Djanan VEJSELOVA, Hatice Mehtap KUTLU, Gökhan KUŞ, Selda KABADERE, Ruhi UYAR

Cytotoxic and apoptotic effects of ceranib-2 offering potential for a new antineoplastic agent in the treatment of cancer cells

Fibrocarcinomas are malignant tumours that originate in mesenchymal cells. The tumours typically form in the presence of connective tissue and are of key consideration when assessing research priorities for future healthcare management. Ceramidase inhibitors, such as ceranib-2, have demonstrated capacity to interfere with cellular DNA functionality, initiating apoptosis in many cancer cell lines. The enzyme ceramidase can regulate cellular levels of sphingosine and sphingosine-1-phosphate by controlling hydrolysis of ceramide. The present study investigates the antigrowth effects of ceranib-2 on mouse embryonic fibroblast cells (NIH/3T3 normal cell line) and rat embryonic fibroblast cells (5RP7 cancer cell line). Research was conducted using colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, confocal microscopy, and transmission electron microscopy. The results indicate that in vitro fibrocarcinomas have the potential to undergo apoptotic death when influenced by the antigrowth behaviour of ceranib-2. This is true even of cells exposed only to minimal doses of ceranib-2. Indicators of apoptotic death, specifically presentation of a pyknotic nucleus, were observed by confocal micrographs in treated cells double-stained with acridine orange and Alexa Fluor-488 Phalloidin. It is theorised that ceranib-2 will function in accordance with the antiproliferative activity of other acid ceramidases, ultimately resulting in the molecule's classification as a new antitumour agent.

Anahtar Kelimeler:

Ceramidase inhibitor, ceranib-2, cancer treatment, cytotoxicity, apoptosis

Cytotoxic and apoptotic effects of ceranib-2 offering potential for a new antineoplastic agent in the treatment of cancer cells

Keywords:

Ceramidase inhibitor, ceranib-2, cancer treatment, cytotoxicity, apoptosis,

PDF

___

Akao Y, Banno Y, Nakagawa Y, Hasegawa N, Kim TJ, Murate T, Igarashi Y, Nzawa Y (2006). High expression of sphingosine kinase 1 and SIP receptors in chemotherapy resistant prostate cancer PC3 cells and their camptothecin induced upregulation. Biochem Bioph Res Co 342: 1284–1290.
Beckham TH, Elojemy S, Cheng JC, Turner LS, Hoffman S, Norris JS, Liu X (2010). Targeting sphingolipid metabolism in head and neck cancer: rational therapeutic potentials. Expert Opin Ther Targets 14: 529–539.
Canals D, Perry DM, Jenkins RW, Hannun YA (2011). Drug targeting of sphingolipid metabolism: sphingomyelinases and ceramidases. Br J Pharmacol 163: 694–712.
Chalfant CE, Spiegel S (2005). Sphingosine 1-phosphate and ceramide 1-phosphate: expanding roles in cell signaling. J Cell Sci 118: 4605–4622.
Chen LC, Lin CF, Chang WT, Huang WC, Teng CF, Lin YS (2008). Ceramide induces p38 MAPK and JNK activation through a mechanism involving a thioredoxin-interacting protein- mediated pathway. Blood 111: 4365–4374.
Draper JM, Xia Z, Smith RA, Zhuang Y, Wang W, Smith CD (2011). Discovery and evaluation of inhibitors of human ceramidase. Mol Cancer Ther 10: 2052–2061.
Hofmann K, Tomiuk S, Wolff G, Stoffel W (2000). Cloning and characterization of the mammalian brain-specific, Mg2- dependent neutral sphingomyelinase. P Natl Acad Sci U S A 97: 5895–5900.
Kawamori T, Kaneshiro T, Okumura M, Maalouf S, Uflaceker A, Bielawski J, Hannun YA, Obeid LM (2009). Role of sphingosine kinase in colon carcinogenesis. FASEB J 23: 405–414.
Koch J, Gartner S, Li CM, Quintern LE, Bernardo K, Levran O, Schnabel D, Desnick RJ, Schuchman EH, Sandhoff K (1996). Molecular cloning and characterization of a full-length complementary DNA encoding human acid ceramidase. Identification of the first molecular lesion causing Farber disease. J Biol Chem 271: 33110–33115.
Kohama T, Olivera A, Edsall L, Nagiec MM, Dickson R, Spiegel S (1998). Molecular cloning and functional characterization of murine sphingosine kinase. J Biol Chem 273: 23722–23728.
Liu H, Sugiura M, Nava VE, Edsall LC, Kono K, Poulton S, Milstien S, Kohama T, Spiegel S (2000). Molecular cloning and functional characterization of a novel mammalian sphingosine kinase type 2 isoform. J Biol Chem 275: 19513–19520.
Liu Y, Wada R, Yamashita T, Mi Y, Deng CX, Hobson JP, Rosenfeldt HM, Nava VE, Chae SS, Lee MJ et al. (2000). Edg-1, the G protein-coupled receptor for sphingosine-1-phosphate, is essential for vascular maturation. J Clin Invest 106: 951–961.
Nava VE, Hobson JP, Murthy S, Milstien S, Spiegel S (2002). Sphingosine kinase type 1 promotes estrogen dependent tumorigenesis of breast cancer MCF-7 cells. Exp Cell Res 281: 115–127.
Ogretmen B, Hannun YA (2004). Biologically active sphingolipids in cancer pathogenesis and treatment. Nat Rev Cancer 4: 604– 616.
Osawa Y, Uchinami H, Bielawski I, Schwabe F, Hannun YA, Brenner DA (2005). Roles of C ceramide and sphingosine 1 phosphate I regulating hepatocyte apoptosis in response to tumor necrosis factor. J Biol Chem 280: 27879–27887.
Oskouian B, Saba JD (2010). Cancer treatment strategies targeting sphingolipid metabolism. Adv Exp Med Biol 688: 185–205.
Ponnusamy S, Needham MM, Senkal CE, Shar AS, Sentelle D, Selvam SP, Salas A, Ogretmen B (2010). Sphingolipids and cancer: ceramide and sphingosine-1-phosphate in the regulation of cell death and drug resistance. Future Oncol 6: 1603–1624.
Proksch D, Klein JJ, Arenz C (2011). Potent inhibition of acid ceramidase by novel B-13 analogues. Journal of Lipids 2011: 971618.
Quintern LE, Schuchman EH, Levran O, Suchi M, Ferlinz K, Reinke H, Sandhoff K, Desnick RJ (1989). Isolation of cDNA clones encoding human acid sphingomyelinase: occurrence of alternatively processed transcripts. EMBO J 8: 2469–2473.
Reynolds CP, Maurer BJ, Kolesnick RN (2004). Ceramide synthesis and metabolism as a target for cancer therapy. Cancer Lett 206: 169–180.
Riboni L, Campanella R, Bassi R, Villani R, Gaini SM, Martinelli- Boneschi F, Viani P, Tettamanti G (2002). Ceramide levels are inversely associated with malignant progression of human glial tumors. Glia 39: 105–113.
Sarkar S, Maceyna M, Hait NC, Paugh SW, Sankala H, Milstien S, Spiegel S (2005). Sphingosine kinase-1 is required for migration, proliferation and survival of MCF-7 human breast cancer cell. FEBS Lett 579: 5313–5317.
Seelan RS, Quian C, Yokomiza A, Bostwick G, Smith DI, Liu W (2000). Human acid ceramidase is overexpressed but not mutated in prostate cancer. Gene Chromosome Canc 29:137– 146.
Senchenkow A, Litvak DA, Cabot MC (2001). Targeting ceramide metabolism-a strategy for overcoming drug resistance. J Natl Cancer I 93: 347–357.
Strelow A, Bernardo K, Adam S, Klages A (2000). Overexpression of acid ceramidase protects from tumor necrosis factor induced cell death. J Exp Med 192: 601–611.
Taha TA, Mullen TD, Obeid LM (2006). A house divided: ceramide, sphingosine, and sphingosine-1-phosphate in programmed cell death. Biochim Biophys Acta 12: 2027–2036.
Tomiuk S, Hofmann K, Nix M, Zumbansen M, Stoffel W (1998). Cloned mammalian neutral sphingomyelinase: functionsin sphingolipid signaling? Proc Natl Acad Sci U S A 95: 3638– 3643.
Xu R, Jin J, Hu W (2006). Golgi alkaline ceramidase regulates cell proliferation and survival by controlling levels of sphingosine and S1P. FASEB J 20: 1813–1825.
Xu R, Sun W, Jin J, Obeid LM, Mao C (2010). Role of alkaline ceramidases in the generation of sphingosine and its phosphate in erythrocytes. FASEB J 24: 2507–2515.