Proline-Dependent Induction of Apoptosis in Oral Squamous Cell Carcinoma (OSCC)—The Effect of Celecoxib
Abstract
:1. Introduction
2. Results
2.1. Proline Concentration in Oral Cancer Tissue in Comparison to Normal Oral Mucosa
2.2. Expression of Selected Proteins (PRODH/POX, PPAR-γ, HIF-1α) Involved in Proline Metabolism and Important in the Process of Apoptosis—Tissue Material
2.3. Differences in the Expression of PRODH/POX, PPARγ and HIF-1α according to the Tumor Malignancy
2.4. Viability Assay and Proliferation Assay Cell Culture
2.5. Expression of PRODH/POX, PPARγ, and HIF-1α in CAL-27 Cells Incubated with Various Concentrations of Celecoxib
2.6. Evaluation of Apoptosis—Expression of Caspases-3 and 9 in CAL-27 Cancer Cells after Incubation with Celecoxib
3. Discussion
- -
- -
- -
- HIF-1α (hypoxia-inducible factor 1-alpha) is a transcription factor specifically activated by low oxygen concentration in the cellular environment. It is an indispensable component of the transcriptional response of tumours to hypoxia. It also influences an increase in the expression of key factors determining cancer development, e.g., VEGF, COX-2, NF-κB [10,29,30].
4. Materials and Methods
4.1. Patients and Tissues
4.2. Evaluation of Proline Concentration in Tissue Material—Liquid Chromatography Combined with Tandem Mass Spectrometry (HPLC-MS/MS)
4.3. Expression of Selected Proteins by Western Immunoblot—Tissues
4.4. Cell Culture
4.5. Cell Viability Assay—MTT Test
4.6. Proliferation Assay—DNA Biosynthesis
4.7. Expression of Selected Proteins by Western Immunoblot—Cell Lines
4.8. Evaluation of Apoptosis—Immunofluorescence Microscopy
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Rivera, C. Essentials of oral cancer. Int. J. Clin. Exp. Pathol. 2015, 8, 11884–11894. [Google Scholar] [PubMed]
- Bray, F.; Ferlay, J.; Soerjomataram, I.; Siegel, R.L.; Torre, L.A.; Jemal, A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 2018, 68, 394–424. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gupta, B.; Johnson, N.W.; Kumar, N. Global epidemiology of head and neck cancers: A continuing challenge. Oncology 2016, 91, 13–23. [Google Scholar] [CrossRef] [PubMed]
- Gupta, N.; Gupta, R.; Acharya, A.K.; Patthi, B.; Goud, V.; Reddy, S.; Garg, A.; Singla, A. Changing trends in oral cancer—A global scenario. Nepal J. Epidemiol. 2016, 6, 613–619. [Google Scholar] [CrossRef]
- Shield, K.D.; Ferlay, J.; Jemal, A.; Sankaranarayanan, R.; Chaturvedi, A.K.; Bray, F.; Soerjomataram, I. The global incidence of lip, oral cavity, and pharyngeal cancers by subsite in 2012. Cancer J. Clin. 2017, 67, 51–64. [Google Scholar] [CrossRef]
- Colevas, A.D.; Yom, S.S.; Pfister, D.G.; Spencer, S.; Adelstein, D.; Adkins, D.; Brizel, D.M.; Burtness, B.; Busse, P.M.; Caudell, J.J.; et al. NCCN Guidelines Insights: Head and Neck Cancers, Version 1.2018. J. Natl Compr. Canc. Netw. 2018, 16, 479–490. [Google Scholar] [CrossRef] [Green Version]
- Huang, S.H.; O’Sullivan, B. Oral cancer: Current role of radiotherapy and chemotherapy. Med. Oral Patol. Oral Cir. Bucal 2013, 18, e233–e240. [Google Scholar] [CrossRef]
- Vigneswaran, N.; Williams, M.D. Epidemiological trends in head and neck cancer and aids in diagnosis. Oral Maxillofac. Surg. Clin. N. Am. 2014, 26, 123–141. [Google Scholar] [CrossRef]
- Morgan, A.A.; Rubenstein, E. Proline: The distribution, frequency, positioning, and common functional roles of proline and polyproline sequences in the human proteome. PLoS ONE 2013, 8, e53785. [Google Scholar] [CrossRef] [Green Version]
- Phang, J.M.; Liu, W.; Zabirnyk, O. Proline metabolism and microenvironmental stress. Annu. Rev. Nutr. 2010, 30, 441–463. [Google Scholar] [CrossRef] [Green Version]
- Togashi, Y.; Arao, T.; Kato, H.; Matsumoto, K.; Terashima, M.; Hayashi, H.; de Velasco, M.A.; Fujita, Y.; Kimura, H.; Yasuda, T.; et al. Frequent amplification of ORAOV1 gene in esophageal squamous cell cancer promotes an aggressive phenotype via proline metabolism and ROS production. Oncotarget 2014, 5, 2962–2973. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Phang, J.M.; Liu, W.; Hancock, C.N.; Fischer, J.W. Proline metabolism and cancer: Emerging links to glutamine and collagen. Curr. Opin. Clin. Nutr. Metab. Care 2015, 18, 71–77. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Phang, J.M.; Liu, W. Proline metabolism and cancer. Front. Biosci. 2012, 17, 1835–1845. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Maxwell, S.A.; Rivera, A. Proline oxidase induces apoptosis in tumor cells, and its expression is frequently absent or reduced in renal carcinomas. J. Biol. Chem. 2003, 278, 9784–9789. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liu, Y.; Borchert, G.L.; Donald, S.P.; Diwan, B.A.; Anver, M.; Phang, M. Proline oxidase functions as a mitochondrial tumor suppressor in human cancers. Cancer Res. 2009, 69, 6414–6422. [Google Scholar] [CrossRef] [Green Version]
- Coussens, L.M.; Werb, Z. Inflammation and cancer. Nature 2002, 420, 860–867. [Google Scholar] [CrossRef]
- Harris, R.E.; Beebe, J.; Alshafie, G.A. Reduction in cancer risk by selective and nonselective cyclooxygenase-2 (COX-2) inhibitors. J. Exp. Pharmacol. 2012, 4, 91–96. [Google Scholar] [CrossRef] [Green Version]
- Fujii, R.; Imanishi, Y.; Shibata, K.; Sakai, N.; Sakamoto, K.; Shigetomi, S.; Habu, N.; Otsuka, K.; Sato, Y.; Watanabe, Y.; et al. Restoration of E-cadherin expression by selective Cox-2 inhibition and the clinical relevance of the epithelial-to-mesenchymal transition in head and neck squamous cell carcinoma. J. Exp. Clin. Cancer Res. 2014, 33, 40. [Google Scholar] [CrossRef] [Green Version]
- Sacchetti, A. Cancer cell killing by Celecoxib: Reality or just in vitro precipitation-related artifact? J. Cell. Biochem. 2013, 114, 1434–1444. [Google Scholar] [CrossRef]
- Komatsu, Y.; Hibi, K.; Kodera, Y.; Akiyama, S.; Ito, K.; Nakao, A. TAOS1, a novel marker for advanced esophageal squamous cell carcinoma. Anticancer Res. 2006, 26, 2029–2032. [Google Scholar]
- Krishnan, N.; Dickman, M.B.; Becker, D.F. Proline modulates the intracellular redox environment and protects mammalian cells against oxidative stress. Free Radic. Biol. Med. 2008, 44, 671–681. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Natarajan, S.K.; Zhu, W.; Liang, X.; Zhang, L.; Demers, A.J.; Zimmerman, M.C.; Simpson, M.A.; Becker, D.F. Proline dehydrogenase is essential for proline protection against hydrogen peroxide-induced cell death. Free Radic. Biol. Med. 2012, 53, 1181–1191. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liu, W.; Le, A.; Hancock, C.; Lane, A.N.; Dang, C.V.; Fan, T.W.; Phang, J.M. Reprogramming of proline and glutamine metabolism contributes to the proliferative and metabolic responses regulated by oncogenic transcription factor c-MYC. Proc. Natl. Acad. Sci. USA 2012, 109, 8983–8988. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liu, W.; Hancock, C.N.; Fischer, J.W.; Harman, M.; Phang, J.M. Proline biosynthesis augments tumor cell growth and aerobic glycolysis: Involvement of pyridine nucleotides. Sci. Rep. 2015, 5, 17206. [Google Scholar] [CrossRef] [Green Version]
- Vermeersch, K.A.; Wang, L.; Mezencev, R.; McDonald, J.F.; Styczynski, M.P. OVCAR-3 spheroid-derived cells display distinct metabolic profiles. PLoS ONE 2015, 10, e0118262. [Google Scholar] [CrossRef] [Green Version]
- Kononczuk, J.; Czyzewska, U.; Moczydlowska, J.; Surażyński, A.; Palka, J.; Miltyk, W. Proline oxidase (POX) as a target for cancer therapy. Curr. Drug Targets 2015, 16, 1464–1469. [Google Scholar] [CrossRef]
- Phang, J.M.; Pandhare, J.; Zabirnyk, O.; Liu, Y. PPAR and proline oxidase in cancer. PPAR Res. 2008, 2008, 542694. [Google Scholar] [CrossRef] [Green Version]
- Bonofiglio, D.; Aquila, S.; Catalano, S.; Gabriele, S.; Belmonte, M.; Middea, E.; Qi, H.; Morelli, C.; Gentile, M.; Maggiolini, M.; et al. Peroxisome proliferator-activated receptor-gamma activates p53 gene promoter binding to the nuclear factor-kappaB sequence in human MCF7 breast cancer cells. Mol. Endocrinol. 2006, 20, 3083–3092. [Google Scholar] [CrossRef]
- Pugh, C.W.; Ratcliffe, P. Regulation of angiogenesis by hypoxia: Role of the HIF system. Nat. Med. 2003, 9, 677–684. [Google Scholar] [CrossRef]
- Quintero, M.; Mackenzie, N.; Brennan, P.A. Hypoxia-inducible factor 1 (HIF-1) in cancer. Eur. J. Surg. Oncol. 2004, 30, 465–468. [Google Scholar] [CrossRef]
- Jendrossek, V. Targeting apoptosis pathways by Celecoxib in cancer. Cancer Lett. 2013, 332, 313–324. [Google Scholar] [CrossRef]
- Chiang, S.L.; Velmurugan, B.K.; Chung, C.M.; Lin, S.H.; Wang, Z.H.; Hua, C.H.; Tsai, M.H.; Kuo, T.M.; Yeh, K.T.; Chang, P.Y.; et al. Preventive effect of celecoxib use against cancer progression and occurrence of oral squamous cell carcinoma. Sci. Rep. 2017, 7, 6235. [Google Scholar] [CrossRef]
- Xiang, H.G.; Xie, X.; Hu, F.Q.; Xiao, H.B.; Zhang, W.J.; Chen, L. Cyclooxygenase-2 inhibition as a strategy for treating gastric adenocarcinoma. Oncol. Rep. 2014, 32, 1140–1148. [Google Scholar] [CrossRef] [PubMed]
- Liu, Y.; Borchert, G.L.; Surazynski, A.; Phang, J.M. Proline oxidase, a p53-induced gene, targets COX-2/PGE2 signaling to induce apoptosis and inhibit tumor growth in colorectal cancers. Oncogene 2008, 27, 6729–6737. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Soo, R.A.; Wu, J.; Aggarwal, A.; Tao, Q.; Hsieh, W.; Putti, T.; Tan, K.B.; Low, J.S.; Lai, Y.F.; Mow, B.; et al. Celecoxib reduces microvessel density in patients treated with nasopharyngeal carcinoma and induces changes in gene expression. Ann. Oncol. 2006, 17, 1625–1630. [Google Scholar] [CrossRef] [PubMed]
- Masferrer, J.L.; Leahy, K.M.; Koki, A.T.; Zweifel, B.S.; Settle, S.L.; Woerner, B.M.; Edwards, D.A.; Flickinger, A.G.; Moore, R.J.; Seibert, K. Antiangiogenic and antitumor activities of cyclooxygenase-2 inhibitors. Cancer Res. 2000, 60, 1306–1311. [Google Scholar]
- Williams, C.S.; Watson, A.J.M.; Sheng, H.; Helou, R.; Shao, J.; Dubois, R.N. Celecoxib prevents tumor growth in vivo without toxicity to normal gut: Lack of correlation between in vitro and in vivo models. Cancer Res. 2000, 60, 6045–6051. [Google Scholar]
- Leahy, K.M.; Ornberg, R.L.; Wang, Y.; Zweifel, B.S.; Koki, A.T.; Masferrer, J.L. Cyclooxygenase-2 inhibition by celecoxib reduces proliferation and induces apoptosis in angiogenic endothelial cell in vivo. Cancer Res. 2002, 62, 625–631. [Google Scholar]
- Toloczko-Iwaniuk, N.; Dziemianczyk-Pakiela, D.; Nowaszewska, B.K.; Celinska-Janowicz, K.; Miltyk, W. Celecoxib in cancer therapy and prevention—Review. Curr. Drug Targets 2019, 20, 302–315. [Google Scholar] [CrossRef]
- Phang, J.M.; Liu, W.; Hancock, C.; Christian, K.J. The proline regulatory axis and cancer. Front. Oncol. 2012, 2, 60. [Google Scholar] [CrossRef] [Green Version]
- Phang, J.M.; Donald, S.P.; Pandhare, J.; Liu, Y. The metabolism of proline, a stress substrate, modulates carcinogenic pathways. Amino Acids 2008, 35, 681–690. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Brentnall, M.; Rodriguez-Menocal, M.; Ladron De Guevara, R.; Cepero, E.; Boise, L.H. Caspase-9, caspase-3 and caspase-7 have distinct roles duringN intrinsic apoptosis. BMC Cell Biol. 2013, 14, 32. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Allan, L.A.; Clarke, P.R. Apoptosis and autophagy: Regulation of caspase-9 by phosphorylation. FEBS J. 2009, 276, 6063–6073. [Google Scholar] [CrossRef] [Green Version]
- Elmore, S. Apoptosis: A review of programmed cell death. Toxicol. Pathol. 2007, 35, 495–516. [Google Scholar] [CrossRef] [PubMed]
- Porter, A.G.; Jänicke, R.U. Emerging roles of caspase-3 in apoptosis. Cell Death Differ. 1999, 6, 99–104. [Google Scholar] [CrossRef]
- Union for International Cancer Control (UICC). TNM Classification of Malignant Tumours, 8th ed.; Brierley, J.D., Gospodarowicz, M.K., Wittekind, C., Eds.; Wiley–Blackwell: Hoboken, NJ, USA, 2016. [Google Scholar]
- Akhter, M.; Hossain, S.; Rahman, Q.B.; Molla, M.R. A study on histological grading of oral squamous cell carcinoma and its co-relationship with regional metastasis. J. Oral Maxillofac. Pathol. 2011, 15, 168–176. [Google Scholar] [CrossRef] [Green Version]
- Matysiak, J.; Dereziński, P.; Klupczynska, A.; Matysiak, J.; Kaczmarek, E.; Kokot, Z.J. Effects of a honeybee sting on the serum free amino acid profile in humans. PLoS ONE 2014, 9, e103533. [Google Scholar] [CrossRef] [Green Version]
- Plewa, S.; Horała, A.; Dereziński, P.; Klupczynska, A.; Nowak-Markwitz, E.; Matysiak, J.; Kokot, Z.J. Usefulness of amino acid profiling in ovarian cancer screening with special emphasis on their role in cancerogenesis. Int. J. Mol. Sci. 2017, 18, 2727. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lowry, O.H.; Rosebrough, N.J.; Farr, A.L.; Randall, R.J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 1951, 193, 265–275. [Google Scholar]
- Laemmli, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970, 227, 680–685. [Google Scholar] [CrossRef]
- Carmichael, J.; DeGraff, W.G.; Gazdar, A.F.; Minna, J.D.; Mitchell, J.B. Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of chemosensitivity testing. Cancer Res. 1987, 47, 936–942. [Google Scholar] [PubMed]
© 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
Tołoczko-Iwaniuk, N.; Dziemiańczyk-Pakieła, D.; Celińska-Janowicz, K.; Zaręba, I.; Klupczyńska, A.; Kokot, Z.J.; Nowaszewska, B.K.; Reszeć, J.; Borys, J.; Miltyk, W. Proline-Dependent Induction of Apoptosis in Oral Squamous Cell Carcinoma (OSCC)—The Effect of Celecoxib. Cancers 2020, 12, 136. https://doi.org/10.3390/cancers12010136
Tołoczko-Iwaniuk N, Dziemiańczyk-Pakieła D, Celińska-Janowicz K, Zaręba I, Klupczyńska A, Kokot ZJ, Nowaszewska BK, Reszeć J, Borys J, Miltyk W. Proline-Dependent Induction of Apoptosis in Oral Squamous Cell Carcinoma (OSCC)—The Effect of Celecoxib. Cancers. 2020; 12(1):136. https://doi.org/10.3390/cancers12010136
Chicago/Turabian StyleTołoczko-Iwaniuk, Natalia, Dorota Dziemiańczyk-Pakieła, Katarzyna Celińska-Janowicz, Ilona Zaręba, Agnieszka Klupczyńska, Zenon J. Kokot, Beata Klaudia Nowaszewska, Joanna Reszeć, Jan Borys, and Wojciech Miltyk. 2020. "Proline-Dependent Induction of Apoptosis in Oral Squamous Cell Carcinoma (OSCC)—The Effect of Celecoxib" Cancers 12, no. 1: 136. https://doi.org/10.3390/cancers12010136