Phytochemical Analysis and Anti-cancer Investigation of Boswellia Serrata Bioactive Constituents In Vitro

  • Ahmed, Hanaa H (Hormones Department, National Research Centre) ;
  • Abd-Rabou, Ahmed A (Hormones Department, National Research Centre) ;
  • Hassan, Amal Z (Chemistry of Natural Compounds Department, National Research Centre) ;
  • Kotob, Soheir E (Hormones Department, National Research Centre)
  • Published : 2015.11.04


Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.


Boswellia serrata;phytochemical analysis;HepG2 cell line;HCT 116 cell line;cytotoxicity


  1. Abd-Rabou AA, Zoheir KM, Ahmed HA (2012). Potential impact of curcumin and taurine on human hepatoma cells using Huh-7 cell line. Clinical Biochem, 45, 1519-21.
  2. Adams RP (1995). Identification of essential oil components by GC/MS. allured publ. Co, carol stream.
  3. Aggarwal BB, Shishodia S (2006). Molecular targets of dietary agents for prevention and therapy of cancer. Biochem Pharmacol, 71, 1397-421.
  4. Alam M, Khan H, Samiullah L, et al (2012). A review on phytochemical and pharmacological studies of kundur (boswellia serrata roxb ex colebr.) -a unani drug. J Applied Pharmaceutical Sci, 2, 148-56.
  5. Ali NAA, Wurster M, Arnold N, et al (2008). Chemical composition and biological activities of essential oils from the oleogum resins of three endemic soqotraen boswellia species, Rec Nat Prod, 2, 6-12.
  6. Ammon HPT, Safayhi H, Mark T, et al (1993). echanism of antiinflammatory actions of curcumine and boswellic acids. J Ethnopharmacol, 38, 105-12.
  7. Armitage P, Berry G (1987). Comparison of several groups. In, statistical method in medical research 2nd Ed. blockwell significant publication, Oxford. pp. 186-213.
  8. Arponsuwan T, Punjanon T (2006). Tumor cellselective antiproliferative effect of the extract from Morindacitrifoliafruits. Phytotherapy Res, 20, 515-7.
  9. Bhagat J, Lobo R, Kumar N, et al (2014). Cytotoxic potential of Anisochiluscarnosus (L.f.) wall and estimation of luteolin content by HPLC. BMC Complement Alternative Med, 14, 421.
  10. Bhushan S, Kumar A, Malik F, et al (2007). A triterpenediol from Boswellia serrata induces apoptosis through both the intrinsic and extrinsic apoptotic pathways in human leukemia HL-60 cells. Apoptosis, 12, 1911-26
  11. Bishayee A (2012). Editorial, recent advances in the prevention and therapy of hepatocellular carcinoma. Current Cancer Drug Targets, 12, 1043-4.
  12. Center MM, Jemal A (2011). International trends in liver cancer incidence rates. Cancer Epidemiol Biomarkers Prevention, 20, 2362-8.
  13. Demir T, Ozen MO, Hames-Kocabas EE (2015). Antioxidant and cytotoxic activity of Physalisperuviana. Medicinal Plant Research, 4, 30-34.
  14. El-Serag HB, Rudolph KL (2007). Hepatocellular carcinoma, epidemiology and molecular carcinogenesis. Gastroenterol, 132, 2557-76.
  15. Fahraeus R, Paramio JM, Ball KL, et al (1996). Inhibition of pRb phosphorylation and cell-cycle progression by a 20-residue peptide derived from p16. Curr Biol, 6, 84-91.
  16. Finar IL (1967). Organic chemistry. 5th ed . London, Longmans Green and Co. Ltd, 1, 212.
  17. Frank MB, Yang Q, Osban J, et al (2009). Frankincense oil derived from Boswellia carteriinduces tumor cell specific cytotoxicity. BMC Complement Altern. Med, 9, 6.
  18. Gao J, Xie L, Yang WS (2012). Risk factors of hepatocellular carcinoma current status and perspectives. Asian Pac J Cancer Prev, 13, 743-52.
  19. Godefroot M, Sandra P, Verzele M (1981). New method for quantitative essential oil analysis. J Chrom, 203, 325-35.
  20. Hayashi Y, Nishikawa Y, Mori H, et al (1998). Antitumor activity of (1 OE, 122)-9-hydroxy- 10,12-octadecadienoic acid from rice bran. J Fermentat Bioengineer, 86, 149-53.
  21. Hostanska K, Daum G, Saller R (2002). Cytostatic and apoptosisinducing activity of boswellic acids toward malignant cell lines in vitro. Anticancer Res, 22, 2853-62.
  22. Huang MT, Badmaev V, Ding Y, et al (2000). Anti-tumor and anti-carcinogenic activities of triterpenoid,${\beta}$-boswellic acid. Bio Factors, 13, 225-30.
  23. Kawanishi S, Hiraku Y, Pinlaor S, et al (2006). Oxidative and nitrative DNA damage in animals and patients with inflammatory diseases in relation to inflammation related carcinogenesis. Biological Chemist, 387, 365-72.
  24. Kim SJ, Park KM, Kim N, et al (2006). Doxorubicin prevents endoplasmic reticulum stress-induced apoptosis. Biochem Biophys Res Commun, 339, 463-8.
  25. Kumar A, Shah BA, Singh S, et al (2012). Acyl derivatives of boswellic acids as inhibitors of NF-kB and STATs. Bioorg Med Chem Lett, 22, 431-5.
  26. Kumar MS, Pal AK (2012). Investigation of bioactivity of extracts of marine sponge, spongosoriteshalichondrioides (dendy, 1905) from western coastal areas of India. Asian Pac J Tropical Biomedicine, 1784-9.
  27. Ley R, Ewings KE, Hadfield K, et al (2005). Regulatory phosphorylation of Bim, sorting out the ERK from the JNK. Cell Death Differ., 12(8), 1008-14.
  28. Lin J, Dou J, Xu J, (2012). Chemical composition, antimicrobial and antitumor activities of the essential oils and crude extracts of euphorbia macrorrhiza. Molecules, 17, 5030-9.
  29. Lin HK, Suhail MM, Fung KM, et al (2013). Extraction of biologically active compounds by hydrodistillation of Boswellia species gum resins for anti-cancer therapy. OA Alternat Med, 1, 4.
  30. Liu JJ, Huang B, Hooi SC (2006). Acetyl-keto-${\beta}$-boswellic acid inhibits cellular proliferation through a p21-dependent pathway in colon cancer cells. Br J Pharmacol, 148, 1099-107.
  31. Liu JJ, Nilsson A, Oredsson S, et al (2002). 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, 23, 2087-93.
  32. Lu M, Xia L, Hua H, et al (2008). Acetyl-keto-${\beta}$-boswellic acid induces apoptosis through a death receptor 5-mediated pathway in prostate cancer cells. Cancer Res, 68, 1180-6.
  33. Mitupatum T, Aree K, Kittisenachai S, et al (2015). Hep88 mAb-mediated paraptosis-like apoptosis in HepG2 cells via downstream upregulation and activation of caspase-3, caspase-8 and caspase-9. Asian Pac J Cancer Prev, 16, 1771-9
  34. Ni X, Suhail MM, Yang Q, et al (2012). Frankincense essential oil prepared from hydrodistillation of sacra gum resins induces human pancreatic cancer cell death in cultures and in a xenograft murine model. BMC Complement Altern. Med, 12, 253.
  35. Panaretakis T, Laane E, Pokrovskaja K, et al (2005). Doxorubicin requires the sequential activation of caspase-2, protein kinase Cdelta, and c-Jun NH2-terminal kinase to induce apoptosis. Mol Biol Cell, 16, 3821-31.
  36. Pandey RS, Singh BK, Tripathi YB (2005). Extract of gum resins of Boswellia serrata L. inhibits lipopolysaccharide induced nitric oxide production in rat macrophages along with hypolipidemic property. Indian J Experimental Biol, 43, 509-16.
  37. Park B, Sung B, Yadav VR, et al (2011). Acetyl-11-keto-bboswellic acid suppresses invasion of pancreatic cancer cells through the downregulation of CXCR4 chemokine receptor expression. Int J Cancer, 129, 23-33.
  38. Park YS, Lee JH, Bondar J, et al (2002). Cytotoxic action of acetyl-11-keto-b-boswellic acid (AKBA) on meningioma cells. Planta Med, 68, 397-401.
  39. Puthalakath H, O'Reilly LA, Gunn P, et al (2007). ER stress triggers apoptosis by activating BH3-only protein Bim. Cell, 129, 1337-49.
  40. Quelle DE, Ashmun RA, Shurtleff SA, et al (1993). Overexpression of mouse D-type cyclins accelerates G1 phase in rodent fibroblasts. Genes Dev, 7, 1559-71.
  41. Rang HP, Dale MM, Ritter JM, et al (2003). Book review, pharmacology. London, churchill livingstone. fifth edition.
  42. Roussi S, Winter A, Gosse F, et al (2005). Different apoptotic mechanisms are involved in the antiproliferative effects of 7${\beta}$-hydroxysitosterol and 7${\beta}$-hydroxycholesterol in human colon cancer cells. Cell Death Differentiat, 12, 128-35.
  43. Safayhi H, Sailer ER (1997). Anti-inflammatory actions of pentacyclictriterpenes. Planta Med, 63, 487-93.
  44. Schnekenburger M, Diederich M (2012). Epigenetics offer new horizons for colorectal cancer prevention. Curr Colorectal Cancer Rep, 8, 66-81.
  45. Shah BA, Qazi GN, Taneja SC (2009). Boswellic acids, a group of medicinally important compounds. Nat Prod Rep, 26, 72-89
  46. Shahat AA, Alsaid MS, Kotob SE, et al (2015). Significance of rumex vesicarius as anticancer remedy against hepatocellular carcinoma, a proposal-based on experimental animal studies. Asian Pac J Cancer Prev, 16, 4303-10
  47. Shao Y, Ho CT, Chin CK, et al (1998). Inhibitory activity of boswellic acids from Boswellia serrata against human leukemia HL-60 cells in culture. Planta Med, 64, 328-31.
  48. Sharma A, Chhikara S, Ghodekar SN, et al (2009). Phytochemical and pharmacological investigations on boswellia serrate. Phcog Rev, 3, 206-215.
  49. Skehan P, Storeng R, Scudiero D, et al (1990). New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst, 82, 1107-12.
  50. Suhail MM, Wu W, Cao A, et al (2011). Boswellia sacra essential oil induces tumor cell-specific apoptosis and suppresses tumoraggressiveness in cultured human breast cancer cells. BMC Complement Altern Med, 11, 129.
  51. Syrovets T, Gschwend JE, Buchele B, et al (2005). 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, 280, 6170-80.
  52. Tanaka S, Arii S (2009). Molecularly targeted therapy for hepatocellular carcinoma. Cancer Sci, 100, 1-8.
  53. Thomas MB, Jaffe D, Chotietal MM (2010). Hepatocellular carcinoma, consensus recommendations of the national cancer institute clinical trials planning meeting. Clin Oncol, 28, 3994-4005
  54. Tsuda K, Sakai K, Tanbe K, et al (1960). Isolation of 22-dehydrocholesterol from hypnea japonica. J Am Chem Society, 82, 1442.
  55. Uddin SJ, Grice ID, Tiralongo E (2009). Cytotoxic effects of bangladeshi medicinal plant extracts. evid. based complement. Alternat Med, 111, 1-6.
  56. Watson AJ, Collins PD (2011). Colon cancer, a civilization disorder. Digestive Disease, 29, 222-8.
  57. Winking M, Sarikaya S, Rahmanian A, et al (2000). Boswellic acids inhibit glioma growth, a new treatment option? J Neurooncol, 46, 97-103.
  58. World Cancer Research Fund and American Institute for Cancer Research (1997). Food, nutrition and the prevention of cancer, a global perspective, american institute for cancer research, washington, dc., pp. 1-670.
  59. wu sj, ng lt, chen CH, et al (2004). Antihepatoma activity of Physalisangulata and P. peruviana extracts and their effects on apoptosis in human Hep G2 cells. Life Sci, 74, 2061-73.
  60. Yazdanpanahi N, Behbahani M, Yektaeian A (2014). Effect of boswellia thurifera gum methanol extract on cytotoxicity and P53 gene expression in human breast cancer cell line, Iranian J Pharmaceutical Res, 13, 719-24.
  61. Zhang N, Yin Y, Xu S, et al (2008). 5-Fluorouracil, mechanisms of resistance and reversal strategies. Molecules, 13, 1551-69.

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