Ethyl acetate extract from Inula helenium L. inhibits the proliferation of pancreatic cancer cells by regulating the STAT3/AKT pathway

Zhang,Bo; Zeng,Jianmei; Yan,Youyou; Yang,Bo; Huang,Mincong; Wang,Linling; Zhang,Qi; Lin,Nengming

doi:10.3892/mmr.2018.8534

Ethyl acetate extract from Inula helenium L. inhibits the proliferation of pancreatic cancer cells by regulating the STAT3/AKT pathway

Authors:
- Bo Zhang
- Jianmei Zeng
- Youyou Yan
- Bo Yang
- Mincong Huang
- Linling Wang
- Qi Zhang
- Nengming Lin
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Affiliations: Translational Medicine Research Center, Hangzhou First People's Hospital, Nanjing Medical University, Hangzhou, Zhejiang 310006, P.R. China, College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, P.R. China
Published online on: February 1, 2018 https://doi.org/10.3892/mmr.2018.8534
Pages: 5440-5448

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Abstract

Sesquiterpene lactones are bioactive compounds that have been identified as responsible for the anticancer activity of the medicinal herb, Inula helenium L. (IHL). However, the mechanisms of action involved in the anti‑pancreatic cancer activity of IHL have yet to be elucidated. The present study used an optimized extraction strategy to obtain sesquiterpene lactones from IHL (the resulting product termed ethyl acetate extract of IHL; EEIHL), and examined the potential mechanisms involved in the anti‑pancreatic cancer activity of EEIHL. Ethanol and ethyl acetate were used to extract sesquiterpene lactones from IHL to give the final product EEIHL. Cell Counting Kit‑8, colony formation and Annexin V/propidium iodide assays were used to detect the anti‑proliferative activity of EEIHL. Cell migration was determined with a wound healing assay. mRNA and protein expression levels were analyzed by reverse transcription‑quantitative polymerase chain reaction and western blot analyses, respectively. It was identified that low concentrations of EEIHL caused CFPAC‑1 cell cycle arrest in the G0/G1 phase, whereas high concentrations of EEIHL induced mitochondria‑dependent apoptosis. In addition, EEIHL could inhibit the phosphorylation of the signal transducer and activator of transcription (STAT)3/AKT pathway, potentially resulting in impeded cell mobility. In conclusion, EEIHL could activate mitochondrial‑dependent apoptosis and inhibit cell migration through the STAT3/AKT pathway in CFPAC-1 cells.

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1	Rahib L, Smith BD, Aizenberg 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. Can Res. 74:2913–2921. 2014. View Article : Google Scholar
2	Bailey P, Chang DK, Nones K, Johns AL, Patch AM, Gingras MC, Miller DK, Christ AN, Bruxner TJ, Quinn MC, et al: Genomic analyses identify molecular subtypes of pancreatic cancer. Nature. 531:47–52. 2016. View Article : Google Scholar : PubMed/NCBI
3	Garrido-Laguna I and Hidalgo M: Pancreatic cancer: From state-of-the-art treatments to promising novel therapies. Nat Rev Clin Oncol. 12:319–334. 2015. View Article : Google Scholar : PubMed/NCBI
4	Vincent A, Herman J, Schulick R, Hruban RH and Goggins M: Pancreatic cancer. Lancet. 378:607–620. 2011. View Article : Google Scholar : PubMed/NCBI
5	Seca AM, Grigore A, Pinto DC and Silva AM: The genus Inula and their metabolites: From ethnopharmacological to medicinal uses. J Ethnopharmacol. 154:286–310. 2014. View Article : Google Scholar : PubMed/NCBI
6	Stojanović-Radić Z, Comić LJ, Radulović N, Blagojević P, Denić M, Miltojević A, Rajković J and Mihajilov-Krstev T: Antistaphylococcal activity of Inula helenium L. root essential oil: Eudesmane sesquiterpene lactones induce cell membrane damage. Eur J Clin Microbiol Infect Dis. 31:1015–1025. 2012. View Article : Google Scholar : PubMed/NCBI
7	Qiu J, Luo M, Wang J, Dong J, Li H, Leng B, Zhang Q, Dai X, Zhang Y, Niu X and Deng X: Isoalantolactone protects against Staphylococcus aureus pneumonia. FEMS Microbiol Lett. 324:147–155. 2011. View Article : Google Scholar : PubMed/NCBI
8	Ghantous A, Gali-Muhtasib H, Vuorela H, Saliba NA and Darwiche N: What made sesquiterpene lactones reach cancer clinical trials? Drug Discov Today. 15:668–678. 2010. View Article : Google Scholar : PubMed/NCBI
9	Jiang HL, Chen J, Jin XJ, Yang JL, Li Y, Yao XJ and Wu QX: Sesquiterpenoids, alantolactone analogues, and seco-guaiene from the roots of Inula helenium. Tetrahedron. 67:9193–9198. 2011. View Article : Google Scholar
10	Zhang S, Won YK, Ong CN and Shen HM: Anti-cancer potential of sesquiterpene lactones: Bioactivity and molecular mechanisms. Curr Med Chem. 5:239–249. 2005.
11	Wang J, Zhao YM, Tian YT, Yan CL and Guo CY: Ultrasound-assisted extraction of total phenolic compounds from Inula helenium. ScientificWorldJournal. 2013:1575272013.PubMed/NCBI
12	Park EJ, Kim YM, Park SW, Kim HJ, Lee JH, Lee DU and Chang KC: Induction of HO-1 through p38 MAPK/Nrf2 signaling pathway by ethanol extract of Inula helenium L. reduces inflammation in LPS-activated RAW 264.7 cells and CLP-induced septic mice. Food Chem Toxicol. 55:386–395. 2013. View Article : Google Scholar : PubMed/NCBI
13	Guo CY, Wang J, Hou Y, Zhao YM, Shen LX and Zhang DS: Orthogonal test design for optimizing the extraction of total flavonoids from Inula helenium. Pharmacogn Mag. 9:192–195. 2013. View Article : Google Scholar : PubMed/NCBI
14	Ahmed AF, Al-Qahtani JH, Al-Yousef HM, Al-Said MS, Ashour AE, Al-Sohaibani M and Rafatullah S: Proanthocyanidin-rich date seed extract protects against chemically induced hepatorenal toxicity. J Med Food. 18:280–289. 2015. View Article : Google Scholar : PubMed/NCBI
15	Du J, Shi HR, Ren F, Wang JL, Wu QH, Li X and Zhang RT: Inhibition of the IGF signaling pathway reverses cisplatin resistance in ovarian cancer cells. BMC Cancer. 17:8512017. View Article : Google Scholar : PubMed/NCBI
16	Ding YL, Wang YZ, Zhang J, Zhang QZ, Zhang JY and Jin H: Application of the vanillin sulfuric acid colorimetry-ultraviolet spectrometry on quality evaluation of Panax notoginseng. Guang Pu Xue Yu Guang Pu Fen Xi. 33:471–475. 2013.(In Chines). PubMed/NCBI
17	Konishi T, Shimada Y, Nagao T, Okabe H and Konoshima T: Antiproliferative sesquiterpene lactones from the roots of Inula helenium. Biol Pharm Bull. 25:1370–1372. 2002. View Article : Google Scholar : PubMed/NCBI
18	Weng Z, Gao H, Hu J, Fan Y, Wang H and Li L: Isoalantolactone induces autophagic cell death in SKOV3 human ovarian carcinoma cells via upregulation of PEA-15. Oncol Rep. 35:833–840. 2016. View Article : Google Scholar : PubMed/NCBI
19	Di W, Khan M, Rasul A, Sun M, Sui Y, Zhong L, Yang L, Zhu Q, Feng L and Ma T: Isoalantolactone inhibits constitutive NF-κB activation and induces reactive oxygen species-mediated apoptosis in osteosarcoma U2OS cells through mitochondrial dysfunction. Oncol Rep. 32:1585–1593. 2014. View Article : Google Scholar : PubMed/NCBI
20	Zaima K, Wakana D, Demizu Y, Kumeta Y, Kamakura H, Maruyama T, Kurihara M and Goda Y: Isoheleproline: A new amino acid-sesquiterpene adduct from Inula helenium. J Nat Med. 68:432–435. 2014. View Article : Google Scholar : PubMed/NCBI
21	Chen CN, Huang HH, Wu CL, Lin CP, Hsu JT, Hsieh HP, Chuang SE and Lai GM: Isocostunolide, a sesquiterpene lactone, induces mitochondrial membrane depolarization and caspase-dependent apoptosis in human melanoma cells. Cancer Lett. 246:237–252. 2007. View Article : Google Scholar : PubMed/NCBI
22	Zhao YM, Wang J, Liu HB, Guo CY and Zhang WM: Microwave-assisted Extraction of Alantolactone and Isoalantolactone from Inula helenium. Indian J Pharm Sci. 77:116–120. 2015. View Article : Google Scholar : PubMed/NCBI
23	Cai H, Meng X, Li Y, Yang C and Liu Y: Growth inhibition effects of isoalantolactone on K562/A02 cells: Caspase-dependent apoptotic pathways, S phase arrest, and downregulation of Bcr/Abl. Phytother Res. 28:1679–1686. 2014. View Article : Google Scholar : PubMed/NCBI
24	Wang J, Cui L, Feng L, Zhang Z, Song J, Liu D and Jia X: Isoalantolactone inhibits the migration and invasion of human breast cancer MDA-MB-231 cells via suppression of the p38 MAPK/NF-κB signaling pathway. Oncol Rep. 36:1269–1276. 2016. View Article : Google Scholar : PubMed/NCBI
25	Khan M, Ding C, Rasul A, Yi F, Li T, Gao H, Gao R, Zhong L, Zhang K, Fang X and Ma T: Isoalantolactone induces reactive oxygen species mediated apoptosis in pancreatic carcinoma PANC-1 cells. Int J Biol Sci. 8:533–547. 2012. View Article : Google Scholar : PubMed/NCBI
26	Zhang M, Hou Y, Shen Y, Guo X, Shang D and Zhang D: Probucol reverses homocysteine induced inflammatory monocytes differentiation and oxidative stress. Eur J Pharmacol. 818:67–73. 2018. View Article : Google Scholar : PubMed/NCBI
27	Yao Y, Xia D, Bian Y, Sun Y, Zhu F, Pan B, Niu M, Zhao K, Wu Q, Qiao J, et al: Alantolactone induces G1 phase arrest and apoptosis of multiple myeloma cells and overcomes bortezomib resistance. Apoptosis. 20:1122–1133. 2015. View Article : Google Scholar : PubMed/NCBI
28	Chen YW, Hsiao PJ, Weng CC, Kuo KK, Kuo TL, Wu DC, Hung WC and Cheng KH: SMAD4 loss triggers the phenotypic changes of pancreatic ductal adenocarcinoma cells. BMC Cancer. 14:1812014. View Article : Google Scholar : PubMed/NCBI
29	Zhang X, Cao J, Pei Y, Zhang J and Wang Q: Smad4 inhibits cell migration via suppression of JNK activity in human pancreatic carcinoma PANC-1 cells. Oncol Lett. 11:3465–3470. 2016. View Article : Google Scholar : PubMed/NCBI
30	Yang G, Murashige DS, Humphrey SJ and James DE: A positive feedback loop between Akt and mTORC2 via SIN1 phosphorylation. Cell Rep. 12:937–943. 2015. View Article : Google Scholar : PubMed/NCBI
31	Sarbassov DD, Guertin DA, Ali SM and Sabatini DM: Phosphorylation and Regulation of Akt/PKB by the Rictor-mTOR Complex. Science. 307:1098–1101. 2005. View Article : Google Scholar : PubMed/NCBI
32	Guo Y, Zang Y, Lv L, Cai F, Qian T, Zhang G and Feng Q: IL-8 promotes proliferation and inhibition of apoptosis via STAT3/AKT/NF-κB pathway in prostate cancer. Mol Med Rep. 16:9035–9042. 2017. View Article : Google Scholar : PubMed/NCBI
33	Saitoh M, Endo K, Furuya S, Minami M, Fukasawa A, Imamura T and Miyazawa K: STAT3 integrates cooperative Ras and TGF-β signals that induce Snail expression. Oncogene. 35:1049–1057. 2016. View Article : Google Scholar : PubMed/NCBI
34	Yu M, Xue H, Wang Y, Shen Q, Jiang Q, Zhang X, Li K, Jia M, Jia J, Xu J and Tian Y: miR-345 inhibits tumor metastasis and EMT by targeting IRF1-mediated mTOR/STAT3/AKT pathway in hepatocellular carcinoma. Int J Oncol. 50:975–983. 2017. View Article : Google Scholar : PubMed/NCBI
35	Kim HN, Narayanan NK, Lasano S and Narayanan B: Modulation of PGE2-induced EP4 expression on snail signaling and the impact on epithelial-mesenchymal transition: Significance of EP4 antagonism. Anticancer Res. 31:4347–4357. 2011.PubMed/NCBI
36	Shi Y, Zuo D, Wang X, Han M and Wu Y: shRNA-mediated silencing of TARBP2 inhibits NCI-H1299 non-small cell lung cancer cell invasion and migration via the JNK/STAT3/AKT pathway. Mol Med Rep. 14:3725–3730. 2016. View Article : Google Scholar : PubMed/NCBI