Frankincense essential oil prepared from hydrodistillation of Boswellia sacra gum resins induces human pancreatic cancer cell death in cultures and in a xenograft murine model

BMC Complementary and Alternative Medicine

Hsueh-Kung Lin²  Jingzhe Zhang⁷  Gary Young⁵  Cole Woolley⁵  Russell G Postier⁴  Kar-Ming Fung³  Amy Cao¹  Qing Yang²  Mahmoud M Suhail⁶  Xiao Ni⁷ 

[1] Department of Biological Sciences, University of Southern California, Los Angeles, CA, 90089, USA;Department of Urology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA;Oklahoma City Veterans Medical Center, Oklahoma City, OK, 73104, USA;Department of Surgery, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA;Young Living Essential Oils, Lehi, UT, 84043, USA;Al Afia Medical Complex, Salalah, Sultanate of Oman;Department of General Surgery, Long Hua Hospital Shanghai University of Traditional Chinese Medicine, Shanghai, China

关键词: Pancreatic cancer;    Hydrodistillation;    Frankincense;    Essential oil;    Boswellia sacra, Boswellic acid;    Apoptosis;   

Others  :  1231372
DOI  :  10.1186/1472-6882-12-253

 received in 2012-04-24, accepted in 2012-12-11,  发布年份 2012

【 摘 要 】

Background

Regardless of the availability of therapeutic options, the overall 5-year survival for patients diagnosed with pancreatic cancer remains less than 5%. Gum resins from Boswellia species, also known as frankincense, have been used as a major ingredient in Ayurvedic and Chinese medicine to treat a variety of health-related conditions. Both frankincense chemical extracts and essential oil prepared from Boswellia species gum resins exhibit anti-neoplastic activity, and have been investigated as potential anti-cancer agents. The goals of this study are to identify optimal condition for preparing frankincense essential oil that possesses potent anti-tumor activity, and to evaluate the activity in both cultured human pancreatic cancer cells and a xenograft mouse cancer model.

Methods

Boswellia sacra gum resins were hydrodistilled at 78°C; and essential oil distillate fractions were collected at different durations (Fraction I at 0–2 h, Fraction II at 8–10 h, and Fraction III at 11–12 h). Hydrodistillation of the second half of gum resins was performed at 100°C; and distillate was collected at 11–12 h (Fraction IV). Chemical compositions were identified by gas chromatography–mass spectrometry (GC-MS); and total boswellic acids contents were quantified by high-performance liquid chromatography (HPLC). Frankincense essential oil-modulated pancreatic tumor cell viability and cytotoxicity were determined by colorimetric assays. Levels of apoptotic markers, signaling molecules, and cell cycle regulators expression were characterized by Western blot analysis. A heterotopic (subcutaneous) human pancreatic cancer xenograft nude mouse model was used to evaluate anti-tumor capability of Fraction IV frankincense essential oil in vivo. Frankincense essential oil-induced tumor cytostatic and cytotoxic activities in animals were assessed by immunohistochemistry.

Results

Longer duration and higher temperature hydrodistillation produced more abundant high molecular weight compounds, including boswellic acids, in frankincense essential oil fraactions. Human pancreatic cancer cells were sensitive to Fractions III and IV (containing higher molecular weight compounds) treatment with suppressed cell viability and increased cell death. Essential oil activated the caspase-dependent apoptotic pathway, induced a rapid and transient activation of Akt and Erk1/2, and suppressed levels of cyclin D1 cdk4 expression in cultured pancreatic cancer cells. In addition, Boswellia sacra essential oil Fraction IV exhibited anti-proliferative and pro-apoptotic activities against pancreatic tumors in the heterotopic xenograft mouse model.

Conclusion

All fractions of frankincense essential oil from Boswellia sacra are capable of suppressing viability and inducing apoptosis of a panel of human pancreatic cancer cell lines. Potency of essential oil-suppressed tumor cell viability may be associated with the greater abundance of high molecular weight compounds in Fractions III and IV. Although chemical component(s) responsible for tumor cell cytotoxicity remains undefined, crude essential oil prepared from hydrodistillation of Boswellia sacra gum resins might be a useful alternative therapeutic agent for treating patients with pancreatic adenocarcinoma, an aggressive cancer with poor prognosis.

【 授权许可】

   
2012 Ni et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Siegel R, Naishadham D, Jemal A: Cancer statistics, 2012. CA Cancer J Clin 2012, 62(1):10-29.
• [2]Hariharan D, Saied A, Kocher HM: Analysis of mortality rates for pancreatic cancer across the world. HPB (Oxford) 2008, 10(1):58-62.
• [3]Nakai Y, Isayama H, Sasaki T, Sasahira N, Tsujino T, Toda N, Kogure H, Matsubara S, Ito Y, Togawa O, et al.: A multicentre randomised phase II trial of gemcitabine alone vs gemcitabine and S-1 combination therapy in advanced pancreatic cancer: GEMSAP study. Br J Cancer 2012, 106(12):1934-1939.
• [4]Neoptolemos JP, Stocken DD, Bassi C, Ghaneh P, Cunningham D, Goldstein D, Padbury R, Moore MJ, Gallinger S, Mariette C, et al.: Adjuvant chemotherapy with fluorouracil plus folinic acid vs gemcitabine following pancreatic cancer resection: a randomized controlled trial. JAMA 2010, 304(10):1073-1081.
• [5]Gebbia V, Maiello E, Giuliani F, Borsellino N, Arcara C, Colucci G: Irinotecan plus bolus/infusional 5-Fluorouracil and leucovorin in patients with pretreated advanced pancreatic carcinoma: a multicenter experience of the Gruppo Oncologico Italia Meridionale. Am J Clin Oncol 2010, 33(5):461-464.
• [6]Jeon EK, Won HS, Ko YH, Lee IS, Hong TH, You YK, Lee MA: Comparison of the efficacy and the toxicity between gemcitabine with capecitabine (GC) and gemcitabine with erlotinib (GE) in unresectable pancreatic cancer. J Cancer Res Clin Oncol 2012, 138(10):1625-1630.
• [7]Winking M, Sarikaya S, Rahmanian A, Jodicke A, Boker DK: Boswellic acids inhibit glioma growth: a new treatment option? J Neurooncol 2000, 46(2):97-103.
• [8]Huang MT, Badmaev V, Ding Y, Liu Y, Xie JG, Ho CT: Anti-tumor and anti-carcinogenic activities of triterpenoid, b-boswellic acid. Biofactors 2000, 13(1–4):225-230.
• [9]Kirste S, Treier M, Wehrle SJ, Becker G, Abdel-Tawab M, Gerbeth K, Hug MJ, Lubrich B, Grosu AL, Momm F: Boswellia serrata acts on cerebral edema in patients irradiated for brain tumors: a prospective, randomized, placebo-controlled, double-blind pilot trial. Cancer 2011, 117(16):3788-3795.
• [10]Frank MB, Yang Q, Osban J, Azzarello JT, Saban MR, Saban R, Ashley RA, Welter JC, Fung KM, Lin HK: Frankincense oil derived from Boswellia carteri induces tumor cell specific cytotoxicity. BMC Complement Altern Med 2009, 9:6. BioMed Central Full Text
• [11]Suhail MM, Wu W, Cao A, Mondalek FG, Fung KM, Shih PT, Fang YT, Woolley C, Young G, Lin HK: Boswellia sacra essential oil induces tumor cell-specific apoptosis and suppresses tumor aggressiveness in cultured human breast cancer cells. BMC Complement Altern Med 2011, 11:129. BioMed Central Full Text
• [12]Yunis AA, Arimura GK, Russin DJ: Human pancreatic carcinoma (MIA PaCa-2) in continuous culture: sensitivity to asparaginase. Int J Cancer 1977, 19(1):128-135.
• [13]Frazier ML, Fernandez E, de Llorens R, Brown NM, Pathak S, Cleary KR, Abbruzzese JL, Berry K, Olive M, Le Maistre A, et al.: Pancreatic adenocarcinoma cell line, MDAPanc-28, with features of both acinar and ductal cells. Int J Pancreatol 1996, 19(1):31-38.
• [14]Tan MH, Nowak NJ, Loor R, Ochi H, Sandberg AA, Lopez C, Pickren JW, Berjian R, Douglass HO Jr, Chu TM: Characterization of a new primary human pancreatic tumor line. Cancer Invest 1986, 4(1):15-23.
• [15]Rosewicz S, Stier U, Brembeck F, Kaiser A, Papadimitriou CA, Berdel WE, Wiedenmann B, Riecken EO: Retinoids: effects on growth, differentiation, and nuclear receptor expression in human pancreatic carcinoma cell lines. Gastroenterology 1995, 109(5):1646-1660.
• [16]Mondalek FG, Lawrence BJ, Kropp BP, Grady BP, Fung KM, Madihally SV, Lin HK: The incorporation of poly(lactic-co-glycolic) acid nanoparticles into porcine small intestinal submucosa biomaterials. Biomaterials 2008, 29(9):1159-1166.
• [17]Boehm T, Folkman J, Browder T, O’Reilly MS: Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 1997, 390(6658):404-407.
• [18]Amoh Y, Li L, Yang M, Jiang P, Moossa AR, Katsuoka K, Hoffman RM: Hair follicle-derived blood vessels vascularize tumors in skin and are inhibited by Doxorubicin. Cancer Res 2005, 65(6):2337-2343.
• [19]Fung KM, Samara ENS, Wong C, Metwalli A, Krlin R, Bane B, Liu CZ, Yang JT, Pitha JT, Culkin DJ, et al.: Increased expression of type 2 3a-hydroxysteroid dehydrogenase/type 5 17b-hydroxysteroid dehydrogenase (AKR1C3) and its relationship with androgen receptor in prostate carcinoma. Endocr Relat Cancer 2006, 13(1):169-180.
• [20]Yang Q, Titus M, Fung KM, Lin HK: 5α-Androstane-3α,17β-diol supports human prostate cancer cell survival and proliferation through androgen receptor-independent signaling pathways: Implication of androgen-independent prostate cancer progression. J Cell Biochem 2008, 104(5):1612-1624.
• [21]Satoh MS, Lindahl T: Role of poly(ADP-ribose) formation in DNA repair. Nature 1992, 356(6367):356-358.
• [22]Oliver FJ, de la Rubia G, Rolli V, Ruiz-Ruiz MC, de Murcia G, Murcia JM: Importance of poly(ADP-ribose) polymerase and its cleavage in apoptosis. Lesson from an uncleavable mutant. J Biol Chem 1998, 273(50):33533-33539.
• [23]Tapia C, Kutzner H, Mentzel T, Savic S, Baumhoer D, Glatz K: Two mitosis-specific antibodies, MPM-2 and phospho-histone H3 (Ser28), allow rapid and precise determination of mitotic activity. Am J Surg Pathol 2006, 30(1):83-89.
• [24]Gavrieli Y, Sherman Y, Ben-Sasson SA: Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 1992, 119(3):493-501.
• [25]Hostanska K, Daum G, Saller R: Cytostatic and apoptosis-inducing activity of boswellic acids toward malignant cell lines in vitro. Anticancer Res 2002, 22(5):2853-2862.
• [26]Akihisa T, Tabata K, Banno N, Tokuda H, Nishimura R, Nakamura Y, Kimura Y, Yasukawa K, Suzuki T: Cancer chemopreventive effects and cytotoxic activities of the triterpene acids from the resin of Boswellia carteri. Biol Pharm Bull 2006, 29(9):1976-1979.
• [27]Shao Y, Ho CT, Chin CK, Badmaev V, Ma W, Huang MT: Inhibitory activity of boswellic acids from Boswellia serrata against human leukemia HL-60 cells in culture. Planta Med 1998, 64(4):328-331.
• [28]Park YS, Lee JH, Bondar J, Harwalkar JA, Safayhi H, Golubic M: Cytotoxic action of acetyl-11-keto-b-boswellic acid (AKBA) on meningioma cells. Planta Med 2002, 68(5):397-401.
• [29]Xia L, Chen D, Han R, Fang Q, Waxman S, Jing Y: Boswellic acid acetate induces apoptosis through caspase-mediated pathways in myeloid leukemia cells. Mol Cancer Ther 2005, 4(3):381-388.
• [30]Liu JJ, Nilsson A, Oredsson S, Badmaev V, Duan RD: Keto- and acetyl-keto-boswellic acids inhibit proliferation and induce apoptosis in Hep G2 cells via a caspase-8 dependent pathway. Int J Mol Med 2002, 10(4):501-505.
• [31]Zhao W, Entschladen F, Liu H, Niggemann B, Fang Q, Zaenker KS, Han R: Boswellic acid acetate induces differentiation and apoptosis in highly metastatic melanoma and fibrosarcoma cells. Cancer Detect Prev 2003, 27(1):67-75.
• [32]Liu JJ, Nilsson A, Oredsson S, Badmaev V, Zhao WZ, Duan RD: Boswellic acids trigger apoptosis via a pathway dependent on caspase-8 activation but independent on Fas/Fas ligand interaction in colon cancer HT-29 cells. Carcinogenesis 2002, 23(12):2087-2093.
• [33]Liu JJ, Huang B, Hooi SC: Acetyl-keto-b-boswellic acid inhibits cellular proliferation through a p21-dependent pathway in colon cancer cells. Br J Pharmacol 2006, 148(8):1099-1107.
• [34]Pang X, Yi Z, Zhang X, Sung B, Qu W, Lian X, Aggarwal BB, Liu M: Acetyl-11-keto-b-boswellic acid inhibits prostate tumor growth by suppressing vascular endothelial growth factor receptor 2-mediated angiogenesis. Cancer Res 2009, 69(14):5893-5900.
• [35]Lu M, Xia L, Hua H, Jing Y: Acetyl-keto-b-boswellic acid induces apoptosis through a death receptor 5-mediated pathway in prostate cancer cells. Cancer Res 2008, 68(4):1180-1186.
• [36]Park B, Prasad S, Yadav V, Sung B, Aggarwal BB: Boswellic acid suppresses growth and metastasis of human pancreatic tumors in an orthotopic nude mouse model through modulation of multiple targets. PLoS One 2011, 6(10):e26943.
• [37]Park B, Sung B, Yadav VR, Cho SG, Liu M, Aggarwal BB: Acetyl-11-keto-b-boswellic acid suppresses invasion of pancreatic cancer cells through the downregulation of CXCR4 chemokine receptor expression. Int J Cancer 2011, 129(1):23-33.
• [38]Estrada AC, Syrovets T, Pitterle K, Lunov O, Buchele B, Schimana-Pfeifer J, Schmidt T, Morad SA, Simmet T: Tirucallic acids are novel pleckstrin homology domain-dependent Akt inhibitors inducing apoptosis in prostate cancer cells. Mol Pharmacol 2010, 77(3):378-387.
• [39]Syrovets T, Gschwend JE, Buchele B, Laumonnier Y, Zugmaier W, Genze F, Simmet T: Inhibition of IkB kinase activity by acetyl-boswellic acids promotes apoptosis in androgen-independent PC-3 prostate cancer cells in vitro and in vivo. J Biol Chem 2005, 280(7):6170-6180.
• [40]Glaser T, Winter S, Groscurth P, Safayhi H, Sailer ER, Ammon HP, Schabet M, Weller M: Boswellic acids and malignant glioma: induction of apoptosis but no modulation of drug sensitivity. Br J Cancer 1999, 80(5–6):756-765.
• [41]Hoernlein RF, Orlikowsky T, Zehrer C, Niethammer D, Sailer ER, Simmet T, Dannecker GE, Ammon HP: Acetyl-11-keto-b-boswellic acid induces apoptosis in HL-60 and CCRF-CEM cells and inhibits topoisomerase I. J Pharmacol Exp Ther 1999, 288(2):613-619.
• [42]Quelle DE, Ashmun RA, Shurtleff SA, Kato JY, Bar-Sagi D, Roussel MF, Sherr CJ: Overexpression of mouse D-type cyclins accelerates G1 phase in rodent fibroblasts. Genes Dev 1993, 7(8):1559-1571.
• [43]Fahraeus R, Paramio JM, Ball KL, Lain S, Lane DP: Inhibition of pRb phosphorylation and cell-cycle progression by a 20-residue peptide derived from p16CDKN2/INK4A. Curr Biol 1996, 6(1):84-91.
• [44]Dupont J, Karas M, LeRoith D: The cyclin-dependent kinase inhibitor p21CIP/WAF is a positive regulator of insulin-like growth factor I-induced cell proliferation in MCF-7 human breast cancer cells. J Biol Chem 2003, 278(39):37256-37264.
• [45]Liu JJ, Duan RD: LY294002 enhances boswellic acid-induced apoptosis in colon cancer cells. Anticancer Res 2009, 29(8):2987-2991.
• [46]Altmann A, Fischer L, Schubert-Zsilavecz M, Steinhilber D, Werz O: Boswellic acids activate p42(MAPK) and p38 MAPK and stimulate Ca(2+) mobilization. Biochem Biophys Res Commun 2002, 290(1):185-190.
• [47]Poeckel D, Tausch L, Altmann A, Feisst C, Klinkhardt U, Graff J, Harder S, Werz O: Induction of central signalling pathways and select functional effects in human platelets by b-boswellic acid. Br J Pharmacol 2005, 146(4):514-524.
• [48]Wang L, Wang P, Liu Y, Xue Y: Regulation of cellular growth, apoptosis, and Akt activity in human U251 glioma cells by a combination of cisplatin with CRM197. Anticancer Drugs 2012, 23(1):81-89.
• [49]Lee YJ, Jeong HY, Kim YB, Won SY, Shim JH, Cho MK, Nam HS, Lee SH: Reactive oxygen species and PI3K/Akt signaling play key roles in the induction of Nrf2-driven heme oxygenase-1 expression in sulforaphane-treated human mesothelioma MSTO-211H cells. Food Chem Toxicol 2012, 50(2):116-123.
• [50]Kumar A, Shah BA, Singh S, Hamid A, Singh SK, Sethi VK, Saxena AK, Singh J, Taneja SC: Acyl derivatives of boswellic acids as inhibitors of NF-kB and STATs. Bioorg Med Chem Lett 2012, 22(1):431-435.
• [51]Holtmeier W, Zeuzem S, Preiss J, Kruis W, Bohm S, Maaser C, Raedler A, Schmidt C, Schnitker J, Schwarz J, et al.: Randomized, placebo-controlled, double-blind trial of Boswellia serrata in maintaining remission of Crohn’s disease: good safety profile but lack of efficacy. Inflamm Bowel Dis 2011, 17(2):573-582.
• [52]Krishnaraju AV, Sundararaju D, Vamsikrishna U, Suryachandra R, Machiraju G, Sengupta K, Trimurtulu G: Safety and toxicological evaluation of Aflapin: a novel Boswellia-derived anti-inflammatory product. Toxicol Mech Methods 2010, 20(9):556-563.
• [53]Sengupta K, Alluri KV, Satish AR, Mishra S, Golakoti T, Sarma KV, Dey D, Raychaudhuri SP: A double blind, randomized, placebo controlled study of the efficacy and safety of 5-Loxin for treatment of osteoarthritis of the knee. Arthritis Res Ther 2008, 10(4):R85. BioMed Central Full Text
• [54]Lalithakumari K, Krishnaraju AV, Sengupta K, Subbaraju GV, Chatterjee A: Safety and Toxicological Evaluation of a Novel, Standardized 3-O-Acetyl-11-keto-beta-Boswellic Acid (AKBA)-Enriched Boswellia serrata Extract (5-Loxin(R)). Toxicol Mech Methods 2006, 16(4):199-226.
• [55]Kimmatkar N, Thawani V, Hingorani L, Khiyani R: Efficacy and tolerability of Boswellia serrata extract in treatment of osteoarthritis of knee–a randomized double blind placebo controlled trial. Phytomedicine 2003, 10(1):3-7.