Moscatilin induces apoptosis of pancreatic cancer cells via reactive oxygen species and the JNK/SAPK pathway

Zhang,Lei; Fang,Yuan; Xu,Xue‑Feng; Jin,Da‑Yong

doi:10.3892/mmr.2017.6144

Moscatilin induces apoptosis of pancreatic cancer cells via reactive oxygen species and the JNK/SAPK pathway

Authors:
- Lei Zhang
- Yuan Fang
- Xue‑Feng Xu
- Da‑Yong Jin
View Affiliations

Affiliations: Pancreatic Cancer Group, General Surgery Department, Zhongshan Hospital, Fudan University, Shanghai 200232, P.R. China
Published online on: January 25, 2017 https://doi.org/10.3892/mmr.2017.6144
Pages: 1195-1203
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Moscatilin is a bibenzyl derivative extracted from the Dendrobium aurantiacum var. denneanum, which has traditionally been used as an immunomodulatory treatment in China. The present study was designed to determine whether moscatilin is a pro‑apoptotic agent in pancreatic cancer, and to elucidate the underlying mechanisms. The apoptotic and anti‑proliferative effects of moscatilin on pancreatic cancer cells were determined in vitro using biochemical assays, such as the MTT assay, colony formation assay, Hoechst staining and DNA fragmentation assay, and in vivo using Panc‑1 pancreatic cancer xenografts. Western blotting was also conducted to evaluate the expression levels of B‑cell lymphoma 2 (Bcl2), Bcl2‑associated X protein (Bax), Bcl2 homologous antagonist killer (Bak), caspase 3, cleaved‑caspase 3, poly (ADP‑ribose) polymerase, p‑c‑Jun N‑terminal kinase (JNK)/stress‑activated protein kinases (SAPK) and JNK/SAPK in response to moscatilin. We used DCFH‑DA to detect the production of reactive oxygen species (ROS) induced by moscatilin. The present study demonstrated that moscatilin markedly inhibited pancreatic cancer cell viability and induced cell apoptosis in a concentration‑dependent manner. Conversely, moscatilin did not affect the cell viability of human umbilical vein endothelial cells at the comparable dosage. Treatment with moscatilin suppressed clonogenicity of Panc‑1 cells in a concentration‑dependent manner. Furthermore, a decrease in Bcl2 expression, and an increase in the expression levels of Bak and Bax, was detected following treatment with moscatilin, resulting in an increase in the proapoptotic/anti‑apoptotic expression ratio (Bax/Bcl2) in Panc‑1 cells. Moscatilin also induced activation of the caspase‑dependent mitochondrial apoptotic pathway. In addition, moscatilin enhanced cellular ROS production and induced activation of JNKSAPK signaling pathway. Conversely, pretreatment with the ROS scavenger N‑acetylcysteine or the JNK/SAPK‑specific inhibitor SP600125 prevented moscatilin‑mediated reductions in cell viability. Furthermore, moscatilin inhibited tumor growth in nude mice bearing Panc‑1 cells, without apparent toxicity. In conclusion, these results demonstrated that moscatilin may induce pancreatic cell apoptosis, and therefore may be considered a potential therapeutic agent for the treatment of pancreatic cancer.

View Figures

View References

1	Rahib L, Smith BD, Aizenber R, Rosenzweig AB, Fleshman JM and Matrisian LM: Projecting cancer incidence and deaths to 2030: The unexpected burden of thyroid, liver, and pancreas cancers in the United States. Cancer Res. 74:2913–2921. 2014. View Article : Google Scholar : PubMed/NCBI
2	Wolfgang CL, Herman JM, Laheru DA, Klein AP, Erdek MA, Fishman EK and Hruban RH: Recent progress in pancreatic cancer. CA Cancer J Clin. 63:318–348. 2013. View Article : Google Scholar : PubMed/NCBI
3	Paulson AS, Cao Tran HS, Tempero MA and Lowy AM: Therapeutic advances in pancreatic cancer. Gasroenterology. 144:1316–1326. 2013. View Article : Google Scholar
4	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar : PubMed/NCBI
5	Liou GY and Storz P: Reactive oxygen species in cancer. Free Radic Res. 44:479–496. 2010. View Article : Google Scholar : PubMed/NCBI
6	Alexandre J, Hu Y, Lu W, Pelicano H and Huang P: Novel action of paclitaxel against cancer cells: Bystander effect mediated by reactive oxygen species. Cancer Res. 67:3512–3517. 2007. View Article : Google Scholar : PubMed/NCBI
7	Paduch R, Kandefer-Szerszeń M and Piersiak T: The important of release of proinflammatory cytokines, ROS, and NO in different stages of colon carcinoma growth and metastasis after treatment with cytotoxic drugs. Oncol Res. 18:419–436. 2010. View Article : Google Scholar : PubMed/NCBI
8	Groninger E, Meeuwsen-De Boer GJ, De Graaf SS, Kamps WA and De Bont ES: Vincristine induced apoptosis in acute lymphoblastic leukaemia cells: A mitochondrial controlled pathway regulated by reactive oxygen species? Int J Oncol. 21:1339–1345. 2002.PubMed/NCBI
9	Zhang X, Xu JK, Wang J, Wang NL, Kuriihara H, Kitanaka S and Yao XS: Bioactive bibenzyl derivatives and fluorenones from Dendrobium nobile. J Nat Prod. 70:24–28. 2007. View Article : Google Scholar : PubMed/NCBI
10	Chen CC, Wu LG, Ko FN and Teng CM: Antiplatelet aggregation principles of Dendrobium loddigesii. J Nat Prod. 57:1271–1274. 1994. View Article : Google Scholar : PubMed/NCBI
11	Hu JM, Chen JJ, Yu H, Zhao YX and Zhou J: Two novel bibenzyls from Dendrobium trigonopus. J Asian Nat Prod Res. 10:653–657. 2008. View Article : Google Scholar : PubMed/NCBI
12	Gong Y, Fan Y, Liu L, Wu D, Chang Z and Wang Z: Erianin induces a JNK/SAPK-dependent metabolic inhibition in human umbilical vein endothelial cells. In Vivo. 18:223–228. 2004.PubMed/NCBI
13	Gong YQ, Fan Y, Wu DZ, Yang H, Hu ZB and Wang ZT: In vivo and in vitro evaluation of erianin, a novel anti-angiogenic agent. Eur J Cancer. 40:1554–1565. 2004. View Article : Google Scholar : PubMed/NCBI
14	Pengpaeng P, Sritularak B and Chanvorachote P: Dendrofalconerol A suppresses migrating cancer cells via EMT and integrin proteins. Anticancer Res. 35:201–205. 2015.PubMed/NCBI
15	Charoenrungruang S, Chanvorachote P, Sritularak B and Pongrakhananon V: Gigantl, a bibenzyl from Dendrobium draconis, inhibits the migratory behavior of non-small cell lung cancer cells. J Nat Prod. 77:1359–1366. 2014. View Article : Google Scholar : PubMed/NCBI
16	Chen CA, Chen CC, Shen CC, Chang HH and Chen YJ: Moscatilin induces apoptosis and mitotic catastrophe in human esophageal cancer cells. J Med Food. 16:869–877. 2013. View Article : Google Scholar : PubMed/NCBI
17	Pai HC, Chang LH, Peng CY, Chang YL, Chen CC, Shen CC, Teng CM and Pan SL: Moscatilin inhibits migration and metastasis of human breast cancer MDA-MB-231 cells through inhibition of Akt and Twist signaling pathway. J Mol Med (Berl). 91:347–356. 2013. View Article : Google Scholar : PubMed/NCBI
18	Kowitdamrong A, Chanvorachote P, Sritularak B and Pongrakhananon V: Moscatilin inhibits lung cancer cell motility and invasion via suppression of endogenous reactive oxygen species. Biomed Res Int. 2013:7658942013. View Article : Google Scholar : PubMed/NCBI
19	Tsai AC, Pan SL, Liao CH, Guh JH, Wang SW, Sun HL, Liu YN, Chen CC, Shen CC, Chang YL and Teng CM: Moscatilin, a bibenzyl derivative from the India orchid Dendrobium loddigesii, suppresses tumor angiogenesis and growth in vitro and in vivo. Cancer Lett. 292:163–170. 2010. View Article : Google Scholar : PubMed/NCBI
20	Yang L, Wang Y, Zhang G, Zhang F, Zhang Z, Wang Z and Xu L: Stimulaneous quantitative and qualitative analysis of bioactive phenols in Dendrobium aurantiacum var. Denneanum by high-performance liquid chromatography coupled with mass spectrometry and diode array detection. Biomed Chromatogr. 21:687–694. 2007. View Article : Google Scholar : PubMed/NCBI
21	Marin V, Kaplanski G, Grès S, Farnarier C and Bongrand P: Endothelial cell culture: Protocol to obtain and cultivate human umbilical endothelial cells. J Immunol Methods. 254:183–190. 2001. View Article : Google Scholar : PubMed/NCBI
22	Jo GH, Kim GY, Kim WJ, Park KY and Choi YH: Sulforaphane induces apoptosis in T24 human urinary bladder cancer cells through a reactive oxygen species-mediated mitochondrial pathway: The involvement of endoplasmic reticulum stress and the Nrf2 signaling pathway. Int J Oncol. 45:1497–1506. 2014.PubMed/NCBI
23	Kello M, Drutovic D, Chripkova M, Pilatova M, Budovska M, Kulikova L, Urdzik P and Mojzis J: ROS-dependent antiproliferative effect of brassinin derivative homobrassinin in human colorectal cancer Caco2 cells. Molecules. 19:10877–10897. 2014. View Article : Google Scholar : PubMed/NCBI
24	Vriz S, Reiter S and Galliot B: Cell death: A program to regenerate. Curr Top Dev Biol. 108:121–151. 2014. View Article : Google Scholar : PubMed/NCBI
25	Li MX and Dewson G: Mitochondria and apoptosis: Emerging concepts. F1000Prime Rep. 7:422015.PubMed/NCBI
26	Solary E, Dubrez L and Eymin B: The role of apoptosis in the pathogenesis and treatment of diseases. Eur Respir J. 9:1293–1305. 1996. View Article : Google Scholar : PubMed/NCBI
27	Suliman A, Lam A, Datta R and Srivastava RK: Intracellular mechanisms of TRAIL: Apoptosis through mitochondrial-dependent and independent-pathways. Oncogene. 20:2122–2133. 2001. View Article : Google Scholar : PubMed/NCBI
28	Vaux DL, Cory S and Adams JM: Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature. 335:440–442. 1988. View Article : Google Scholar : PubMed/NCBI
29	Czabotar PE, Lessene G, Strasser A and Adams JM: Control of apoptosis by the BCL-2 protein family: Implications for physiology and therapy. Nat Rev Mol Cell Biol. 15:49–63. 2014. View Article : Google Scholar : PubMed/NCBI
30	Tong LY, Chuang CC, Wu S and Zuo L: Reactive oxygen species in redox cancer therapy. Cancer Lett. 367:18–25. 2015. View Article : Google Scholar : PubMed/NCBI
31	Surbramani R, Gonzalez E, Arumugam A, Nandy S, Gonzalez V, Medel J, Camacho F, Ortega A, Bonkoungou S, Narayan M, et al: Nimbolide inhibits pancreatic cancer growth and metastasis through ROS-mediated apoptosis and inhibition of epithelial-mesenchymal transition. Sci Rep. 6:198192016. View Article : Google Scholar : PubMed/NCBI
32	Tian X, Dai S, Sun J, Jiang S, Sui C, Meng F, Li Y, Fu L, Jiang T, Wang Y, et al: Bufalin induces mitochondrial-dependent apoptosis in pancreatic and oral cancer cells by downregulating hTERT expression via activation of the JNK/p38 pathway. Evid Based Complement Alternat Med. 2015:5462102015. View Article : Google Scholar : PubMed/NCBI
33	Lei YY, Wang WJ, Mei JH and Wang CL: Mitogen-activated protein kinase signal transduction in solid tumors. Asian Pac J Cancer Prev. 15:8539–8548. 2014. View Article : Google Scholar : PubMed/NCBI
34	Pearson G, Robinson F, Gibson Beers T, Xu BE, Karandikar M, Berman K and Cobb MH: Mitogen-activated protein (MAP) kinase pathways: Regulation and physiological functions. Endocr Rev. 22:153–183. 2001. View Article : Google Scholar : PubMed/NCBI
35	Perugini RA, McDade TP, Vittimeberga FJ Jr and Callery MP: Pancreatic cancer cell proliferation is phosphatidylinositol 3-kinase dependent. J Surg Res. 90:39–44. 2000. View Article : Google Scholar : PubMed/NCBI
36	Tournier C: The 2 faces of JNK signaling in cancer. Genes Cancer. 4:397–400. 2013. View Article : Google Scholar : PubMed/NCBI
37	Takahashi R, Hirata Y, Sakitani K, Nakata W, Kinoshita H, Hayakawa Y, Nakagawa H, Sakamoto K, Hikiba Y, Ijichi H, et al: Theraputic effect of c-Jun N-terminal kinase inhibition on pancreatic cancer. Cancer Sci. 104:337–344. 2013. View Article : Google Scholar : PubMed/NCBI
38	Jia G, Kong R, Ma ZB, Han B, Wang YW, Pan SH, Li YH and Sun B: The activation of c-Jun NH₂-terminal kinase is required for dihydroartemisinin-induced autophagy in pancreatic cancer cells. J Exp Clin Cancer Res. 33:82014. View Article : Google Scholar : PubMed/NCBI
39	Li J, Liang X and Yang X: Ursolic acid inhibits growth and induces apoptosis in gemcitabine-resistant human pancreatic cancer via the JNK and PI3K/Akt/NF-kB pathways. Oncol Rep. 28:501–510. 2012.PubMed/NCBI