Ursolic acid from Trailliaedoxa gracilis induces apoptosis in medullary thyroid carcinoma cells

Aguiriano‑Moser,Victor; Svejda,Bernhard; Li,Zeng‑Xia; Sturm,Sonja; Stuppner,Hermann; Ingolic,Elisabeth; Höger,Harald; Siegl,Veronika; Meier-Allard,Nathalie; Sadjak,Anton; Pfragner,Roswitha

doi:10.3892/mmr.2015.4053

Ursolic acid from Trailliaedoxa gracilis induces apoptosis in medullary thyroid carcinoma cells

Authors:
- Victor Aguiriano‑Moser
- Bernhard Svejda
- Zeng‑Xia Li
- Sonja Sturm
- Hermann Stuppner
- Elisabeth Ingolic
- Harald Höger
- Veronika Siegl
- Nathalie Meier-Allard
- Anton Sadjak
- Roswitha Pfragner
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Affiliations: Department of Pathophysiology and Immunology, Center of Molecular Medicine, Medical University of Graz, Graz A‑8010, Austria, Department of Pharmacognosy, Institute of Pharmacy, Center of Molecular Biosciences, Leopold Franzens University of Innsbruck, Innsbruck A‑6010, Austria, Core Unit of Biomedical Research, Division of Laboratory Animal Science and Genetics, Medical University of Vienna, Himberg A‑2325, Austria, Research Institute for Electron Microscopy and Fine Structure Research, University of Technology Graz, Graz A‑8010, Austria
Published online on: July 7, 2015 https://doi.org/10.3892/mmr.2015.4053
Pages: 5003-5011
Copyright: © Aguiriano‑Moser et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Medullary thyroid carcinoma (MTC) originates from the C‑cells of the thyroid and is not sensitive to radiation or chemotherapy. Therefore, surgical removal of the tumor tissue in its entirety is the only curative treatment for MTC. The present study aimed to examine the potential mechanisms of action of extracts of Trailliaedoxa gracilis (TG; WW Smith & Forrest), a plant from the province of Sichuan, China, and of ursolic acid (UA), a pentacyclic triterpen present in TG, on the MTC‑SK MTC cell line. A total of 13 TG fractions and UA were examined in vitro for their effects on cell morphology, cell number, proliferation and rates of apoptosis. Reverse transcription‑quantitative polymerase chain reaction of nuclear factor‑κB essential modifier (NEMO) was performed to delineate the role of the apoptotic pathway following treatment with UA. TG and UA were examined in vivo in xenotransplanted MTC‑bearing severe combined immunodeficient mice. The TG fractions exhibited antiproliferative effects, with inhibition of mitochondrial activity in the tumor cells at concentrations, which caused no impairment of the normal control cells. The apoptotic rates of the MTC‑SK cells treated with the TG fractions and UA were determined, in which no marked tumor inhibition was observed in the treated MTC‑mice, and no change in the expression of NEMO was detected in the treated MTC‑SK cells. The observation of early‑onset activation of caspase 8 suggested that the responsible factor was linked to NEMO, an anti‑apoptotic protein. However, no differences in the mRNA transcription levels of NEMO were detected in MTC‑SK cells treated with UA, suggesting that this protein was not associated with the signal transducer and activator of transcription 3 pathway.

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1	Pfragner R, Höfler H, Behmel A, Ingolic E and Walser V: Establishment and characterization of continuous cell line MTC-SK derived from a human medullary thyroid carcinoma. Cancer Res. 50:4160–4166. 1990.PubMed/NCBI
2	Herold G: Innere Medizin. Herold; Köln: pp. 743–745. 2012, In German.
3	Nicolini V, Cassinelli G, Cuccuru G, Bongarzone I, Petrangolini G, Tortoreto M, Mondellini P, Casalin P, Favini E, Zaffaroni N, Zunino F and Lanzi C: Interplay between Ret and Fap-1 regulates CD95-mediated apoptosis in medullary thyroid cancer cells. Biochem Pharmacol. 82:778–788. 2011. View Article : Google Scholar : PubMed/NCBI
4	Schulten HJ, Al-Maghrabi J, Al-Gahmdi K, Salama S, Al-Muhayawi S, Chaudhary A, Hamour O, Abuzenadah A, Gari M and Al-Qhatani M: Mutational screening of RET, HRAS, KRAS, NRAS, BRAF, AKT1 and CTNNB1 in medullary thyroid carcinoma. Anticancer Res. 31:4179–4183. 2011.PubMed/NCBI
5	Li ZX, Sturm S, Stuppner H, Schraml E, Aguiriano-Moser V, Siegl V and Pfragner R: The dichloromethane fraction of Stemona tuberosa Lour inhibits tumor cell growth and induces apoptosis of human medullary thyroid carcinoma cells. Biologics: Targets and Therapy. 1:455–463. 2007.
6	Li Z, Sturm S, Svejda B, Höger H, Schraml E, Ingolic E, Siegl V, Stuppner H and Pfragner R: Anticancer activity of novel extracts from Cautleya gracilis (Smith) Dandy: apoptosis in human medullary thyroid carcinoma cells. Anticancer Res. 28(5A): 2705–2713. 2008.PubMed/NCBI
7	Svejda B, Aguiriano-Moser V, Sturm S, Höger H, Ingolic E, Siegl V, Stuppner H and Pfragner R: Anticancer antivity of novel plant extracts from Trailliaedoxa gracilis (W.W. Smith & Forrest) in human carcinoid KRJ-I cells. Anticancer Res. 30:55–64. 2010.PubMed/NCBI
8	Rinner B, Siegl V, Pürstner P, Efferth T, Brem B, Greger H and Pfragner R: Activity of novel plant extracts against medullary thyroid carcinoma cells. Anticancer Res. 24(2A): 495–500. 2004.PubMed/NCBI
9	Rinner B, Li ZX, Haas H, Siegl V, Sturm S, Stuppner H and Pfragner R: Antiproliferative and pro-apoptotic effects of Uncaria tormentosa in human medullary thyroid carcinoma cells. Anticancer Res. 29:4519–28. 2009.PubMed/NCBI
10	Tappeiner J: Phytochemische Untersuchungen zu Trailliaedoxa gracilis,Diplomarbeit zur Erlangung des akademischen Grades Magistra der Pharmazie an der Leopold Franzens Universität Innsbruck. pp. 15–50. 2007, In German.
11	Handel-Mazzetti H: Naturbilder aus Südwest-China Erlebnisse und Eindrücke eines österreichischen Forschers während des Weltkrieges. Österreichischer Bundesverlag; Wien-Leipzig: pp. 3801927, In German.
12	Wu Hao, Zeng Liangqing and Gong Xun: Chromosome number of Traillaedoxa gracilis (Rubiaceae) endemic to jinsha river valley. Acta Botanica Yunnanica. 32:407–408. 2010.
13	Pathak AK, Bhutani M and Nair AS: Ursolic acid inhibits STAT3 activation pathway leading to supression of proliferation and chemosensitization of human myeloma cells. Mol Canc Res. 5:943–955. 2007. View Article : Google Scholar
14	Liobikas J, Majiene D, Trumbeckaite S, Kursvietiene K, Masteikova R, Kopustinkinene DM, Savickas A and Bernatoniene J: Uncoupling and antioxidant effects of ursolic acid in isolated rat heart mitochondria. J Nat Prod. 74:1640–1644. 2011. View Article : Google Scholar : PubMed/NCBI
15	Wu WY, Li J, Wu ZS, Zhang CL and Meng XL: STAT3 activation in monocytes accelerates liver cancer progression. BMC Cancer. 11:5062011. View Article : Google Scholar : PubMed/NCBI
16	Liu Q, Liu S, Shi Y, Li H, Hao J, Xing L, Cao Y and Duan H: Suppressors of cytokine signaling inhibit tubular, epithelial cell-myofibroblast transdifferentiation. Am J Nephrol. 34:142–151. 2011. View Article : Google Scholar
17	Li X, Massa PE, Hanidu A, Peet GW, Aro P, Savitt A, Mische S, Li J and Marcu KB: IKKalpha, IKKbeta and NEMO/IKKgamma are each required for the NF-kappa B-mediated inflamatory response program. J Biol Chem. 277:45129–45140. 2002. View Article : Google Scholar : PubMed/NCBI
18	Miyamoto S: Nuclear initiated NF-κB signaling: NEMO and ATM take center stage. Rev Cell Res. 21:116–130. 2011. View Article : Google Scholar
19	Rudolph D, Yeh WC, Wakeham A, Rudolph B, Nallainathan D, Potter J, Elia AJ and Mak TW: Severe liver degeneration and lack of NF-kappaB activation in NEMI/IKKgamma-deificient mice. Genes De. 14:854–862. 2000.
20	Legarda-Addison D, Hase H, O'Donnell MA and Ting AT: NEMO/IKKgamma regulates an early NF-kappaB-independent cell death checkpoint during TNF signaling. Cell Death Differ. 16:1297–1288. 2009. View Article : Google Scholar
21	Kojima H, Sasaki T, Ishitani T, Iemura S, Zhao H, Kaneko S, Kunimoto H, Natsume T, Matsumoto K and Kakajima K: STAT3 regulates nemo-like kinase by mediating its interaction with IL-6-stimulated TGFbeta-activated kinase 1 for STAT3 Ser 727 phosphorilation. Proc Natl Acad Sci USA. 102:4524–4529. 2005. View Article : Google Scholar
22	Wolf C, Lederer K, Pfragner R, Schauenstein K, Ingolic E and Siegl V: Biocompatibility of ultra-high molecular weight polyethylene (UHMW-PE) stablized with alpha-tocopherol used for joint endoprotheses assessed in vitro. J Mater Sci Mater Med. 18:1247–1252. 2007. View Article : Google Scholar : PubMed/NCBI
23	Pfragner R, Behmel A, Ingolic E and Wirnsberger GH: Culture of human neuroendocrine tumor cells. Culture of Specialized Cells: Culture of human tumor cells. Pfragner R and Freshney RI: John Wiley & Sons, Inc; Hoboken, New Jersey: pp. 373–403. 2004
24	Pfaffl MW: A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29:e452001. View Article : Google Scholar : PubMed/NCBI
25	Aggarwal BB and Shishodia S: Molecular targets of dietary agents for prevention and therapy of cancer. Biochem Pharmacol. 71:1397–1421. 2006. View Article : Google Scholar : PubMed/NCBI
26	Pfragner R, Wirnsberger G, Behmel A, Wolf G, Passath GA, Ingolic E and Adamiker D: New continuous cell line from human medullary carcinoma: SINJ. Phenotypic analysis and in vivo carcinogenesis. Int J Oncol. 2:831–836. 1993.PubMed/NCBI
27	Pfragner R, Wirnsberger G, Ingolic E and Niederle B: Medullary thyroid carcinomas in cell culture - models for future therapies. Wien Klin Wochenschr. 114:279–283. 2002.PubMed/NCBI
28	Pfragner R, Wirnsberger G, Niederle B, Behmel A, Rinner I, Mandl A, Wawrina F, Luo JS, Adamiker D, Höger H, Ingolic E and Schauenstein K: Establishment of a continuous cell line from a human carcinoid of the small intestine (KRJ-I): Characterization and effects of 5-azacytidine on proliferation. Int J Onc. 8:513–520. 1996.
29	Kimhi Y, Palfrey C, Spector I, Barak Y and Littauer UZ: Maturation of neuroblastoma cells in the presence of dimethyl-sulfoxide. Proc Nat Acad Sci USA. 73:462–466. 1976. View Article : Google Scholar
30	Mitaka T, Norioka K, Nakamura T and Mochizukio Y: Effects of mitogens and co-mitogens on the formation of small-cell colonies in primary cultures of rat hepatocytes. J Cell Physiol. 157:461–468. 1993. View Article : Google Scholar : PubMed/NCBI
31	Rudland PS: Use of peanut lectin and rat mammary stem cell lines to identify a cellular differentiation pathway for the alveolar cell in the rat mammary gland. J Cell Physiol. 152:157–168. 1992. View Article : Google Scholar
32	Da Violante G, Zerrouk N, Richard I, Provot G, Chaumeil JC and Arnaud P: Evaluation of the cytotoxicity effect of dimethyl sulfoxide (DMSO) on Caco2/TC7 colon tumor cell cultures. Biol Pharm Bull. 25:1600–1603. 2002. View Article : Google Scholar : PubMed/NCBI
33	Boatright KM and Salvesen GS: Mechanisms of caspase activation. Current Op in Cell Biol. 15:725–731. 2003. View Article : Google Scholar
34	Fusco F, Mercadante V, Miano MG and Ursini MV: Multiple regulatory regions and tissue-specific transcription initiation mediate the expression of NEMO/IKKγ gene. Gene. 383:99–107. 2006. View Article : Google Scholar : PubMed/NCBI
35	Shishodia S, Majumdar S, Banerjee S and Aggarwaal BB: Ursolic acid inhibits nuclear factor-κB activation induced by carcinogenic agents through suppression of IκBa kinase and p65 phosphorylation: correlation with down-regulation of cyclo-oxygenase 2, matrix metalloproteinase 9 and cyclin D11. Cancer Res. 63:4375–4383. 2003.PubMed/NCBI
36	Chen ZJ: Ubiquitination in signaling to and activation of IKK. Immunol Rev. 246:95–106. 2012. View Article : Google Scholar : PubMed/NCBI
37	Liu S, Lv J, Han L, Ichikawa T, Wang W, Li S, Wang XL, Tang D and Cui T: A pro-inflammatory role of deubiquitinating enzyme cylindromatosis (CYLD) in vascular smooth muscle cells. Biochem Biophys Res Commun. 420:78–83. 2012. View Article : Google Scholar : PubMed/NCBI
38	Wang Q, Yuan L, Liu Z, Jiang X and Lu J: Expression of A20 is reduced in pancreatic cancer tissues. J Mol Histol. 43:319–325. 2012. View Article : Google Scholar : PubMed/NCBI
39	Schmukle AC and Walczak H: No one can whistle a symphony alone-how different ubiquitin linkages cooperate to orchestrate NF-κB activity. J Cell Sci. 125:549–559. 2012. View Article : Google Scholar : PubMed/NCBI