In vitro and in vivo therapeutic approach for a small cell carcinoma of the ovary hypercalcaemic type using a SCCOHT-1 cellular model

Orphanet Journal of Rare Diseases

Ralf Hass¹  Juliane von der Ohe¹  Tjoung-Won Park-Simon¹  Peter Hillemanns¹  Finn Rauprich¹  Anna Otte¹ 

[1] Biochemistry and Tumor Biology Laboratory, Department of Gynecology and Obstetrics Medical, University Hannover, Carl-Neuberg-Str. 1, Hannover, D – 30625, Germany

关键词: Chemotherapy;    Tumor growth;    Ovarian cancer;    SCCOHT;   

Others  :  1150005
DOI  :  10.1186/s13023-014-0126-4

 received in 2014-04-01, accepted in 2014-07-23,  发布年份 2014

【 摘 要 】

Background

The small cell ovarian carcinoma of the hypercalcemic type (SCCOHT) which preferably affects young women during regenerative age represents a rare and aggressive form of ovarian tumors with poor prognosis and lacks an efficient therapy.

Methods and results

In vitro chemotherapy testing in a fluorescence assay using a recently developed cellular model from a recurrent SCCOHT revealed sensitivity for certain epothilones, methotrexate and topotecan whereas little if any cytotoxicity was observed with other chemotherapeutics including platin-based compounds. In particular, epothilone B demonstrated a high sensitivity in contrast to ixabepilone with only little detectable effects. Western blot and cell cycle analysis revealed that the epothilone B sensitivity was associated with increased Ser¹⁵ phosphorylation of p53, a significant G₁ and G₂ cell cycle accumulation and subsequent cell death in subG₁ phase. Moreover, tubulinβ3 expression in SMARCA4/BRG1-defective SCCOHT-1 in contrast to other ovarian cancer cells was also affected during chemotherapy treatment. Increased extracellular Ca²⁺ levels further enhanced the epothilone B cytotoxicity in SCCOHT-1 cells. These in vitro effects were also confirmed in vivo in NOD/scid mouse xenografts demonstrating an attenuated tumor growth in epothilone B / Ca²⁺-treated mice. After 4d of subsequent treatment, the tumor sizes were reduced by about 90% as compared to continuously growing control tumors. In parallel, a hypercalcemia in control tumor-carrying mice was reverted to normal serum Ca²⁺ levels after epothilone B / Ca²⁺ therapy.

Conclusions

Taken together, these data demonstrated anti-tumorigenic effects of epothilone B / Ca²⁺ in SCCOHT providing a focused therapeutic approach against this rare disease and arising recurrent tumors.

【 授权许可】

   
2014 Otte et al. ; Licensee Biomedcentral Ltd

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Ulbright TM, Roth LM, Stehman FB, Talerman A, Senekjian EK: Poorly differentiated (small cell) carcinoma of the ovary in young women: evidence supporting a germ cell origin. Hum Pathol 1987, 18:175-184.
• [2]Young RH, Oliva E, Scully RE: Small cell carcinoma of the hypercalcemic type in the ovary. Gynecol Oncol 1995, 57:7-8.
• [3]Aguirre P, Thor AD, Scully RE: Ovarian small cell carcinoma. Histogenetic considerations based on immunohistochemical and other findings. Am J Clin Pathol 1989, 92:140-149.
• [4]Walt H, Hornung R, Fink D, Dobler-Girdziunaite D, Stallmach T, Spycher MA, Maly F, Haller U, Burki N: Hypercalcemic-type of small cell carcinoma of the ovary: characterization of a new tumor line. Anticancer Res 2001, 21:3253-3259.
• [5]McCluggage WG, Oliva E, Connolly LE, McBride HA, Young RH: An immunohistochemical analysis of ovarian small cell carcinoma of hypercalcemic type. Int J Gynecol Pathol 2004, 23:330-336.
• [6]Upchurch KS, Parker LM, Scully RE, Krane SM: Differential cyclic AMP responses to calcitonin among human ovarian carcinoma cell lines: a calcitonin-responsive line derived from a rare tumor type. J Bone Miner Res 1986, 1:299-304.
• [7]Dickersin GR, Kline IW, Scully RE: Small cell carcinoma of the ovary with hypercalcemia: a report of eleven cases. Cancer 1982, 49:188-197.
• [8]Eichhorn JH, Bell DA, Young RH, Swymer CM, Flotte TJ, Preffer RI, Scully RE: DNA content and proliferative activity in ovarian small cell carcinomas of the hypercalcemic type. Implications for diagnosis, prognosis, and histogenesis. Am J Clin Pathol 1992, 98:579-586.
• [9]Jelinic P, Mueller JJ, Olvera N, Dao F, Scott SN, Shah R, Gao J, Schultz N, Gonen M, Soslow RA, Berger MF, Levine DA: Recurrent SMARCA4 mutations in small cell carcinoma of the ovary. Nat Genet 2014, 46:424-426.
• [10]Witkowski L, Carrot-Zhang J, Albrecht S, Fahiminiya S, Hamel N, Tomiak E, Grynspan D, Saloustros E, Nadaf J, Rivera B, Gilpin C, Castellsague E, Silva-Smith R, Plourde F, Wu M, Saskin A, Arseneault M, Karabakhtsian RG, Reilly EA, Ueland FR, Margiolaki A, Pavlakis K, Castellino SM, Lamovec J, Mackay HJ, Roth LM, Ulbright TM, Bender TA, Georgoulias V, Longy M, et al.: Germline and somatic SMARCA4 mutations characterize small cell carcinoma of the ovary, hypercalcemic type. Nat Genet 2014, 46:438-443.
• [11]Ramos P, Karnezis AN, Craig DW, Sekulic A, Russell ML, Hendricks WP, Corneveaux JJ, Barrett MT, Shumansky K, Yang Y, Shah SP, Prentice LM, Marra MA, Kiefer J, Zismann VL, McEachron TA, Salhia B, Prat J, D'Angelo E, Clarke BA, Pressey JG, Farley JH, Anthony SP, Roden RB, Cunliffe HE, Huntsman DG, Trent JM: Small cell carcinoma of the ovary, hypercalcemic type, displays frequent inactivating germline and somatic mutations in SMARCA4. Nat Genet 2014, 46:427-429.
• [12]Ungefroren H, Sebens S, Seidl D, Lehnert H, Hass R: Interaction of tumor cells with the microenvironment. Cell Commun Signal 2011, 9:18.
• [13]Hass R, Otte A: Mesenchymal stem cells as all-round supporters in a normal and neoplastic microenvironment. Cell Commun Signal 2012, 10:26.
• [14]Otte A, Gohring G, Steinemann D, Schlegelberger B, Groos S, Langer F, Kreipe HH, Schambach A, Neumann T, Hillemanns P, Park-Simon TW, Hass R: A tumor-derived population (SCCOHT-1) as cellular model for a small cell ovarian carcinoma of the hypercalcemic type. Int J Oncol 2012, 41:765-775.
• [15]Bernhardt G, Muller R, Gust R, Reile H, Keller C, Spruss T, Schonenberger H: Dichloro-[1-(hydroxyphenyl)-2-phenylethylenediamine]platinum(II) complexes: testing on the human ovarian cancer cell lines NIH: OVCAR3 and SK OV 3. Arch Pharm 1992, 325:93-99.
• [16]Bertram C, Hass R: Cellular senescence of human mammary epithelial cells (HMEC) is associated with an altered MMP-7/HB-EGF signaling and increased formation of elastin-like structures. Mech Ageing Dev 2009, 130:657-669.
• [17]Tomayko MM, Reynolds CP: Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol 1989, 24:148-154.
• [18]Shrimali RK, Correa PD, Reed NS: Dose-dense and dose-intense chemotherapy for small cell ovarian cancer: 2 cases and review of literature. Med Oncol (Northwood London England) 2012, 28:766-770.
• [19]Harrison ML, Hoskins P, du Bois A, Quinn M, Rustin GJ, Ledermann JA, Baron-Hay S, Friedlander ML: Small cell of the ovary, hypercalcemic type – analysis of combined experience and recommendation for management. A GCIG Study Gynecol Oncol 2006, 100:233-238.
• [20]Benrubi GI, Pitel P, Lammert N: Small cell carcinoma of the ovary with hypercalcemia responsive to sequencing chemotherapy. South Med J 1993, 86:247-248.
• [21]Reed WC: Small cell carcinoma of the ovary with hypercalcemia: report of a case of survival without recurrence 5 years after surgery and chemotherapy. Gynecol Oncol 1995, 56:452-455.
• [22]Dykgraaf RH, de Jong D, van Veen M, Ewing-Graham PC, Helmerhorst TJ, van der Burg ME: Clinical management of ovarian small-cell carcinoma of the hypercalcemic type: a proposal for conservative surgery in an advanced stage of disease. Int J Gynecol Cancer 2009, 19:348-353.
• [23]Barondeau J, Rodgers M, Braun L, Azarow K, Forouhar M, Faucette K: Small cell ovarian carcinoma: a rare, aggressive tumor masquerading as constipation in a teenager with a fatal outcome. J Pediatr Hematol Oncol 2010, 32:e139-e141.
• [24]Gamwell LF, Gambaro K, Merziotis M, Crane C, Arcand SL, Bourada V, Davis C, Squire JA, Huntsman DG, Tonin PN, Vanderhyden BC: Small cell ovarian carcinoma: genomic stability and responsiveness to therapeutics. Orphanet J Rare Dis 2010, 8:33.
• [25]Stetler RA, Gao Y, Zhang L, Weng Z, Zhang F, Hu X, Wang S, Vosler P, Cao G, Sun D, Graham SH, Chen J: Phosphorylation of HSP27 by protein kinase D is essential for mediating neuroprotection against ischemic neuronal injury. J Neurosci 2012, 32:2667-2682.
• [26]Bertram C, Hass R: Cellular responses to reactive oxygen species-induced DNA damage and aging. Biol Chem 2008, 389:211-220.
• [27]Damia G, Filiberti L, Vikhanskaya F, Carrassa L, Taya Y, D'Incalci M, Broggini M: Cisplatinum and taxol induce different patterns of p53 phosphorylation. Neoplasia (New York, NY 2001, 3:10-16.
• [28]Ioffe ML, White E, Nelson DA, Dvorzhinski D, DiPaola RS: Epothilone induced cytotoxicity is dependent on p53 status in prostate cells. Prostate 2004, 61:243-247.
• [29]Khrapunovich-Baine M, Menon V, Yang CP, Northcote PT, Miller JH, Angeletti RH, Fiser A, Horwitz SB, Xiao H: Hallmarks of molecular action of microtubule stabilizing agents: effects of epothilone B, ixabepilone, peloruside A, and laulimalide on microtubule conformation. J Biol Chem 2011, 286:11765-11778.
• [30]Kavallaris M: Microtubules and resistance to tubulin-binding agents. Nat Rev 2010, 10:194-204.
• [31]Seve P, Dumontet C: Is class III beta-tubulin a predictive factor in patients receiving tubulin-binding agents? Lancet Oncol 2008, 9:168-175.
• [32]Otte A, Rauprich F, von der Ohe J, Hillemanns P, Hass R: Interference of Ca2+ with the proliferation of SCCOHT-1 and ovarian adenocarcinoma cells. Int J Oncol 2014, 45:1151-1158.