan is supported, in part, through a grant from the Sarcoma Foundation of America (SFA), a pilot grant from Sarcoma SPORE/NIH, and a grant from the National Cancer Institute (NCI)/National Institutes of Health (NIH), UO1, CA 151452.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
- 3.M.A. Bookman, M.F. Brady, W.P. McGuire, P.G. Harper, D.S. Alberts, M. Friedlander, N. Colombo, J.M. Fowler, P.A. Argenta, K. De Geest, D.G. Mutch, R.A. Burger, A.M. Swart, E.L. Trimble, C. Accario-Winslow, L.M. Roth, Evaluation of new platinum-based treatment regimens in advanced-stage ovarian cancer: a phase III trial of the gynecologic cancer intergroup. J. Clin. Oncol. 27, 1419–1425 (2009)CrossRefPubMedPubMedCentralGoogle Scholar
- 4.S. Vaughan, J.I. Coward, R.C. Bast Jr., A. Berchuck, J.S. Berek, J.D. Brenton, G. Coukos, C.C. Crum, R. Drapkin, D. Etemadmoghadam, M. Friedlander, H. Gabra, S.B. Kaye, C.J. Lord, E. Lengyel, D.A. Levine, I.A. McNeish, U. Menon, G.B. Mills, K.P. Nephew, A.M. Oza, A.K. Sood, E.A. Stronach, H. Walczak, D.D. Bowtell, F.R. Balkwill, Rethinking ovarian cancer: recommendations for improving outcomes. Nat. Rev. Cancer 11, 719–725 (2011)CrossRefPubMedPubMedCentralGoogle Scholar
- 5.B.Y. Karlan, A.M. Oza, G.E. Richardson, D.M. Provencher, V.L. Hansen, M. Buck, S.K. Chambers, P. Ghatage, C.H. Pippitt Jr., J.V. Brown 3rd, A. Covens, R.V. Nagarkar, M. Davy, C.A. Leath 3rd, H. Nguyen, D.E. Stepan, D.M. Weinreich, M. Tassoudji, Y.N. Sun, I.B. Vergote, Randomized, double-blind, placebo-controlled phase II study of AMG 386 combined with weekly paclitaxel in patients with recurrent ovarian cancer. J. Clin. Oncol. 30, 362–371 (2012)CrossRefPubMedGoogle Scholar
- 10.Y. Liao, Y. Feng, J. Shen, F.J. Hornicek and Z. Duan, The roles and therapeutic potential of cyclin-dependent kinases (CDKs) in sarcoma. Cancer Metastasis Rev. 35, 151–163 (2015)Google Scholar
- 11.T. Zhang, L.B. Nanney, C. Luongo, L. Lamps, K.J. Heppner, R.N. DuBois, R.D. Beauchamp, Concurrent overexpression of cyclin D1 and cyclin-dependent kinase 4 (Cdk4) in intestinal adenomas from multiple intestinal neoplasia (Min) mice and human familial adenomatous polyposis patients. Cancer Res. 57, 169–175 (1997)PubMedGoogle Scholar
- 15.A. DeMichele, A.S. Clark, K.S. Tan, D.F. Heitjan, K. Gramlich, M. Gallagher, P. Lal, M. Feldman, P. Zhang, C. Colameco, D. Lewis, M. Langer, N. Goodman, S. Domchek, K. Gogineni, M. Rosen, K. Fox, P. O'Dwyer, CDK 4/6 inhibitor palbociclib (PD0332991) in Rb + advanced breast cancer: phase II activity, safety, and predictive biomarker assessment. Clin. Cancer Res. 21, 995–1001 (2015)CrossRefPubMedGoogle Scholar
- 16.M.A. Dickson, W.D. Tap, M.L. Keohan, S.P. D'Angelo, M.M. Gounder, C.R. Antonescu, J. Landa, L.X. Qin, D.D. Rathbone, M.M. Condy, Y. Ustoyev, A.M. Crago, S. Singer, G.K. Schwartz, Phase II trial of the CDK4 inhibitor PD0332991 in patients with advanced CDK4-amplified well-differentiated or dedifferentiated liposarcoma. J. Clin. Oncol. 31, 2024–2028 (2013)CrossRefPubMedPubMedCentralGoogle Scholar
- 17.F. Morschhauser, K. Bouabdallah, S. Stilgenbauer, C. Thieblemont, M. Wolf, S. de Guibert, F. Zettl, M. Hahka-Kemppinen, D.X. Wang, P. Brueck, Clinical activity of abemaciclib (LY2835219), a cell cycle inhibitor selective for CDK4 and CDK6, in patients with relapsed or refractory mantle cell lymphoma. Blood 124, 3067–3067 (2014)Google Scholar
- 18.J.R. Infante, G. Shapiro, P. Witteveen, J.F. Gerecitano, V. Ribrag, R. Chugh, I. Issa, A. Chakraborty, A. Matano and X. Zhao, In ASCO Annual Meeting Proceedings, p. 2528 (2014)Google Scholar
- 19.R.S. Finn, J.P. Crown, I. Lang, K. Boer, I.M. Bondarenko, S.O. Kulyk, J. Ettl, R. Patel, T. Pinter, M. Schmidt, Y. Shparyk, A.R. Thummala, N.L. Voytko, C. Fowst, X. Huang, S.T. Kim, S. Randolph, D.J. Slamon, The cyclin-dependent kinase 4/6 inhibitor palbociclib in combination with letrozole versus letrozole alone as first-line treatment of oestrogen receptor-positive, HER2-negative, advanced breast cancer (PALOMA-1/TRIO-18): a randomised phase 2 study. Lancet Oncol. 16, 25–35 (2015)CrossRefPubMedGoogle Scholar
- 20.D.W. Fry, P.J. Harvey, P.R. Keller, W.L. Elliott, M. Meade, E. Trachet, M. Albassam, X. Zheng, W.R. Leopold, N.K. Pryer, P.L. Toogood, Specific inhibition of cyclin-dependent kinase 4/6 by PD 0332991 and associated antitumor activity in human tumor xenografts. Mol. Cancer Ther. 3, 1427–1438 (2004)PubMedGoogle Scholar
- 21.J. Halder, C.N. Landen Jr., S.K. Lutgendorf, Y. Li, N.B. Jennings, D. Fan, G.M. Nelkin, R. Schmandt, M.D. Schaller, A.K. Sood, Focal adhesion kinase silencing augments docetaxel-mediated apoptosis in ovarian cancer cells. Clin. Cancer Res. 11, 8829–8836 (2005)CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Z. Duan, A.J. Feller, R.T. Penson, B.A. Chabner, M.V. Seiden, Discovery of differentially expressed genes associated with paclitaxel resistance using cDNA array technology: analysis of interleukin (IL) 6, IL-8, and monocyte chemotactic protein 1 in the paclitaxel-resistant phenotype. Clin. Cancer Res. 5, 3445–3453 (1999)PubMedGoogle Scholar
- 29.T.U. Ha, D.L. Segev, D. Barbie, P.T. Masiakos, T.T. Tran, D. Dombkowski, M. Glander, T.R. Clarke, H.K. Lorenzo, P.K. Donahoe, S. Maheswaran, Mullerian inhibiting substance inhibits ovarian cell growth through an Rb-independent mechanism. J. Biol. Chem. 275, 37101–37109 (2000)CrossRefPubMedGoogle Scholar
- 31.J.P. Leonard, A.S. LaCasce, M.R. Smith, A. Noy, L.R. Chirieac, S.J. Rodig, J.Q. Yu, S. Vallabhajosula, H. Schoder, P. English, D.S. Neuberg, P. Martin, M.M. Millenson, S.A. Ely, R. Courtney, N. Shaik, K.D. Wilner, S. Randolph, A.D. Van den Abbeele, S.Y. Chen-Kiang, J.T. Yap, G.I. Shapiro, Selective CDK4/6 inhibition with tumor responses by PD0332991 in patients with mantle cell lymphoma. Blood 119, 4597–4607 (2012)CrossRefPubMedGoogle Scholar
- 32.C.K. Ingemarsdotter, L.A. Tookman, A. Browne, K. Pirlo, R. Cutts, C. Chelela, K.F. Khurrum, E.Y. Leung, S. Dowson, L. Webber, I. Khan, D. Ennis, N. Syed, T.R. Crook, J.D. Brenton, M. Lockley, I.A. McNeish, Paclitaxel resistance increases oncolytic adenovirus efficacy via upregulated CAR expression and dysfunctional cell cycle control. Mol. Oncol. 9, 791–805 (2015)CrossRefPubMedGoogle Scholar
- 33.A.S. Clark, P.J. O'Dwyer, D. Heitjan, P. Lal, M.D. Feldman, M. Gallagher, C. Redlinger, C. Colameco, D. Lewis, K. Zafman, M. Langer, M.A. Rosen, K. Gogineni, A.R. Bradbury, S.M. Domchek, K.R. Fox, A. DeMichele, A phase I trial of palbociclib and paclitaxel in metastatic breast cancer. J. Clin. Oncol. 32, 5 (2014)Google Scholar
- 40.S.N. Kolomeichuk, D.T. Terrano, C.S. Lyle, K. Sabapathy, T.C. Chambers, Distinct signaling pathways of microtubule inhibitors--vinblastine and Taxol induce JNK-dependent cell death but through AP-1-dependent and AP-1-independent mechanisms, respectively. FEBS J. 275, 1889–1899 (2008)CrossRefPubMedGoogle Scholar
- 43.D.W. Fry, D.C. Bedford, P.H. Harvey, A. Fritsch, P.R. Keller, Z. Wu, E. Dobrusin, W.R. Leopold, A. Fattaey, M.D. Garrett, Cell cycle and biochemical effects of PD 0183812. A potent inhibitor of the cyclin D-dependent kinases CDK4 and CDK6. J. Biol. Chem. 276, 16617–16623 (2001)CrossRefPubMedGoogle Scholar
- 44.B. Taylor-Harding, P.J. Aspuria, H. Agadjanian, D.J. Cheon, T. Mizuno, D. Greenberg, J.R. Allen, L. Spurka, V. Funari, E. Spiteri, Q. Wang, S. Orsulic, C. Walsh, B.Y. Karlan, W.R. Wiedemeyer, Cyclin E1 and RTK/RAS signaling drive CDK inhibitor resistance via activation of E2F and ETS. Oncotarget 6, 696–714 (2015)CrossRefPubMedGoogle Scholar
- 45.G.E. Konecny, B. Winterhoff, T. Kolarova, J. Qi, K. Manivong, J. Dering, G. Yang, M. Chalukya, H.J. Wang, L. Anderson, K.R. Kalli, R.S. Finn, C. Ginther, S. Jones, V.E. Velculescu, D. Riehle, W.A. Cliby, S. Randolph, M. Koehler, L.C. Hartmann, D.J. Slamon, Expression of p16 and retinoblastoma determines response to CDK4/6 inhibition in ovarian cancer. Clin. Cancer Res. 17, 1591–1602 (2011)CrossRefPubMedPubMedCentralGoogle Scholar