In vivo attenuation of angiogenesis in hepatocellular carcinoma by Nigella sativa

Background/aim: Angiogenesis is imperative in malignant tumor growth. Hepatocellular carcinoma is a typical hypervasculartumor that relies on angiogenesis. The aim of this study is to investigate the in vivo molecular mechanism underlying the antitumorproperties of Nigella sativa ethanolic extract (NSEE) through its antiangiogenic effect against diethyl nitrosamine (DENA)-inducedhepatocarcinogenesis.Materials and methods: Male Wistar rats were divided into 4 groups: normal, NSEE, DENA, and NSEE-DENA groups. Final bodyweight, hepatosomatic indices, serum AFP, serum vascular endothelial growth factor (VEGF) levels, and liver tissue hepatocyte growthfactor β (HGFβ) protein expression were estimated. Liver sections were stained for histological examination. AFP levels with thehistological variations were chosen to confirm cancer development.Results: DENA significantly increased liver weight, hepatosomatic indices, serum AFP and VEGF levels, and liver HGFβ proteinexpression and significantly decreased final body weight. These effects were significantly reversed by NSEE. Furthermore, thehistopathological changes that appeared in rat livers due to DENA were reduced by NSEE without harmful effects when taken alone.Conclusion:The results of the present investigation provide evidence that the in vivo antiangiogenic effect of NSEE through downregulationof serum VEGF and liver HGFβ protein could be implicated in its antitumor activity. Its consumption has health benefits that favor livercancer management. These findings might prove useful and helpful for the progression of therapies for hepatocarcinogenesis treatment.

PDF

___

Siveen KS, Prabhu K, Krishnankutty R, Kuttikrishnan S, Tsakou M, Alali FQ, Dermime S, Mohammad RM, Uddin S. Vascular endothelial growth factor (VEGF) signaling in tumour vascularization: potential and challenges. Curr Vasc Pharmacol 2017; 15: 339-351.
Mueller MM, Fusenig NE. Friends or foes – bipolar effects of the tumour stroma in cancer. Nat Rev Cancer 2004; 4: 839-849.
Budhu A, Forgues M, Ye QH, Jia HL, He P, Zanetti KA, Kammula US, Chen Y, Qin LX, Tang ZY et al. Prediction of venous metastases, recurrence, and prognosis in hepatocellular carcinoma based on a unique immune response signature of the liver microenvironment. Cancer Cell 2006; 10: 99-111.
Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors. Nat Med 2003; 9: 669-676.
Grothey A, Galanis E. Targeting angiogenesis: progress with anti-VEGF treatment with large molecules. Nat Rev Clin Oncol 2009; 6: 507-518.
Gonzalez-Moreno O, Lecanda J, Green JE, Segura V, Catena R, Serrano D, Calvo A. VEGF elicits epithelial-mesenchymal transition (EMT) in prostate intraepithelial neoplasia (PIN)- like cells via an autocrine loop. Exp Cell Res 2010; 316: 554- 567.
Fujiuchi Y, Nagakawa O, Murakami K, Fuse H, Saiki I. Effect of hepatocyte growth factor on invasion of prostate cancer cell lines. Oncol Rep 2003; 10: 1001-1006.
Sengupta S, Sellers LA, Cindrova T, Skepper J, Gherardi E, Sasisekharan R, Fan TP. Cyclooxygenase-2-selective nonsteroidal anti-inflammatory drugs inhibit hepatocyte growth factor/scatter factor-induced angiogenesis. Cancer Res 2003; 63: 8351-8359.
Finn RS. Development of molecularly targeted therapies in hepatocellular carcinoma: where do we go now? Clin Cancer Res 2010; 16: 390-397.
Parkin DM, Bray F, Ferlay J, Pisani P. Estimating the world cancer burden: GLOBOCAN 2000. Int J Cancer 2001; 94: 153- 156.
Zhu AX. Systemic therapy of advanced hepatocellular carcinoma: how hopeful should we be? Oncologist 2006; 11: 790-800.
Semenza GL. Angiogenesis in ischemic and neoplastic disorders. Ann Rev Med 2003; 54: 17-28.
Jung JO, Gwak GY, Lim YS, Kim CY, Lee HS. Role of hepatic stellate cells in the angiogenesis of hepatoma. Korean J Gastroenterol 2003; 42: 142-148 (in Korean with an abstract in English).
Hu X, Li S, He Y, Ai P, Wu S, Su Y, Li X, Cai L, Peng X. Antitumor and antimetastatic activities of a novel benzothiazole-2-thiol derivative in a murine model of breast cancer. Oncotarget 2017; 8: 11887-11895.
Manzo A, Carillio G, Montanino A, Costanzo R, Sandomenico C, Rocco G, Morabito A. Focus on nintedanib in NSCLC and other tumors. Front Med 2016; 3: 68-78.
Abdel-Hamid NM, El-Moselhy MA, El-Baz A. Hepatocyte lysosomal membrane stabilization by olive leaves against chemically induced hepatocellular neoplasia in rats. Int J Hepatol 2011; 2011: 736581.
Randhawa MA, Alghamdi MS. Anticancer activity of Nigella sativa (black seed): a review. Am J Chin Med 2011; 39: 1075- 1091.
Ashraf SS, Rao MV, Kaneez FS, Qadri S, Al-Marzouqi AH, Chandranath IS, Adem A. Nigella sativa extract as a potent antioxidant for petrochemical-induced oxidative stress. J Chromatogr Sci 2011; 49: 321-326.
Kensara OA, El-Shemi AG, Mohamed AM, Refaat B, Idris S, Ahmad J. Thymoquinone subdues tumor growth and potentiates the chemopreventive effect of 5-fluorouracil on the early stages of colorectal carcinogenesis in rats. Drug Des Devel Ther 2016; 10: 2239-2253.
Ali BH, Blunden G. Pharmacological and toxicological properties of Nigella sativa. Phytother Res 2003; 17: 299-305.
Salem ML. Immunomodulatory and therapeutic properties of the Nigella sativa L. seed. Int Immunopharmacol 2005; 5: 1749- 1770.
Padhye S, Banerjee S, Ahmad A, Mohammad R, Sarkar FH. From here to eternity – the secret of pharaohs: therapeutic potential of black cumin seeds and beyond. Cancer Ther 2008; 6: 495-510.
Ong YS, Saiful Yazan L, Ng WK, Noordin MM, Sapuan S, Foo JB, Tor YS. Acute and subacute toxicity profiles of thymoquinone-loaded nanostructured lipid carrier in BALB/c mice. Int J Nanomed 2016; 11: 5905-5915.
Badr G, Mohany M, Abu-Tarboush F. Thymoquinone decreases F-actin polymerization and the proliferation of human multiple myeloma cells by suppressing STAT3 phosphorylation and Bcl2/Bcl-XL expression. Lipids Health Dis 2011; 10: 236.
Banerjee S, Padhye S, Azmi A, Wang Z, Philip PA, Kucuk O, Sarkar FH, Mohammad RM. Review on molecular and therapeutic potential of thymoquinone in cancer. Nutr Cancer 2010; 62: 938-946.
Jafri SH, Glass J, Shi R, Zhang S, Prince M, Kleiner-Hancock H. Thymoquinone and cisplatin as a therapeutic combination in lung cancer: in vitro and in vivo. J Exp Clin Cancer Res 2010; 29: 87.
Kolli-Bouhafs K, Boukhari A, Abusnina A, Velot E, Gies JP, Lugnier C, Ronde P. Thymoquinone reduces migration and invasion of human glioblastoma cells associated with FAK, MMP-2 and MMP-9 down-regulation. Invest New Drugs 2011; 30: 2121-2131.
Lei X, Lv X, Liu M, Yang Z, Ji M, Guo X, Dong W. Thymoquinone inhibits growth and augments 5-fluorouracilinduced apoptosis in gastric cancer cells both in vitro and in vivo. Biochem Biophys Res Commun 2012; 417: 864-868.
Sethi G, Ahn KS, Aggarwal BB. Targeting nuclear factor-kappa B activation pathway by thymoquinone: role in suppression of antiapoptotic gene products and enhancement of apoptosis. Mol Cancer Res 2008; 6: 1059-1070.
Torres MP, Ponnusamy MP, Chakraborty S, Smith LM, Das S, Arafat HA, Batra SK. Effects of thymoquinone in the expression of mucin 4 in pancreatic cancer cells: implications for the development of novel cancer therapies. Mol Cancer Ther 2010; 9: 1419-1431.
Woo CC, Kumar AP, Sethi G, Tan KH. Thymoquinone: potential cure for inflammatory disorders and cancer. Biochem Pharmacol 2012; 83: 443-451.
Fathy M, Nikaido T. In vivo modulation of iNOS pathway in hepatocellular carcinoma by Nigella sativa. Environ Health Prev Med 2013; 18: 377-385.
Michel CG, El-Sayed NS, Moustafa SF, Ezzat SM, Nesseem DI, El-Alfy TS. Phytochemical and biological investigation of the extracts of Nigella sativa L. seed waste. Drug Test Anal 2011; 3: 245-254.
Sayed-Ahmed MM, Aleisa AM, Al-Rejaie SS, Al-Yahya AA, AlShabanah OA, Hafez MM, Nagi MN. Thymoquinone attenuates diethylnitrosamine induction of hepatic carcinogenesis through antioxidant signaling. Oxid Med Cell Longev 2010; 3: 254-261.
Carmeliet P, Jain RK. Angiogenesis in cancer and other diseases. Nature 2000; 407: 249-257.
Streubel B, Chott A, Huber D, Exner M, Jager U, Wagner O, Schwarzinger I. Lymphoma-specific genetic aberrations in microvascular endothelial cells in B-cell lymphomas. N Engl J Med 2004; 351: 250-259.
Aznavoorian S, Murphy AN, Stetler-Stevenson WG, Liotta LA. Molecular aspects of tumor cell invasion and metastasis. Cancer 1993; 71: 1368-1383.
Li CX, Shao Y, Ng KT, Liu XB, Ling CC, Ma YY, Geng W, Fan ST, Lo CM, Man K. FTY720 suppresses liver tumor metastasis by reducing the population of circulating endothelial progenitor cells. PLoS One 2012; 7: e32380.
Lin Y, Zhai E, Liao B, Xu L, Zhang X, Peng S, He Y, Cai S, Zeng Z, Chen M. Autocrine VEGF signaling promotes cell proliferation through a PLC-dependent pathway and modulates apatinib treatment efficacy in gastric cancer. Oncotarget 2017; 8: 11990- 12002.
Ribatti D, Vacca A, Nico B, Sansonno D, Dammacco F. Angiogenesis and anti-angiogenesis in hepatocellular carcinoma. Cancer Treat Rev 2006; 32: 437-444.
Uematsu S, Higashi T, Nouso K, Kariyama K, Nakamura S, Suzuki M, Nakatsukasa H, Kobayashi Y, Hanafusa T, Tsuji T et al. Altered expression of vascular endothelial growth factor, fibroblast growth factor-2 and endostatin in patients with hepatocellular carcinoma. J Gastroenterol Hepatol 2005; 20: 583-588.
Montella L, Addeo R, Caraglia M, Faiola V, Guarrasi R, Vincenzi B, Palmeri A, Capasso E, Nocera V, Tarantino L et al. Vascular endothelial growth factor monitoring in advanced hepatocellular carcinoma patients treated with radiofrequency ablation plus octreotide: a single center experience. Oncol Rep 2008; 20: 385-390.
Tamesa T, Iizuka N, Mori N, Okada T, Takemoto N, Tangoku A, Oka M. High serum levels of vascular endothelial growth factor after hepatectomy are associated with poor prognosis in hepatocellular carcinoma. Hepatogastroenterology 2009; 56: 1122-1126.
Al-Ali A, Alkhawajah AA, Randhawa MA, Shaikh NA. Oral and intraperitoneal LD50 of thymoquinone, an active principle of Nigella sativa, in mice and rats. J Ayub Med Coll Abbottabad 2008; 20: 25-27.
Mansour MA, Ginawi OT, El-Hadiyah T, El-Khatib AS, AlShabanah OA, Al-Sawaf HA. Effects of volatile oil constituents of Nigella sativa on carbon tetrachloride-induced hepatotoxicity in mice: evidence for antioxidant effects of thymoquinone. Res Commun Mol Pathol Pharmacol 2001; 110: 239-251.
Iddamaldeniya SS, Thabrew MI, Wickramasinghe SM, Ratnatunge N, Thammitiyagodage MG. A long-term investigation of the anti-hepatocarcinogenic potential of an indigenous medicine comprised of Nigella sativa, Hemidesmus indicus and Smilax glabra. J Carcinog 2006; 5: 11.
Iddamaldeniya SS, Wickramasinghe N, Thabrew I, Ratnatunge N, Thammitiyagodage MG. Protection against diethylnitrosoamine-induced hepatocarcinogenesis by an indigenous medicine comprised of Nigella sativa, Hemidesmus indicus and Smilax glabra: a preliminary study. J Carcinog 2003; 2: 6.
Aggarwal BB, Shishodia S, Sandur SK, Pandey MK, Sethi G. Inflammation and cancer: how hot is the link? Biochem Pharmacol 2006; 72: 1605-1621.
Mansour MA, Al-Ismaeel H, Al-Rikabi AC, Al-Shabanah OA. Comparison of angiotensin converting enzyme inhibitors and angiotensin II type 1 receptor blockade for the prevention of premalignant changes in the liver. Life Sci 2011; 89: 188-194.
Singh BN, Singh BR, Sarma BK, Singh HB. Potential chemoprevention of N-nitrosodiethylamine-induced hepatocarcinogenesis by polyphenolics from Acacia nilotica bark. Chem Biol Interact 2009; 181: 20-28.
Taha MM, Abdul AB, Abdullah R, Ibrahim TA, Abdelwahab SI, Mohan S. Potential chemoprevention of diethylnitrosamineinitiated and 2-acetylaminofluorene-promoted hepatocarcinogenesis by zerumbone from the rhizomes of the subtropical ginger (Zingiber zerumbet). Chem Biol Interact 2010; 186: 295-305.
Majdalawieh AF, Hmaidan R, Carr RI. Nigella sativa modulates splenocyte proliferation, Th1/Th2 cytokine profile, macrophage function and NK anti-tumor activity. J Ethnopharmacol 2010; 131: 268-275.
Samarakoon SR, Thabrew I, Galhena PB, De Silva D, Tennekoon KH. A comparison of the cytotoxic potential of standardized aqueous and ethanolic extracts of a polyherbal mixture comprised of Nigella sativa (seeds), Hemidesmus indicus (roots) and Smilax glabra (rhizome). Pharmacogn Res 2010; 2: 335-342.
Alobaedi OH, Talib WH, Basheti IA. Antitumor effect of thymoquinone combined with resveratrol on mice transplanted with breast cancer. Asian Pac J Trop Med 2017; 10: 400-408.
Talib WH. Regressions of breast carcinoma syngraft following treatment with piperine in combination with thymoquinone. Sci Pharm 2017; 85: E27.
Poon RT, Ng IO, Lau C, Zhu LX, Yu WC, Lo CM, Fan ST, Wong J. Serum vascular endothelial growth factor predicts venous invasion in hepatocellular carcinoma: a prospective study. Ann Surg 2001; 233: 227-235.
Poon RT, Lau CP, Cheung ST, Yu WC, Fan ST. Quantitative correlation of serum levels and tumor expression of vascular endothelial growth factor in patients with hepatocellular carcinoma. Cancer Res 2003; 63: 3121-3126.
Reardon DA, Conrad CA, Cloughesy T, Prados MD, Friedman HS, Aldape KD, Mischel P, Xia J, Dilea C, Huang J et al. Phase I study of AEE788, a novel multitarget inhibitor of ErbBand VEGF-receptor-family tyrosine kinases, in recurrent glioblastoma patients. Cancer Chemother Pharmacol 2012; 69: 1507-1518.
Ma PC, Maulik G, Christensen J, Salgia R. c-Met: structure, functions and potential for therapeutic inhibition. Cancer Metast Rev 2003; 22: 309-325.
Reisinger K, Kaufmann R, Gille J. Increased Sp1 phosphorylation as a mechanism of hepatocyte growth factor (HGF/SF)-induced vascular endothelial growth factor (VEGF/ VPF) transcription. J Cell Sci 2003; 116: 225-238.
Van Belle E, Witzenbichler B, Chen D, Silver M, Chang L, Schwall R, Isner JM. Potentiated angiogenic effect of scatter factor/hepatocyte growth factor via induction of vascular endothelial growth factor: the case for paracrine amplification of angiogenesis. Circulation 1998; 97: 381-390.
Hendry SA, Farnsworth RH, Solomon B, Achen MG, Stacker SA, Fox SB. The role of the tumor vasculature in the host immune response: implications for therapeutic strategies targeting the tumor microenvironment. Front Immunol 2016; 7: 621.