Hypericin Exerts Detrimental Effect on Huh-7 As a Delegacy of Hepatocellular Carcinoma: A P53 Dependent Pathway
DOI:
https://doi.org/10.31661/gmj.v9i.1896Keywords:
Hypericin; Hepatocellular Carcinoma; Huh7; P53Abstract
Background: Hepatocellular carcinoma is the most common type of liver cancer which arises from the main cells in the liver. We address many studies investigating anti-cancer role of hypericin, however the proposing corresponding molecular pathway seems to be still a debate. Therefore, the present study aimed to evaluate the apoptotic effect of hypericin on the Huh7 as the liver cancer cell line and its relation with the gate keeper gene P53. Materials and Methods: In this study, the Huh7 cell line and fibroblast cells (as control group) were treated with different concentrations of hypericin for 24 and 48 hours. Detection of cell death was performed by MTT assay and flow cytometry. The expression of bax, bcl2 and p53 mRNAs was evaluated by Real-time PCR. Also, Immunocytochemistry (ICC) analysis was used for further evaluation of P53expression. Results: The results showed that hypericin has a dose-dependent cytotoxic effect on the Huh7 cell line, with no or marginal effect on fibroblastic cells. According to flow cytometry results, about 53%cells underwent apoptosis after exposure to LD50 of hypericin for 24 hours. Real-time PCR data demonstrated that the pro-apoptotic genes Bax and P53 expression level increased. Expectedly ICC results confirmed the up-regulation of P53 proteins in treated samples. Conclusion: Our results indicate the cytotoxicity of hypericin on Huh7 cells by affecting the expression of the gate keeper gene P53; furthermore it is suggested that this herb can be utilized simultaneously with modalities targeting P53 up-regulation or related molecular pathways. [GMJ.2020;9:e1896]
References
Zhao J, Fu Y, Wu J, Li J, Huang G, Qin L. The diverse mechanisms of miRNAs and lncRNAs in the maintenance of liver cancer stem cells. Biomed Res Int. 2018;2018. https://doi.org/10.1155/2018/8686027PMid:29888282 PMCid:PMC5977062 Shrestha A. Liver Cancer in Nepal. Euroasian J Hepatogastroenterol. 2018;8(1):63. https://doi.org/10.5005/jp-journals-10018-1261PMid:29963465 PMCid:PMC6024038 Rabinel P, Dousse D, Muscari F, Suc B. Management of liver cancer. The Surgeon's point of view. Rep Pract Oncol Radiother. 2017;22(2):176-80. https://doi.org/10.1016/j.rpor.2017.02.001PMid:28490990 PMCid:PMC5411896 Sia D, Villanueva A, Friedman SL, Llovet JM. Liver cancer cell of origin, molecular class, and effects on patient prognosis. Gastroenterology. 2017;152(4):745-61. https://doi.org/10.1053/j.gastro.2016.11.048PMid:28043904 Yu LX, Schwabe RF. The gut microbiome and liver cancer: mechanisms and clinical translation. Nat Rev Gastroenterol Hepatol. 2017;14(9):527. https://doi.org/10.1038/nrgastro.2017.72PMid:28676707 PMCid:PMC6467288 Mitchell JK, Lemon SM, McGivern DR. How do persistent infections with hepatitis C virus cause liver cancer? Curr Opin Virol. 2015;14:101-8. https://doi.org/10.1016/j.coviro.2015.09.003PMid:26426687 PMCid:PMC4628866 Bressac B, Puisieux A, Kew M, Volkmann M, Bozcall S, Mura JB et al. p53 mutation in hepatocellular carcinoma after aflatoxin exposure. Lancet. 1991;338(8779):1356-9. https://doi.org/10.1016/0140-6736(91)92236-U Abou-Alfa GK, Schwartz L, Ricci S, Amadori D, Santoro A, Figer A et al. Phase II study of sorafenib in patients with advanced hepatocellular carcinoma. J Clin Oncol. 2006;24(26):4293-300. https://doi.org/10.1200/JCO.2005.01.3441PMid:16908937 Farazi PA, DePinho RA. Hepatocellular carcinoma pathogenesis: from genes to environment. Nat Rev Cancer. 2006;6(9):674. https://doi.org/10.1038/nrc1934PMid:16929323 Wang Z, Li Z, Ye Y, Xie L, Li W. Oxidative stress and liver cancer: etiology and therapeutic targets. Oxid Med Cell Longev. 2016;2016. https://doi.org/10.1155/2016/7891574PMid:27957239 PMCid:PMC5121466 Gamasaee NA, Radmansouri M, Ghiasvand S, Shahriari F, Marzouni HZ, Aryan H et al. Hypericin induces apoptosis in MDA-MB-175-VII cells in lower dose compared to MDA-MB-231. Arch Iran Med. 2018;21(9):387-92. Ghiassi-Nejad Z, Tarchi P, Moshier E, Ru M, Tabrizian P, Schwartz M et al. Prognostic factors and patterns of Locoregional failure after surgical resection in patients with cholangiocarcinoma without adjuvant radiation therapy: optimal field Design for Adjuvant Radiation Therapy. Int J Radiat Oncol Biol Phys. 2017;99(4):805-11. https://doi.org/10.1016/j.ijrobp.2017.06.2467PMid:29063849 Bruix J, Reig M, Sherman M. Evidence-based diagnosis, staging, and treatment of patients with hepatocellular carcinoma. Gastroenterology. 2016;150(4):835-53. https://doi.org/10.1053/j.gastro.2015.12.041PMid:26795574 Javidi MA, Zolghadr F, Babashah S, Sadeghizadeh M. Introducing dendrosomal nanocurcumin as a compound capable of in vitro eliminating undifferentiated stem cells in cell therapy practices. Exp Clin Endocrinol Diabetes. 2015;123(10):632-6. https://doi.org/10.1055/s-0035-1555775PMid:26179929 Mirmalek SA, Jangholi E, Jafari M, Yadollah-Damavandi S, Javidi MA, Parsa Y et al. Comparison of in vitro cytotoxicity and apoptogenic activity of magnesium chloride and cisplatin as conventional chemotherapeutic agents in the MCF-7 cell line. Asian Pac J Cancer Prev. 2016;17(S3):131-4. https://doi.org/10.7314/APJCP.2016.17.S3.131PMid:27165250 Jaradat NA, Al-Ramahi R, Zaid AN, Ayesh OI, Eid AM. Ethnopharmacological survey of herbal remedies used for treatment of various types of cancer and their methods of preparations in the West Bank-Palestine. BMC Complement Altern Med. 2016;16(1):93. https://doi.org/10.1186/s12906-016-1070-8PMid:26955822 PMCid:PMC4784411 Ting CT, Kuo CJ, Hu HY, Lee YL, Tsai TH. Prescription frequency and patterns of Chinese herbal medicine for liver cancer patients in Taiwan: a cross-sectional analysis of the National Health Insurance Research Database. BMC Complement Altern Med. 2017;17(1):118. https://doi.org/10.1186/s12906-017-1628-0PMid:28219357 PMCid:PMC5319102 Kirakosyan A, Sirvent TM, Gibson DM, Kaufman PB. The production of hypericins and hyperforin by in vitro cultures of St. John's wort (Hypericum perforatum). Biotechnol Appl Biochem. 2004;39(1):71-81. https://doi.org/10.1042/BA20030144PMid:14521510 Wurglics M, Schubert-Zsilavecz M. Hypericum perforatum: A 'modern'herbal antidepressant. Clin Pharmacokinet. 2006;45(5):449-68. https://doi.org/10.2165/00003088-200645050-00002PMid:16640452 Zhai XJ, Chen F, Chen C, Zhu CR, Lu YN. LC-MS/MS based studies on the anti-depressant effect of hypericin in the chronic unpredictable mild stress rat model. J Ethnopharmacol. 2015;169:363-9. https://doi.org/10.1016/j.jep.2015.04.053PMid:25957811 Süntar I, Oyardı O, Akkol EK, Ozçelik B. Antimicrobial effect of the extracts from Hypericum perforatum against oral bacteria and biofilm formation. Pharm Biol. 2016;54(6):1065-70. https://doi.org/10.3109/13880209.2015.1102948PMid:26510970 Mirmalek SA, Azizi MA, Jangholi E, Yadollah-Damavandi S, Javidi MA, Parsa Y et al. Cytotoxic and apoptogenic effect of hypericin, the bioactive component of Hypericum perforatum on the MCF-7 human breast cancer cell line. Cancer Cell Int. 2015;16(1):3. https://doi.org/10.1186/s12935-016-0279-4PMid:26865836 PMCid:PMC4748624 Momekov G, Ferdinandov D, Zheleva-Dimitrova D, Nedialkov P, Girreser U, Kitanov G. Cytotoxic effects of hyperatomarin, a prenylated phloroglucinol from Hypericum annulatum Moris subsp. annulatum, in a panel of malignant cell lines. Phytomedicine. 2008;15(11):1010-5. https://doi.org/10.1016/j.phymed.2008.04.008PMid:18539018 Schempp CM, Kirkin V, Simon-Haarhaus B, Kersten A, Kiss J, Termeer CC B et al. Inhibition of tumour cell growth by hyperforin, a novel anticancer drug from St. John's wort that acts by induction of apoptosis. Oncogene. 2002;21(8):1242. https://doi.org/10.1038/sj.onc.1205190PMid:11850844 Song S, Xiong C, Zhou M, Lu W, Huang Q, Ku G et al. Small-animal PET of tumor damage induced by photothermal ablation with 64Cu-bis-DOTA-hypericin. J Nucl Med. 2011;52(5):792-9. https://doi.org/10.2967/jnumed.110.086116PMid:21498539 Mirmalek SA, Azizi MA, Jangholi E, Yadollah-Damavandi S, Javidi MA, Parsa Y et al. Cytotoxic and apoptogenic effect of hypericin, the bioactive component of Hypericum perforatum on the MCF-7 human breast cancer cell line. Cancer Cell Int. 2015;16(1):1-9. https://doi.org/10.1186/s12935-016-0279-4PMid:26865836 PMCid:PMC4748624 Hamilton HB, Hinton DR, Law RE, Gopalakrishna R, Su YZ, Chen ZH et al. Inhibition of cellular growth and induction of apoptosis in pituitary adenoma cell lines by the protein kinase C inhibitor hypericin: potential therapeutic application. J Neurosurg. 1996;85(2):329-34. https://doi.org/10.3171/jns.1996.85.2.0329PMid:8755764 Kim JI, Park JH, Park HJ, Choi SK, Lee KT. Induction of differentiation of the human histocytic lymphoma cell line U-937 by hypericin. Arch Pharm Res. 1998;21(1):41-5. https://doi.org/10.1007/BF03216751PMid:9875513 Acar M, Ocak Z, Erdogan K, Cetin EN, Hatipoglu OF, Uyeturk U et al. The effects of hypericin on ADAMTS and p53 gene expression in MCF-7 breast cancer cells. Cancer. 2014;3:5. Jendželovský R, MikeÅ¡ J, SouÄek K, Procházková J, Kello M, SaÄková V et al. Drug efflux transporters, MRP1 and BCRP, affect the outcome of hypericin-mediated photodynamic therapy in HT-29 adenocarcinoma cells. Photochem Photobiol Sci. 2009;8(12):1716-23. https://doi.org/10.1039/b9pp00086kPMid:20024169 Eriksson ES, Eriksson LA. Identifying the sarco (endo) plasmic reticulum Ca 2+ ATPase (SERCA) as a potential target for hypericin-a theoretical study. Phys Chem Chem Phys. 2012;14(36):12637-46. https://doi.org/10.1039/c2cp42237aPMid:22892582 Barliya T, Mandel M, Livnat T, Weinberger D, Lavie G. Degradation of HIF-1alpha under hypoxia combined with induction of Hsp90 polyubiquitination in cancer cells by hypericin: a unique cancer therapy. PloS One. 2011;6(9):e22849. https://doi.org/10.1371/journal.pone.0022849PMid:21949677 PMCid:PMC3176203 Peebles KA, Baker RK, Kurz EU, Schneider BJ, Kroll DJ. Catalytic inhibition of human DNA topoisomerase IIα by hypericin, a naphthodianthrone from St. John's wort (Hypericum perforatum). Biochem Pharmacol. 2001;62(8):1059-70. https://doi.org/10.1016/S0006-2952(01)00759-6
