12-Deoxyphorbol Esters Induce Growth Arrest and Apoptosis in Human Lung Cancer A549 Cells Via Activation of PKC-δ/PKD/ERK Signaling Pathway
Abstract
:1. Introduction
2. Results
2.1. GRC-2 Is More Potent than Prostratin in Inhibiting the Cell Viability of A549 Cancer Cells by Inducing Cell Cycle Arrest and Apoptosis
2.2. 12-Deoxyphorbol Esters Cause Activation of PKCs in A549 Cells
2.3. Knockdown of PKC-δ/PKD Reverses 12-Deoxyphorbol Esters-Induced Cytotoxicity against A549 Cancer Cells
2.4. The ERK Signaling Pathway is the Major Mediator of PKC-δ/D-Dependent Cytotoxicity of the 12-Deoxyphorbol Esters
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Lines
4.3. Cell Viability Assay
4.4. Transfection
4.5. Western Blotting
4.6. Immunofluorescence Staining
4.7. Apoptosis Assay
4.8. Cell Cycles Assay
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
PKC | Protein kinase C |
NSCLC | Human non-small cell lung cancer |
PKD | Protein kinase D |
ERK | Extracellular signal-related kinase |
TPA | Phorbol 12-myristate 13-acetate |
DAG | Diacylglycerol |
MTT | 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
PI | Propidium iodide |
DAPI | 4′,6-diamidino-2-phenylindole |
DMSO | Dimethyl sulfoxide |
MAP | Mitogen-activated protein |
CDK1 | Cyclin-dependent kinase-1 |
HIV | Human immunodeficiency virus |
References
- Vasas, A.; Hohmann, J. Euphorbia diterpenes: Isolation, structure, biological activity, and synthesis (2008–2012). Chem. Rev. 2014, 114, 8579–8612. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wang, H.B.; Wang, X.Y.; Liu, L.P.; Qin, G.W.; Kang, T.G. Tigliane diterpenoids from the Euphorbiaceae and Thymelaeaceae families. Chem. Rev. 2015, 115, 2975–3011. [Google Scholar] [CrossRef] [PubMed]
- Szallasi, Z.; Krsmanovic, L.; Blumberg, P.M. Nonpromoting 12-deoxyphorbol 13-esters inhibit phorbol 12-myristate 13-acetate induced tumor promotion in CD-1 mouse skin. Cancer Res. 1993, 53, 2507–2512. [Google Scholar] [PubMed]
- Zayed, S.; Sorg, B.; Hecker, E. Structure activity relations of polyfunctional diterpenes of the tigliane type, VI. Irritant and tumor promoting activities of semisynthetic mono and diesters of 12-deoxyphorbol. Planta Med. 1984, 50, 65–69. [Google Scholar] [CrossRef] [PubMed]
- Wang, Y.; Tang, C.; Yao, S.; Lai, H.; Li, R.; Xu, J.; Wang, Q.; Fan, X.X.; Wu, Q.B.; Leung, E.L.; et al. Discovery of a novel protein kinase C activator from Croton tiglium for inhibition of non-small cell lung cancer. Phytomedicine 2019, 65, 153100. [Google Scholar] [CrossRef] [PubMed]
- Alotaibi, D.; Amara, S.; Johnson, T.L.; Tiriveedhi, V. Potential anticancer effect of prostratin through SIK3 inhibition. Oncol. Lett. 2018, 15, 3252–3258. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Shen, X.; Xiong, G.L.; Jing, Y.; Xiao, H.; Cui, Y.; Zhang, Y.F.; Shan, Y.J.; Xing, S.; Yang, M.; Liu, X.L.; et al. The protein kinase C agonist prostratin induces differentiation of human myeloid leukemia cells and enhances cellular differentiation by chemotherapeutic agents. Cancer Lett. 2015, 356, 686–696. [Google Scholar] [CrossRef]
- Kostovic, K.; Gulin, S.J.; Mokos, Z.B.; Ceovic, R. Topical Ingenol Mebutate: A New Treatment Modality for Multiple Actinic Keratoses and Field Cancerization. Anti-Cancer Agents Med. Chem. 2017, 17, 1304–1311. [Google Scholar] [CrossRef]
- Isakov, N. Protein kinase C (PKC) isoforms in cancer, tumor promotion and tumor suppression. Semin. Cancer Biol. 2018, 48, 36–52. [Google Scholar] [CrossRef]
- Dempsey, E.C.; Newton, A.C.; Mochly-Rosen, D.; Fields, A.P.; Reyland, M.E.; Insel, P.A.; Messing, R.O. Protein kinase C isozymes and the regulation of diverse cell responses. Am. J. Physiol. Lung Cell Mol. Physiol. 2000, 279, L429–L438. [Google Scholar] [CrossRef]
- Koivunen, J.; Aaltonen, V.; Peltonen, J. Protein kinase C (PKC) family in cancer progression. Cancer Lett. 2006, 235, 1–10. [Google Scholar] [CrossRef] [PubMed]
- Zhao, M.; Xia, L.; Chen, G.Q. Protein kinase cdelta in apoptosis: A brief overview. Arch. Immunol. Ther. Exp. 2012, 60, 361–372. [Google Scholar] [CrossRef] [PubMed]
- Newton, A.C. Protein kinase C as a tumor suppressor. Semin. Cancer Biol. 2018, 48, 18–26. [Google Scholar] [CrossRef]
- Newton, A.C.; Brognard, J. Reversing the Paradigm: Protein Kinase C as a Tumor Suppressor. Trends Pharmacol. Sci. 2017, 38, 438–447. [Google Scholar] [CrossRef] [Green Version]
- Fu, Y.; Rubin, C.S. Protein kinase D: Coupling extracellular stimuli to the regulation of cell physiology. EMBO Rep. 2011, 12, 785–796. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rozengurt, E.; Rey, O.; Waldron, R.T. Protein kinase D signaling. J. Biol. Chem. 2005, 280, 13205–13208. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Li, J.; O’Connor, K.L.; Hellmich, M.R.; Greeley, G.H., Jr.; Townsend, C.M., Jr.; Evers, B.M. The role of protein kinase D in neurotensin secretion mediated by protein kinase C-alpha/-delta and Rho/Rho kinase. J. Biol. Chem. 2004, 279, 28466–28474. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wong, C.; Jin, Z.G. Protein kinase C-dependent protein kinase D activation modulates ERK signal pathway and endothelial cell proliferation by vascular endothelial growth factor. J. Biol. Chem. 2005, 280, 33262–33269. [Google Scholar] [CrossRef] [Green Version]
- Roy, A.; Ye, J.; Deng, F.; Wang, Q.J. Protein kinase D signaling in cancer: A friend or foe? Biochim. Biophys. Acta. Rev. Cancer 2017, 1868, 283–294. [Google Scholar] [CrossRef]
- Tsai, J.-Y.; Rédei, D.R.; Forgo, P.; Li, Y.; Vasas, A.; Hohmann, J.; Wu, C.-C. Isolation of Phorbol Esters from Euphorbia grandicornis and Evaluation of Protein Kinase C-and Human Platelet-Activating Effects of Euphorbiaceae Diterpenes. J. Nat. Prod. 2016, 79, 2658–2666. [Google Scholar] [CrossRef]
- Wang, Q.J. PKD at the crossroads of DAG and PKC signaling. Trends Pharmacol. Sci. 2006, 27, 317–323. [Google Scholar] [CrossRef]
- Matsumoto, E.; Hatanaka, M.; Bohgaki, M.; Maeda, S. PKC pathway and ERK/MAPK pathway are required for induction of cyclin D1 and p21Waf1 during 12-o-tetradecanoylphorbol 13-acetate-induced differentiation of myeloleukemia cells. Kobe J. Med. Sci. 2006, 52, 181. [Google Scholar]
- Park, K.S.; Jeon, S.H.; Oh, J.W.; Choi, K.Y. p21Cip/WAF1 activation is an important factor for the ERK pathway dependent anti-proliferation of colorectal cancer cells. Exp. Mol. Med. 2004, 36, 557–562. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tanaka, Y.; Gavrielides, M.V.; Mitsuuchi, Y.; Fujii, T.; Kazanietz, M.G. Protein kinase C promotes apoptosis in LNCaP prostate cancer cells through activation of p38 MAPK and inhibition of the Akt survival pathway. J. Biol. Chem. 2003, 278, 33753–33762. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Todd, D.E.; Densham, R.M.; Molton, S.A.; Balmanno, K.; Newson, C.; Weston, C.R.; Garner, A.P.; Scott, L.; Cook, S.J. ERK1/2 and p38 cooperate to induce a p21CIP1-dependent G1 cell cycle arrest. Oncogene 2004, 23, 3284–3295. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kreis, N.N.; Louwen, F.; Yuan, J. Less understood issues: p21(Cip1) in mitosis and its therapeutic potential. Oncogene 2015, 34, 1758–1767. [Google Scholar] [CrossRef]
- Andersen, R.J.; Ntie-Kang, F.; Tietjen, I. Natural product-derived compounds in HIV suppression, remission, and eradication strategies. Antiviral Res. 2018, 158, 63–77. [Google Scholar] [CrossRef]
- Miana, G.A.; Riaz, M.; Shahzad-ul-Hussan, S.; Paracha, R.Z.; Paracha, U.Z. Prostratin: An Overview. Mini Rev. Med. Chem. 2015, 15, 1122–1130. [Google Scholar] [CrossRef]
- Adwan, T.S.; Ohm, A.M.; Jones, D.N.; Humphries, M.J.; Reyland, M.E. Regulated binding of importin-alpha to protein kinase Cdelta in response to apoptotic signals facilitates nuclear import. J. Biol. Chem. 2011, 286, 35716–35724. [Google Scholar] [CrossRef] [Green Version]
- DeVries-Seimon, T.A.; Ohm, A.M.; Humphries, M.J.; Reyland, M.E. Induction of apoptosis is driven by nuclear retention of protein kinase C delta. J. Biol. Chem. 2007, 282, 22307–22314. [Google Scholar] [CrossRef] [Green Version]
- Unni, A.M.; Harbourne, B.; Oh, M.H.; Wild, S.; Ferrarone, J.R.; Lockwood, W.W.; Varmus, H. Hyperactivation of ERK by multiple mechanisms is toxic to RTK-RAS mutation-driven lung adenocarcinoma cells. ELife 2018, 7, e33718. [Google Scholar] [CrossRef] [PubMed]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Tsai, J.-Y.; Rédei, D.; Hohmann, J.; Wu, C.-C. 12-Deoxyphorbol Esters Induce Growth Arrest and Apoptosis in Human Lung Cancer A549 Cells Via Activation of PKC-δ/PKD/ERK Signaling Pathway. Int. J. Mol. Sci. 2020, 21, 7579. https://doi.org/10.3390/ijms21207579
Tsai J-Y, Rédei D, Hohmann J, Wu C-C. 12-Deoxyphorbol Esters Induce Growth Arrest and Apoptosis in Human Lung Cancer A549 Cells Via Activation of PKC-δ/PKD/ERK Signaling Pathway. International Journal of Molecular Sciences. 2020; 21(20):7579. https://doi.org/10.3390/ijms21207579
Chicago/Turabian StyleTsai, Ju-Ying, Dóra Rédei, Judit Hohmann, and Chin-Chung Wu. 2020. "12-Deoxyphorbol Esters Induce Growth Arrest and Apoptosis in Human Lung Cancer A549 Cells Via Activation of PKC-δ/PKD/ERK Signaling Pathway" International Journal of Molecular Sciences 21, no. 20: 7579. https://doi.org/10.3390/ijms21207579