Britanin Exhibits Potential Inhibitory Activity on Human Prostate Cancer Cell Lines Through PI3K/Akt/NF-κB Signaling Pathways

Abstract

Britanin, a natural pseudoguaiacane sesquiterpene lactone, has significant antioxidant and anti-inflammatory activity, but little is known about its tumor inhibitory activity and the underlying mechanism. Here, we demonstrated in vitro and in vivo that britanin inhibited the growth of human prostate cancer cell lines (PC-3, PC-3-LUC, and DU-145). Through in vitro study, the results showed that britanin significantly decreased cell proliferation, migration, and motility. The moderate toxicity of britanin was determined with an acute toxicity study. A luciferase-labeled animal tumor xenograft model and bioluminescence imaging were applied, combining with biological validation for assessing the tumor progression. In vivo results demonstrated that britanin inhibited the growth of PC-3-LUC. The interleukin-2 level in mice was upregulated by britanin, which indicated that britanin induced antitumor immune activation. In addition, britanin downregulated the expression of nuclear factor (NF)-κB p105/p50, pp65, IκBα, pIκBα, phosphoinositide 3-kinase, pPI3k, Akt (protein kinase B, PKB), and pAkt proteins and upregulated expression of Bax. We discovered that britanin inhibits the growth of prostate cancer cells both in vitro and in vivo by regulating PI3K/Akt/NF-κB-related proteins and activating immunity. These findings shed light on the development of britanin as a promising agent for prostate cancer therapy.

Key words

britanin - prostate cancer - bioluminescence imaging - PI3K/Akt/NF-κB

Publication History

Received: 15 October 2019

Accepted after revision: 01 July 2020

Publication Date:
11 August 2020 (online)

• References

• 1 National Cancer Institute. SEER Cancer Statistics Fact sheets: prostate cancer. Online. Available at (Accessed February 22, 2020): https://seer.cancer.gov/statfacts/html/prost.html
• 2 Culp MB, Soerjomataram I, Efstathiou JA, Bray F, Jemal A. Recent global patterns in prostate cancer incidence and mortality rates. Eur Urol 2020; 77: 38-52
• 3 Litwin MS, Tan HJ. The diagnosis and treatment of prostate cancer: a review. JAMA 2017; 317: 2532-2542
• 4 Seca AML, Pinto DCGA. Plant secondary metabolites as anticancer agents: successes in clinical trials and therapeutic application. Int J Mol Sci 2018; 19: 263
• 5 Ghantous A, Gali-Muhtasib H, Vuorela H, Saliba NA, Darwiche N. What made sesquiterpene lactones reach cancer clinical trials?. Drug Discov Today 2010; 15: 668-678
• 6 Nie LY, Qin JJ, Huang Y, Yan L, Liu YB, Pan YX, Jin HZ, Zhang WD. Sesquiterpenoids from Inula lineariifolia inhibit nitric oxide production. J Nat Prod 2010; 73: 1117-1120
• 7 Kim SG, Lee E, Park NY, Park HH, Jeong KT, Kim KJ, Lee YJ, Jin MH, Lee E. Britanin attenuates ovalbumin-induced airway inflammation in a murine asthma model. Arch Pharm Res 2016; 39: 1006-1012
• 8 Wu GZ, Zhu LL, Yuan X, Chen H, Xiong R, Zhang SD, Cheng H, Shen YH, An HZ, Li TJ, Li HL, Zhang WD. Britanin ameliorates cerebral ischemia-reperfusion injury by inducing the Nrf2 protective pathway. Antioxid Redox Sign 2017; 27: 754-768
• 9 Klochkov SG, Pukhov SA, Afanasʼeva SV, Anikina LV, Ermatova AB. Amination products of Inula britannica lactones and their antitumor activity. Chem Nat Compd 2015; 51: 435-443
• 10 Lu Y, Li X, Park YN, Kwon O, Piao D, Chang YC, Kim CH, Lee E, Son JK, Chang HW. Britanin suppresses IgE/Ag-induced mast cell activation by inhibiting the syk pathway. Biomol Ther 2014; 22: 193-199
• 11 Park HH, Kim MJ, Li Y, Park YN, Lee J, Lee YJ, Kim SG, Park HJ, Son JK, Chang HW, Lee E. Britanin suppresses LPS-induced nitric oxide, PGE(2) and cytokine production via NF-kappa B and MAPK inactivation in RAW 264.7 cells. Int Immunopharmacol 2013; 15: 296-302
• 12 Beekman AC, Woerdenbag HJ, van Uden W, Pras N, Konings AWT, Wikstrom HV, Schmidt TJ. Structure-cytotoxicity relationships of some helenanolide-type sesquiterpene lactones. J Nat Prod 1997; 60: 252-257
• 13 Siedle B, Gustavsson L, Johansson S, Murillo R, Castro V, Bohlin L, Merfort I. The effect of sesquiterpene lactones on the release of human neutrophil elastase. Biochem Pharmacol 2003; 65: 897-903
• 14 Scotti MT, Fernandes MB, Ferreira MJP, Emerenciano VP. Quantitative structure-activity relationship of sesquiterpene lactones with cytotoxic activity. Bioorgan Med Chem 2007; 15: 2927-2934
• 15 Hall IH, Lee KH, Mar EC, Starnes CO, Waddell TG. Antitumor agents. 21. A proposed mechanism for inhibition of cancer growth by tenulin and helenalin and related cyclopentenones. J Med Chem 1977; 20: 333-337
• 16 Johnson JI, Decker S, Zaharevitz D, Rubinstein LV, Venditti J, Schepartz S, Kalyandrug S, Christian M, Arbuck S, Hollingshead M, Sausville EA. Relationships between drug activity in NCI preclinical in vitro and in vivo models and early clinical trials. Br J Cancer 2001; 84: 1424-1431
• 17 Willmann JK, van Bruggen N, Dinkelborg LM, Gambhir SS. Molecular imaging in drug development. Nat Rev Drug Discov 2008; 7: 591-607
• 18 Lin Y, Chen ZY, Yang F, Zhang JS, Wang YX, Liu JB, Liao JY, Liao YY, Zhou QL, Li BC, Liang HY. Application of molecular imaging technologies in antitumor drug development and therapy. Curr Pharm Design 2015; 21: 2136-2146
• 19 Fomchenko EI, Holland EC. Mouse models of brain tumors and their applications in preclinical trials. Clin Cancer Res 2006; 12: 5288-5297
• 20 Liang DS, Qin Y, Zhao WR, Zhai X, Guo ZW, Wang RX, Tong L, Lin LX, Chen H, Wong YC, Zhong ZH. S-allylmercaptocysteine effectively inhibits the proliferation of colorectal cancer cells under in vitro and in vivo conditions. Cancer Lett 2011; 310: 69-76
• 21 Xin J, Zhan YH, Liu MH, Hu H, Xia LM, Nie YZ, Wu KC, Liang JM, Tian J. ApoG2 induces ER stress-dependent apoptosis in gastric cancer cells in vitro and its real-time evaluation by bioluminescence imaging in vivo . Cancer Lett 2013; 336: 260-269
• 22 Wang Y, Zhang BL, Liu W, Dai YP, Shi YR, Zeng Q, Wang F. Noninvasive bioluminescence imaging of the dynamics of sanguinarine induced apoptosis via activation of reactive oxygen species. Oncotarget 2016; 7: 22355-22367
• 23 Amorim MHR, da Costa RMG, Lopes C, Bastos MMSM. Sesquiterpene lactones: adverse health effects and toxicity mechanisms. Crit Rev Toxicol 2013; 43: 559-579
• 24 Lissoni P. Therapy implications of the role of interleukin-2 in cancer. Expert Rev Clin Immu 2017; 13: 491-498
• 25 Kaarbo M, Mikkelsen OL, Malerod L, Qu S, Lobert VH, Akgul G, Halvorsen T, Maelandsmo GM, Saatcioglu F. PI3K-AKT-mTOR pathway is dominant over androgen receptor signaling in prostate cancer cells. Cell Oncol 2010; 32: 11-27
• 26 Chadwick M, Trewin H, Gawthrop F, Wagstaff C. Sesquiterpenoids Lactones: benefits to plants and people. Int J Mol Sci 2013; 14: 12780-12805
• 27 Formisano C, Sanna C, Ballero M, Chianese G, Sirignano C, Rigano D, Millan E, Munoz E, Taglialatela-Scafati O. Anti-inflammatory sesquiterpene lactones from Onopordum illyricum L. (Asteraceae), an Italian medicinal plant. Fitoterapia 2017; 116: 61-65
• 28 Salminen A, Lehtonen M, Suuronen T, Kaarniranta K, Huuskonen J. Terpenoids: natural inhibitors of NF-kappa B signaling with anti-inflammatory and anticancer potential. Cell Mol Life Sci 2008; 65: 2979-2999
• 29 Morais C, Gobe G, Johnson DW, Healy H. The emerging role of nuclear factor kappa B in renal cell carcinoma. Int J Biochem Cell B 2011; 43: 1537-1549
• 30 Glauert HP, Tharappel JC, Banerjee S, Chan NLS, Kania-Korwel I, Lehmler HJ, Lee EY, Robertson LW, Spear BT. Inhibition of the promotion of hepatocarcinogenesis by 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB-153) by the deletion of the p50 subunit of NF-kappa B in mice. Toxicol Appl Pharm 2008; 232: 302-308
• 31 Zhang L, Tao L, Ruan JS, Li WD, Wu Y, Yan LG, Zhang F, Fan FT, Zheng SZ, Wang AY, Lu Y. Xanthatin induces G2/M cell cycle arrest and apoptosis in human gastric carcinoma MKN-45 cells. Planta Med 2012; 78: 890-895
• 32 Bitting RL, Armstrong AJ. Targeting the PI3K/Akt/mTOR pathway in castration-resistant prostate cancer. Endocr-Relat Cancer 2013; 20: R83-R99
• 33 Zheng HC. The molecular mechanisms of chemoresistance in cancers. Oncotarget 2017; 8: 59950-59964
• 34 Choi ES, Kim JS, Kwon KH, Kim HS, Cho NP, Cho SD. Methanol extract of Sanguisorba officinalis L. with cytotoxic activity against PC3 human prostate cancer cells. Mol Med Rep 2012; 6: 670-674
• 35 Lowe SL, Rubinchik S, Honda T, McDonnell TJ, Dong JY, Norris JS. Prostate-specific expression of Bax delivered by an adenoviral vector induces apoptosis in LNCaP prostate cancer cells. Gene Ther 2001; 8: 1363-1371
• 36 Strobel T, Kraeft SK, Chen LB, Cannistra SA. BAX expression is associated with enhanced intracellular accumulation of paclitaxel: a novel role for BAX during chemotherapy-induced cell death. Cancer Res 1998; 58: 4776-4781
• 37 Chai WS, Zhu XM, Li SH, Fan JX, Chen BY. Role of Bcl-2 family members in caspase-3/9-dependent apoptosis during Pseudomonas aeruginosa infection in U937 cells. Apoptosis 2008; 13: 833-843
• 38 Arenas-Ramirez N, Woytschak J, Boyman O. Interleukin-2: biology, design and application. Trends Immunol 2015; 36: 763-777
• 39 Gomez J, Garcia-Domingo D, Martinez AC, Rebollo A. Role of NF-kappaB in the control of apoptotic and proliferative responses in IL-2-responsive T cells. Front Biosci 1997; 2: d49-d60
• 40 Lehr HA, Mankoff DA, Corwin D, Santeusanio G, Gown AM. Application of photoshop-based image analysis to quantification of hormone receptor expression in breast cancer. J Histochem Cytochem 1997; 45: 1559-1565
• 41 Xu JH, Liu DY, Niu HL, Zhu GF, Xu YW, Ye DL, Li J, Zhang QL. Resveratrol reverses Doxorubicin resistance by inhibiting epithelial-mesenchymal transition (EMT) through modulating PTEN/Akt signaling pathway in gastric cancer. J Exp Clin Canc Res 2017; 36: 19
• 42 Li K, Liu H, Gao W, Chen M, Zeng Y, Liu JJ, Xu L, Wu DC. Mulberry-like dual-drug complicated nanocarriers assembled with apogossypolone amphiphilic starch micelles and doxorubicin hyaluronic acid nanoparticles for tumor combination and targeted therapy. Biomaterials 2015; 39: 131-144
• 43 Alleva E, Santucci D. Guide for the care and use of laboratory animals. Ethology 1997; 103: 1072-1073
• 44 Li HR, Li K, Dai YP, Xu XY, Cao X, Zeng Q, He HYL, Pang LJ, Liang JM, Chen XL, Zhan YH. In vivo near infrared fluorescence imaging and dynamic quantification of pancreatic metastatic tumors using folic acid conjugated biodegradable mesoporous silica nanoparticles. Nanomed-Nanotechnol 2018; 14: 1867-1877