Increased killing of SCCVII squamous cell carcinoma cells after the combination of Pc 4 photodynamic therapy and dasatinib is associated with enhanced caspase-3 activity and ceramide synthase 1 upregulation

Separovic,Duska; Breen,Paul; Boppana,Nithin   B.; Van Buren,Eric; Joseph,Nicholas; Kraveka,Jacqueline   M.; Rahmaniyan,Mehrdad; Li,Li; Gudz,Tatyana  I.; Bielawska,Alicja; Bai,Aiping; Bielawski,Jacek; Pierce,Jason   S.; Korbelik,Mladen

doi:10.3892/ijo.2013.2132

Increased killing of SCCVII squamous cell carcinoma cells after the combination of Pc 4 photodynamic therapy and dasatinib is associated with enhanced caspase-3 activity and ceramide synthase 1 upregulation

Authors:
- Duska Separovic
- Paul Breen
- Nithin B. Boppana
- Eric Van Buren
- Nicholas Joseph
- Jacqueline M. Kraveka
- Mehrdad Rahmaniyan
- Li Li
- Tatyana I. Gudz
- Alicja Bielawska
- Aiping Bai
- Jacek Bielawski
- Jason S. Pierce
- Mladen Korbelik
View Affiliations

Affiliations: Department of Pharmaceutical Sciences, Eugene Applebaum College of Pharmacy and Health Sciences, Wayne State University, Detroit, MI 48201, USA, Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA, Department of Pediatrics, Division of Hematology-Oncology, Charles Darby Children's Research Institute, and Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425, USA, Ralph H. Johnson Veterans Affairs Medical Center, and Department of Neuroscience, Medical University of South Carolina, Charleston, SC 29425, USA, Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC 29425, USA, British Columbia Cancer Agency, Vancouver, BC V5Z 1L3, Canada
Published online on: October 9, 2013 https://doi.org/10.3892/ijo.2013.2132
Pages: 2064-2072

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Photodynamic therapy (PDT) is not always effective as an anticancer treatment, therefore, PDT is combined with other anticancer agents for improved efficacy. The combination of dasatinib and PDT with the silicone phthalocyanine photosensitizer Pc 4 was assessed for increased killing of SCCVII mouse squamous cell carcinoma cells, a preclinical model of head and neck squamous cell carcinoma, using apoptotic markers and colony formation as experimental end-points. Because each of these treatments regulates the metabolism of the sphingolipid ceramide, their effects on mRNA levels of ceramide synthase, a ceramide-producing enzyme, and the sphingolipid profile were determined. PDT + dasatinib induced an additive loss of clonogenicity. Unlike PDT alone or PDT + dasatinib, dasatinib induced zVAD-fmk-dependent cell killing. PDT or dasatinib-induced caspase-3 activation was potentiated after the combination. PDT alone induced mitochondrial depolarization, and the effect was inhibited after the combination. Annexin V+ and propidium iodide+ cells remained at control levels after treatments. In contrast to PDT alone, dasatinib induced upregulation of ceramide synthase 1 mRNA, and the effect was enhanced after the combination. Dasatinib induced a modest increase in C20:1- and C22-ceramide but had no effect on total ceramide levels. PDT increased the levels of 12 individual ceramides and total ceramides, and the addition of dasatinib did not affect these increases. PDT alone decreased substantially sphingosine levels and inhibited the activity of acid ceramidase, an enzyme that converts ceramide to sphingosine. The data suggest that PDT-induced increases in ceramide levels do not correlate with ceramide synthase mRNA levels but rather with inhibition of ceramidase. Cell killing was zVAD-fmk-sensitive after dasatinib but not after either PDT or the combination and enhanced cell killing after the combination correlated with potentiated caspase-3 activation and upregulation of ceramide synthase 1 mRNA but not the production of ceramide. The data imply potential significance of the combination for cancer treatment.

View Figures

View References

1.	Agostinis P, Berg K, Cengel KA, Foster TH, Girotti AW, Gollnick SO, Hahn SM, Hamblin MR, Juzeniene A, Kessel D, Korbelik M, Moan J, Mroz P, Nowis D, Piette J, Wilson BC and Golab J: Photodynamic therapy of cancer: an update. CA Cancer J Clin. 61:250–281. 2011. View Article : Google Scholar : PubMed/NCBI
2.	Wei G, Rafiyath S and Liu D: First-line treatment for chronic myeloid leukemia: dasatinib, nilotinib, or imatinib. J Hematol Oncol. 3:472010. View Article : Google Scholar : PubMed/NCBI
3.	Brooks HD, Glisson BS, Bekele BN, Johnson FM, Ginsberg LE, El-Naggar A, Culotta KS, Takebe N, Wright J, Tran HT and Papadimitrakopoulou VA: Phase 2 study of dasatinib in the treatment of head and neck squamous cell carcinoma. Cancer. 117:2112–2119. 2012. View Article : Google Scholar : PubMed/NCBI
4.	Kopetz S, Lesslie DP, Dallas NA, Park SI, Johnson M, Parikh NU, Kim MP, Abbruzzese JL, Ellis LM, Chandra J and Gallick GE: Synergistic activity of the SRC family kinase inhibitor dasatinib and oxaliplatin in colon carcinoma cells is mediated by oxidative stress. Cancer Res. 69:3842–3849. 2009. View Article : Google Scholar : PubMed/NCBI
5.	Pichot CS, Hartig SM, Xia L, Arvanitis C, Monisvais D, Lee FY, Frost JA and Corey SJ: Dasatinib synergizes with doxorubicin to block growth, migration, and invasion of breast cancer cells. Br J Cancer. 101:38–47. 2009. View Article : Google Scholar : PubMed/NCBI
6.	Haura EB, Tanvetyanon T, Chiappori A, Williams C, Simon G, Antonia S, Gray J, Litschauer S, Tetteh L, Neuger A, Song L, Rawal B, Schell MJ and Bepler G: Phase I/II study of the Src inhibitor dasatinib in combination with erlotinib in advanced non-small-cell lung cancer. J Clin Oncol. 28:1387–1394. 2010. View Article : Google Scholar : PubMed/NCBI
7.	Raju U, Riesterer O, Wang ZQ, Molkentine DP, Molkentine JM, Johnson FM, Glisson B, Milas L and Ang KK: Dasatinib, a multi-kinase inhibitor increased radiation sensitivity by interfering with nuclear localization of epidermal growth factor receptor and by blocking DNA repair pathways. Radiother Oncol. 105:241–249. 2012. View Article : Google Scholar
8.	Adan-Gokbulut A, Kartal-Yandim M, Iskender G and Baran Y: Novel agents targeting bioactive sphingolipids for the treatment of cancer. Curr Med Chem. 20:108–122. 2013. View Article : Google Scholar : PubMed/NCBI
9.	Hannun YA and Obeid LM: Many ceramides. J Biol Chem. 286:27855–27862. 2011. View Article : Google Scholar : PubMed/NCBI
10.	Saddoughi SA and Ogretmen B: Diverse functions of ceramide in cancer cell death and proliferation. Adv Cancer Res. 117:37–58. 2013. View Article : Google Scholar : PubMed/NCBI
11.	Separovic D, Breen P, Joseph N, Bielawski J, Pierce JS, Van Buren E and Gudz TI: Ceramide synthase 6 knockdown suppresses apoptosis after photodynamic therapy in human head and neck squamous carcinoma cells. Anticancer Res. 32:753–760. 2012.PubMed/NCBI
12.	Separovic D, Breen P, Joseph N, Bielawski J, Pierce JS, Van Buren E and Gudz TI: siRNA-mediated down-regulation of ceramide synthase 1 leads to apoptotic resistance in human head and neck squamous carcinoma cells after photodynamic therapy. Anticancer Res. 32:2479–2485. 2012.
13.	Gencer EB, Ural AU, Avcu F and Baran Y: A novel mechanism of dasatinib-induced apoptosis in chronic myeloid leukemia; ceramide synthase and ceramide clearance genes. Ann Hematol. 90:1265–1275. 2011. View Article : Google Scholar : PubMed/NCBI
14.	Khurana D, Martin EA, Kasperbauer JL, O’Malley BW Jr, Salomao DR, Chen L and Strome SE: Characterization of a spontaneously arising murine squamous cell carcinoma (SCC VII) as a prerequisite for head and neck cancer immunotherapy. Head Neck. 23:899–906. 2001. View Article : Google Scholar : PubMed/NCBI
15.	Suit HD, Sedlacek RS, Silver G and Dosoretz D: Pentobarbital anesthesia and the response of tumor and normal tissue in the C3Hf/sed mouse to radiation. Radiat Res. 104:47–65. 1985. View Article : Google Scholar : PubMed/NCBI
16.	Separovic D, Semaan L, Tarca AL, Awad Maitah MY, Hanada K, Bielawski J, Villani M and Luberto C: Suppression of sphingomyelin synthase 1 by small interference RNA is associated with enhanced ceramide production and apoptosis after photodamage. Exp Cell Res. 314:1860–1868. 2008. View Article : Google Scholar : PubMed/NCBI
17.	Bai A, Szulc ZM, Bielawski J, Mayroo N, Liu X, Norris J, Hannun YA and Bielawska A: Synthesis and bioevaluation of omega-N-amino analogs of B13. Bioorg Med Chem. 17:1840–1848. 2009. View Article : Google Scholar : PubMed/NCBI
18.	Dolgachev V, Farooqui MS, Kulaeva OI, Tainsky MA, Nagy B, Hanada K and Separovic D: De novo ceramide accumulation due to inhibition of its conversion to complex sphingolipids in apoptotic photosensitized cells. J Biol Chem. 279:23238–23249. 2004. View Article : Google Scholar : PubMed/NCBI
19.	Dolgachev V, Nagy B, Taffe B, Hanada K and Separovic D: Reactive oxygen species generation is independent of de novo sphingolipids in apoptotic photosensitized cells. Exp Cell Res. 288:425–436. 2003. View Article : Google Scholar : PubMed/NCBI
20.	Separovic D, Saad ZH, Edwin EA, Bielawski J, Pierce JS, Van Buren E and Bielawska A: C16-ceramide analog combined with Pc 4 photodynamic therapy evokes enhanced total ceramide accumulation, promotion of DEVDase activation in the absence of apoptosis, and augmented overall cell killing. J Lipids. 2011:1–9. 2011. View Article : Google Scholar
21.	Separovic D, Mann KJ and Oleinick NL: Association of ceramide accumulation with photodynamic treatment-induced cell death. Photochem Photobiol. 68:101–109. 1998. View Article : Google Scholar : PubMed/NCBI
22.	Agarwal ML, Clay ME, Harvey EJ, Evans HH, Antunez AR and Oleinick NL: Photodynamic therapy induces rapid cell death by apoptosis in L5178Y mouse lymphoma cells. Cancer Res. 51:5993–5996. 1991.PubMed/NCBI
23.	Wispriyono B, Schmelz E, Pelayo H, Hanada K and Separovic D: A role for the de novo sphingolipids in apoptosis of photosensitized cells. Exp Cell Res. 279:153–165. 2002. View Article : Google Scholar : PubMed/NCBI
24.	Nam S, Williams A, Vultur A, List A, Bhalla K, Smith D, Lee FY and Jove R: Dasatinib (BMS-354825) inhibits Stat5 signaling associated with apoptosis in chronic myelogenous leukemia cells. Mol Cancer Ther. 6:1400–1405. 2007. View Article : Google Scholar : PubMed/NCBI
25.	Lin YC, Wu MH, Wei TT, Chuang SH, Chen KF, Cheng AL and Chen CC: Degradation of epidermal growth factor receptor mediates dasatinib-induced apoptosis in head and neck squamous cell carcinoma cells. Neoplasia. 14:463–475. 2012.PubMed/NCBI
26.	Xue T, Luo P, Zhu H, Zhao Y, Wu H, Gai R, Wu Y, Yang B, Yang X and He Q: Oxidative stress is involved in Dasatinib-induced apoptosis in rat primary hepatocytes. Toxicol Appl Pharmacol. 261:280–291. 2012. View Article : Google Scholar : PubMed/NCBI
27.	Rodriguez-Enfedaque A, Delmas E, Guillaume A, Gaumer S, Mignotte B, Vayssiere JL and Renaud F: zVAD-fmk upregulates caspase-9 cleavage and activity in etoposide-induced cell death of mouse embryonic fibroblasts. Biochim Biophys Acta. 1823:1343–1352. 2012. View Article : Google Scholar : PubMed/NCBI
28.	Mahdy AE, Cheng JC, Li J, Elojeimy S, Meacham WD, Turner LS, Bai A, Gault CR, McPherson AS, Garcia N, Beckham TH, Saad A, Bielawska A, Bielawski J, Hannun YA, Keane TE, Taha MI, Hammouda HM, Norris JS and Liu X: Acid ceramidase upregulation in prostate cancer cells confers resistance to radiation: AC inhibition, a potential radiosensitizer. Mol Ther. 17:430–438. 2009. View Article : Google Scholar : PubMed/NCBI
29.	Nagy B, Chiu S-M and Separovic D: Fumonisin B1 does not prevent apoptosis in A431 human epidermoid carcinoma cells after photosensitization with phthalocyanine 4. J Photochem Photobiol B. 57:132–141. 2000. View Article : Google Scholar : PubMed/NCBI
30.	Xue LY, Chiu SM and Oleinick NL: Photodynamic therapy-induced death of MCF-7 human breast cancer cells: a role for caspase-3 in the late steps of apoptosis but not for the critical lethal event. Exp Cell Res. 263:145–155. 2001. View Article : Google Scholar
31.	Separovic D, Joseph N, Breen P, Bielawski J, Pierce JS, van Buren E, Bhatti G, Saad ZH, Bai A and Bielawska A: Combining anticancer agents photodynamic therapy and LCL85 leads to distinct changes in the sphingolipid profile, autophagy, caspase-3 activation in the absence of cell death, and long-term sensitization. Biochem Biophys Res Commun. 409:372–377. 2011. View Article : Google Scholar
32.	Min J, Mesika A, Sivaguru M, Van Veldhoven PP, Alexander H, Futerman AH and Alexander S: (Dihydro)ceramide synthase 1 regulated sensitivity to cisplatin is associated with the activation of p38 mitogen-activated protein kinase and is abrogated by sphingosine kinase 1. Mol Cancer Res. 5:801–812. 2007. View Article : Google Scholar : PubMed/NCBI
33.	Dumka D, Puri P, Carayol N, Lumby C, Balachandran H, Schuster K, Verma AK, Terada LS, Platanias LC and Parmar S: Activation of the p38 Map kinase pathway is essential for the antileukemic effects of dasatinib. Leuk Lymphoma. 50:2017–2029. 2009. View Article : Google Scholar : PubMed/NCBI
34.	Whitacre CM, Feyes DK, Satoh T, Grossmann J, Mulvihill JW, Mukhtar H and Oleinick NL: Photodynamic therapy with the phthalocyanine photosensitizer Pc 4 of SW480 human colon cancer xenografts in athymic mice. Clin Cancer Res. 6:2021–2027. 2000.PubMed/NCBI
35.	Koybasi S, Senkal CE, Sundararaj K, Spassieva S, Bielawski J, Osta W, Day TA, Jiang JC, Jazwinski SM, Hannun YA, Obeid LM and Ogretmen B: Defects in cell growth regulation by C18:0-ceramide and longevity assurance gene 1 in human head and neck squamous cell carcinomas. J Biol Chem. 279:44311–44319. 2004. View Article : Google Scholar : PubMed/NCBI
36.	Senkal CE, Ponnusamy S, Bielawski J, Hannun YA and Ogretmen B: Antiapoptotic roles of ceramide-synthase-6-generated C16-ceramide via selective regulation of the ATF6/CHOP arm of ER-stress-response pathways. FASEB J. 24:296–308. 2010. View Article : Google Scholar : PubMed/NCBI
37.	Mayer LD and Janoff AS: Optimizing combination chemotherapy by controlling drug ratios. Mol Interv. 7:216–223. 2007. View Article : Google Scholar : PubMed/NCBI