β-sitosterol induces reactive oxygen species-mediated apoptosis in human hepatocellular carcinoma cell line | ||
Avicenna Journal of Phytomedicine | ||
دوره 11، شماره 6، بهمن و اسفند 2021، صفحه 541-550 اصل مقاله (577.89 K) | ||
نوع مقاله: Short communication | ||
شناسه دیجیتال (DOI): 10.22038/ajp.2021.17746 | ||
نویسندگان | ||
Devaraj Ezhilarasan* ؛ Mary J Ditty | ||
Department of Pharmacology, The Blue Lab, Molecular Medicine and Toxicology Division, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Chennai, Tamil Nadu, India | ||
چکیده | ||
Objective: It is of interest to investigate the anti-proliferative effect of β-sitosterol (BS) on human hepatocellular carcinoma (HepG2) cell line. Materials and Methods: β-sitosterol treatments (0.6 and 1.2 mM/ml) were done in HepG2 and after 24 hr, cell viability was evaluated by MTT assay. Reactive oxygen species (ROS) accumulating potential of BS was assessed by dichloro-dihydro-fluorescein diacetate staining. Morphology related to apoptosis was investigated by acridine orange and ethidium bromide dual staining. Cytochrome c and caspase 3 expressions were evaluated by immunofluorescence and western blot analyses. Results: β-sitosterol induced cytotoxicity (p<0.001) and intracellular ROS in HepG2 cells in a dose-dependent manner.BS treatments accumulated induced intracellular ROS accumulation which led to membrane damage and mitochondrial toxicity. At the molecular level, BS treatments induced cytochrome c release from mitochondria and enhanced the protein expressions (p<0.05 vs 0.6 mM/ml and p<0.001 vs 1.2 mM/ml) of both caspase 3 and cleaved caspase 3. Conclusion: β-sitosterol induced ROS accumulation which plays a critical role in apoptosis via the intrinsic pathway in HepG2 cells. The present investigation paves the way for further in vivo studies. | ||
کلیدواژهها | ||
Liver cancer؛ β-sitosterol؛ Reactive oxygen species؛ Apoptosis؛ Caspase | ||
مراجع | ||
Alvarez-Sala A, Attanzio A, Tesoriere L, Garcia-Llatas G, Barberá R, Cilla A. 2019. Apoptotic effect of a phytosterol-ingredient and its main phytosterol (β-sitosterol) in human cancer cell lines. Int J Food Sci Nutr, 70: 323-334. Solai Prakash AK, Devaraj E. 2019. Cytotoxic potentials of S. cumini methanolic seed kernel extract in human hepatoma HepG2 cells. Environ Toxicol, 34: 1313-1319. Awad AB, Barta SL, Fink CS, Bradford PG. 2008. Beta-sitosterol enhances tamoxifen effectiveness on breast cancer cells by affecting ceramide metabolism. Mol Nutr Food Res, 52: 419-426. Awad AB, Fink CS, Williams H, Kim U. 2001. In vitro and in vivo (SCID mice) effects of phytosterols on the growth and dissemination of human prostate cancer PC3 cells. Eur J Cancer Prev, 10: 507-513. Bacchetti T, Masciangelo S, Bicchiega V, Bertoli E, Ferretti G. 2011. Phytosterols, phytostanols and their esters: from natural to functional foods. Med J Nutrition Metab, 4: 165–172. Baskar AA, Al Numair KS, Gabriel Paulraj M, Alsaif MA, Muamar MA, Ignacimuthu S. 2012. β-sitosterol prevents lipid peroxidation and improves antioxidant status and histoarchitecture in rats with 1,2- dimethylhydrazine-induced colon cancer. J Med Food, 15: 335-343. Baskar AA, Ignacimuthu S, Paulraj GM, Al Numair KS. 2010. Chemopreventive potential of beta-Sitosterol in experimental colon cancer model-an in vitro and In vivo study. BMC Complement Altern Med, 10: 24. Byczkowska A, Kunikowska A, Kaźmierczak A. 2013. Determination of ACC-induced cell-programmed death in roots of Vicia faba ssp. minor seedlings by acridine orange and ethidium bromide staining. Protoplasma, 250: 121-128. Cao ZQ, Wang XX, Lu L, Xu JW, Li XB, Zhang GR, Ma ZJ, Shi AC, Wang Y, Song YJ. 2019. β-sitosterol and gemcitabine exhibit synergistic anti-pancreatic cancer activity by modulating apoptosis and inhibiting epithelial-mesenchymal transition by deactivating Akt/GSK-3β signaling. Front Pharmacol, 9: 1525. Chirumbolo S, Bjørklund G, Lysiuk R, Vella A, Lenchyk L, Upyr T. 2018. Targeting cancer with phytochemicals via their fine tuning of the cell survival signaling pathways. Int J Mol Sci, 19: 3568. Choi YH, Kong KR, Kim YA, Jung KO, Kil JH, Rhee SH, Park KY. 2003. Induction of Bax and activation of caspases during betasitosterol-mediated apoptosis in human colon cancer cells. Int J Oncol, 23: 1657- 1662. Devaraj E, Roy A, Royapuram Veeraragavan G, Magesh A, Varikalam Sleeba A, Arivarasu L, Marimuthu Parasuraman B. 2020. β-Sitosterol attenuates carbon tetrachloride-induced oxidative stress and chronic liver injury in rats. Naunyn Schmiedebergs Arch Pharmacol, 393: 1067- 1075. Ezhilarasan D, Apoorva VS, Ashok Vardhan N. 2019. Syzygium cumini extract induced reactive oxygen species-mediated apoptosis in human oral squamous carcinoma cells. J Oral Pathol Med, 48: 115-121. Ezhilarasan D. 2018. Herbal therapy for cancer. In: Prakash Srinivasan, Timiri Shanmugam., editors. Understanding Cancer Therapies, CRC Press, pp. 129–166. Gheena S, Ezhilarasan D. 2019. Syringic acid triggers reactive oxygen species-mediated cytotoxicity in HepG2 cells. Hum Exp Toxicol, 38: 694-702. Hartke J, Johnson M, Ghabril M. 2017. The diagnosis and treatment of hepatocellular carcinoma. Semin Diagn Pathol, 34: 153- 159. Hong SH, Cha HJ, Hwang-Bo H, Kim MY, Kim SY, Ji SY, Cheong J, Park C, Lee H, Kim GY, Moon SK, Yun SJ, Chang YC, Kim WJ, Choi YH. 2019. Anti-proliferative and pro-apoptotic effects of licochalcone A through ROS-mediated cell cycle arrest and apoptosis in human bladder cancer cells. Int J Mol Sci, 20: 3820. Hosseini A, Ghorbani A. 2015. Cancer therapy with phytochemicals: evidence from clinical studies. Avicenna J Phytomed, 5: 84-97. Jan R, Chaudhry GE. 2019. Understanding apoptosis and apoptotic pathways targeted cancer therapeutics. Adv Pharm Bull, 9: 205-218. Jiang JF, Lao YC, Yuan BH, Yin J, Liu X, Chen L, Zhong JH. 2017. Treatment of hepatocellular carcinoma with portal vein tumor thrombus: advances and challenges. Oncotarget, 8: 33911-33921. β-sitosterol induces cytotoxicity in HepG2 cells AJP, Vol. 11, No. 6, Nov-Dec 2021 549 Jiang L, Zhao X, Xu J, Li C, Yu Y, Wang W, Zhu L. 2019. The protective effect of dietary Phytosterols on cancer risk: A systematic meta-analysis. J Oncol, 2019: 7479518. Jones PJ, AbuMweis SS. 2009. Phytosterols as functional food ingredients: linkages to cardiovascular disease and cancer. Curr Opin Clin Nutr Metab Care, 12: 147-151. Kasibhatla S, Amarante-Mendes GP, Finucane D, Brunner T, Bossy-Wetzel E, Green DR. 2006. Acridine orange/ethidium bromide (AO/EB) staining to detect apoptosis. CSH Protoc, 2006: pdb.prot4493. Kazłowska K, Lin HT, Chang SH, Tsai GJ. 2013. In vitro and in vivo anticancer effects of sterol fraction from red algae porphyra dentata. Evid Based Complement Alternat Med, 2013: 493869. Lakshmi T, Ezhilarasan D, Vijayaragavan R, Bhullar SK, Rajendran R. 2017. Acacia catechu ethanolic bark extract induces apoptosis in human oral squamous carcinoma cells. J Adv Pharm Technol Res, 8: 143-149. Li S, Jiang Z, Chai W, Xu Y, Wang Y. 2019. Autophagy activation alleviates nonylphenol-induced apoptosis in cultured cortical neurons. Neurochem Int, 122: 73- 84. Li X, Wu Q, Bu M, Hu L, Du WW, Jiao C, Pan H, Sdiri M, Wu N, Xie Y, Yang BB. 2016. Ergosterol peroxide activates Foxo3- mediated cell death signaling by inhibiting AKT and c-Myc in human hepatocellular carcinoma cells. Oncotarget, 7: 33948- 33959. Liu K, Liu PC, Liu R, Wu X. 2015. Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry. Med Sci Monit Basic Res, 21: 15-20. Misawa E, Tanaka M, Nomaguchi K, Nabeshima K, Yamada M, Toida T, Iwatsuki K. 2012. Oral ingestion of aloe vera phytosterols alters hepatic gene expression profiles and ameliorates obesityassociated metabolic disorders in zucker diabetic fatty rats. J Agric Food Chem, 60: 2799-2806. Pfeffer CM, Singh ATK. 2018. Apoptosis: A target for anticancer therapy. Int J Mol Sci, 19: E448. PonselviInduja M, Ezhilarasan D, Ashok Vardhan N. 2018. Evolvulus alsinoides methanolic extract triggers apoptosis in HepG2 cells. Avicenna J Phytomed, 8: 504- 512. Kathiswar R, Ezhilarasan D, Rajeshkumar S. 2020. β-Sitosterol-assisted silver nanoparticles activates Nrf2 and triggers mitochondrial apoptosis via oxidative stress in human hepatocellular cancer cell line. J Biomed Mater Res A, 108: 1899-1908. Rajavel T, Packiyaraj P, Suryanarayanan V, Singh SK, Ruckmani K, Pandima Devi K. 2018. β-sitosterol targets Trx/Trx1 reductase to induce apoptosis in A549 cells via ROS mediated mitochondrial dysregulation and p53 activation. Sci Rep, 8: 2071. Rajavel T, Mohankumar R, Archunan G, Ruckmani K, Devi KP. 2017. Beta sitosterol and Daucosterol (phytosterols identified in Grewia tiliaefolia) perturbs cell cycle and induces apoptotic cell death in A549 cells. Sci Rep, 7: 3418. Ramprasath VR, Awad AB. 2015. Role of phytosterols in cancer prevention and treatment. J AOAC Int, 98: 735-738. Rayan A, Raiyn J, Falah M. 2017. Nature is the best source of anticancer drugs: Indexing natural products for their anticancer bioactivity. PLoS One, 12: e0187925. Rohit Singh T, Ezhilarasan D. 2019. Ethanolic extract of Lagerstroemia Speciosa (L.) Pers., induces apoptosis and cell cycle arrest in HepG2 cells. Nutr Cancer, 72: 146-156. Sharmila R, Sindhu G. 2017. Modulation of angiogenesis, proliferative response and apoptosis by BS in rat model of renal carcinogenesis. Indian J Clin Biochem, 32: 142-152. Shebi S, Ezhilarasan D, Thomas J, Chandrasekaran N, Mukherjee A. 2019. Gracilaria foliifera (Forssk.) Børgesen ethanolic extract triggers apoptosis via activation of p53 expression in HepG2 cells. Phcog Mag, 15: 259-263. Shin EJ, Choi HK, Sung MJ, Park JH, Chung MY, Chung S, Hwang JT. 2018. Antitumour effects of beta-sitosterol are mediated by AMPK/PTEN/HSP90 axis in AGS human gastric adenocarcinoma cells and xenograft mouse models. Biochem Pharmacol, 152: 60-70. Siegel RL, Miller KD, Jemal A. 2016. Cancer statistics, 2016.CA Cancer J Clin, 66: 7-30. Suh JK, Lee J, Lee JH, Shin S, Tchoe HJ, Kwon JW. 2018. Risk factors for developing liver cancer in people with and without liver disease. PLoS One, 13: e0206374. Ditty et al. AJP, Vol. 11, No. 6, Nov-Dec 2021 550 Suttiarporn P, Chumpolsri W, Mahatheeranont S, Luangkamin S, Teepsawang S, Leardkamolkarn V. 2015. Structures of phytosterols and triterpenoids with potential anti-cancer activity in bran of black nonglutinous rice. Nutrients, 7: 1672-1687. Yang C, Chen ZY, Wong SL, Liu J, Liang YT, Lau CW, Lee HK, Huang Y, Tsang SY. 2013. β-Sitosterol oxidation products attenuate vasorelaxation by increasing reactive oxygen species and cyclooxygenase-2. Cardiovasc Res, 97: 520- 532. Yang Y, Karakhanova S, Hartwig W, D'Haese JG, Philippov PP, Werner J, Bazhin AV. 2016. Mitochondria and mitochondrial ROS in cancer: Novel targets for anticancer therapy. J Cell Physiol, 231: 2570-2581. Zaloga GP. 2015. Phytosterols, lipid administration, and liver disease during parenteral nutrition. JPEN J Parenter Enteral Nutr, 39: 39S-60S. | ||
آمار تعداد مشاهده مقاله: 18,064 تعداد دریافت فایل اصل مقاله: 814 |