ISSN 0564-3783  

Main page
Information to authors
Editorial board
Mobile version

In Ukrainian

Export citations

4-Hydroxycoumarin effects on both cellular and genetic characteristics of hepatocellular carcinoma cells

Öner Ç., Soyergin D., Özyurt A., Çolak E.


4-Hydroxycoumarin is an aromatic substance which is metabolized in liver and used as a therapeutic agent for various diseases. We aimed to determine the impact of 4-Hydroxycoumarin on HepG2 cells according to their viability, proliferation, adhesion and gene expression of cellular behavior parameters. Inhibitory concentration 50 (IC50) of 4-Hydroxycoumarin was detected on HepG2 cells. After determining the optimal time and concentration, the effect of 4-Hydroxycoumarin on viability, proliferation and adhesion of HepG2 cells were observed. Gene expressions of Ki-67, MMP-2, MMP-9 and piR-823 expression were determined by using Real Time-Polymerase Chain Reaction. IC50 value of 4-Hydroxycoumarin on HepG2 cells was 5 M at the 48th hour (p < 0.001). 5 M at the 48th hour of 4-Hydroxycoumarin caused to decrease of proliferation (p < 0.001) and viability of HepG2 cells (p < 0.001). Viability rate were supported by hematoxylin-eosin staining. Adhesion of cells increased on 4-Hydroxycoumarin treated cells compared to control (p < 0.001). While Ki-67 gene expression of 4-Hydroxycoumarin treated group decreased (p < 0.001); upregulation of MMP-2, MMP-9 and piR-823 expressions were determined in 4-Hydroxycoumarin treated group (p < 0.001). According to the cellular and genetic perspective, 4-Hydroxycoumarin might be useful to treat hepatocellular carcinoma. High adhesion and proliferation are the main characteristics of HepG2 cells, 4-Hydroxycoumarin treatment caused to lose these functions. The genetic markers of these characteristics also supported the same result. These are first findings about the effect of 4-Hydroxycoumarin on piR-823 and genes which are key features of cellular survival mechanisms.

Key words: 4-Hydroxycoumarin, Matrix metalloproteinase, piRNA, piR-823, Hepatocellular carcinoma

Tsitologiya i Genetika 2022, vol. 56, no. 3, pp. 76-79

  1. Maltepe University, Medical Faculty, Department of Medical Biology and Genetics, İstanbul, Turkey
  2. Maltepe University, Medical Faculty, İstanbul, Turkey
  3. Eskişehir Osmangazi University, Medical Faculty, Department of Biostatistics, Eskişehir, Turkey

E-mail: cagri.oner

Öner Ç., Soyergin D., Özyurt A., Çolak E. 4-Hydroxycoumarin effects on both cellular and genetic characteristics of hepatocellular carcinoma cells, Tsitol Genet., 2022, vol. 56, no. 3, pp. 76-79.

In "Cytology and Genetics":
Çağri Öner, Dilara Soyergin, Ahmet Özyurt & Ertuğrul Çolak 4-Hydroxycoumarin Effects on Both Cellular and Genetic Characteristics of Hepatocellular Carcinoma Cells, Cytol Genet., 2022, vol. 56, no. 3, pp. 292300
DOI: 10.3103/S0095452722030094


Abraham, K., Wohrlin, F., Lindtner, O., et al., Toxicology and risk assessment of coumarin: Focus on human data, Mol. Nutr. Food Res., 2010, vol. 54, no. 2, pp. 228239.

Bar-Ziv, R., Tlusty, T., Moses, E., et al., Pearling in cells: A clue to understanding cell shape, Proc. Natl. Acad. Sci. U. S. A., 1999, vol. 96, no. 18, pp. 1014010145.

Breckenridge, A.M., Leck, J.B., Park, B.K., et al., J. Pharmacol., 1978, vol. 64, art. ID 399.

Bullwinkel, J., Baron-Luhr, B., Ludemann, A., et al., Ki-67 protein is associated with ribosomal RNA transcription in quiescent and proliferating cells, J. Cell. Physiol., 2006, vol. 206, no. 3, pp. 624635.

Chalbatani, G.M., Dana, H., Memari, F., et al., Biological function and molecular mechanism of piRNA in cancer, Pract. Lab. Med., 2019, vol. 13, art. ID e00113.

Chen, W.C., Liu, L., Shen, Y.F., et al., A new coumarin derivative plays a role in rhabdoviral clearance by interfering glycoprotein function during the early stage of viral infection, Cell. Signaling, 2018, vol. 51, pp. 199210.

Cheng, J., Deng, H.X., Xiao, B.X., et al., piR-823, a novel non-coding small RNA, demonstrates in vitro and in vivo tumor suppressive activity in human gastric cancer cells, Cancer Lett., 2012, vol. 315, pp. 1217.

Chien, Y.C., Liu, L.C., Ye, H.Y., et al., EZH2 promotes migration and invasion of triple-negative breast cancer cells via regulating TIMP2-MMP-2/-9 pathway, Am. J. Cancer Res., 2018, vol. 8, no. 3, pp. 422434.

Cui, J., Dong, B.W., Liang, P., et al., Effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma patients undergoing tumor resection, World J. Gastroenterol., 2004, vol. 10, pp. 15331536.

Cui, L., Lou, Y., Zhang, X., et al., Detection of circulating tumor cells in peripheral blood from patients with gastric cancer using piRNAs as markers, Clin. Biochem., 2011, vol. 44, no. 13, pp. 10501057.

Dehn, P.F., White, C.M., Conners, D.E., et al., Characterization of the human hepatocellular carcinoma (hepg2) cell line as an in vitro model for cadmium toxicity studies, In Vitro Cell. Dev. Biol.: Anim., 2004, vol. 40, nos. 56, pp. 172182.<172:COTHHC>2.0.CO;2

Deryugina, E.I. and Quigley, J.P., Matrix metalloproteinases and tumor metastasis, Cancer Metastasis Rev., 2006, vol. 25, pp. 934.

Dowsett, M., Nielsen, T.O., Ahern, R., et al., Assessment of Ki67 in breast cancer: Updated recommendations from the International Ki67 in Breast Cancer working group, J. Natl. Cancer Inst., 2011, vol. 103, no. 7, pp. 16561664.

Egan, D., James, P., Cooke, D., et al., Studies on the cytostatic and cytotoxic effects and mode of action of 8-nitro-7-hydroxycoumarin, Cancer Lett., 1997, vol. 118, no. 2, pp. 201211.

El-Serag, H.B. and Rudolph, K.L., Hepatocellular carcinoma: epidemiology and molecular carcinogenesis, Gastroenterology, 2007, vol. 132, no. 7, pp. 25572576.

Emami, S. and Dadashpour, S., Current developments of coumarin-based anti-cancer agents in medicinal chemistry, Eur. J. Med. Chem., 2015, vol. 102, pp. 611630.

Finn, G.J., Creaven, B., and Egan, D.A., Study of the in vitro cytotoxic potential of natural and synthetic coumarin derivatives using human normal and neoplastic skin cell lines, Melanoma Res., 2001, vol. 11, pp. 461467.

Funaya, C., Antony, C., Sachidanandam, R., et al., Miwi catalysis is required for piRNA amplification‑independent LINE1 transposon silencing, Nature, 2011, vol. 480, pp. 264267.

Gu, J., Li, Y., Fan, L., et al., Identification of aberrantly expressed long non-coding RNAs in stomach adenocarcinoma, Oncotarget, 2017, vol. 8, pp. 4920149216.

Han, H., Zhang, Z.-F., Zhang, J.-F., et al., Novel coumarin derivatives: Synthesis, anti-breast cancer activity and docking study, Main Group Chem., 2019, vol. 18, pp. 7179.

Iliev, R., Stanik, M., Fedorko, M., et al., Decreased expression levels of PIWIL1, PIWIL2, and PIWIL4 are associated with worse survival in renal cell carcinoma patients, OncoTargets Ther., 2016, vol. 9, pp. 217222.

Ingber, D.E., Tensegrity II. How structural networks influence cellular information processing networks, J. Cell Sci., 2003, vol. 116, no. 8, pp. 13971408.

Jacobs, D.I., Qin, Q., Lerro, M.C., et al., PIWI-interacting RNAs in gliomagenesis: evidence from post-GWAS and functional analyses, Cancer Epidemiol., Biomarkers Prev., 2016, vol. 25, no. 7, pp. 10731080.

Jimenez-Orozco, F.A., Lopez-Gonzalez, J.S., Nieto-Rodriguez, A., et al., Decrease of cyclin D1 in the human lung adenocarcinoma cell line A-427 by 7-hydroxycoumarin, Lung Cancer, 2001, vol. 34, no. 2, pp. 185194.

Zanker, K.S., Blumel, J., Lange, J.R., et al., Coumarin in melanoma patients: an experimental and clinical study, Drugs Exp. Clin. Res., 1984, vol. 10, pp. 767774.

Kawaii, S., Tomono, Y., Ogawa, K., et al., The antiproliferative effect of coumarins on several cancer cell lines, Anticancer Res., 2001, vol. 21, pp. 917923.

Keating, J.G. and Okennedy, R., The Chemistry and Occurrence of Coumarins, New York: Wiley, 1997.

King, K.L., Hwang, J.J., Chau, G.Y., et al., Ki-67 expression as a prognostic marker in patients with hepatocellular carcinoma, J. Gastroenterol. Hepatol., 1998, vol. 13, pp. 273279.

Kokron, O., Maca, S., Gasser, G., et al., Cimetidine and coumarin therapy of renal-cell carcinoma, Oncology, 1991, vol. 48, pp. 102126.

Lake, B.G., Coumarin metabolism, toxicity and carcinogenicity: relevance for human risk assessment, Food Chem. Toxicol., 1999, vol. 37, pp. 423453.

Li, J., Xue, X.Y., Li, X., et al., Synthesis of biscoumarin and dihydropyran derivatives as two novel classes of potential anti-bacterial derivatives, Arch. Pharmacal Res., 2016, vol. 39, pp. 13491355.

Lirdprapamongkol, K., Kramb, J.P., Suthiphongchai, T., et al., Vanillin suppresses metastatic potential of human cancer cells through PI3K inhibition and decreases angiogenesis in vivo, J. Agric. Food Chem., 2009, vol. 57, pp. 30553063.

Liu, Y., Dou, M., Song, X., et al., The emerging role of the piRNA/piwi complex in cancer, Mol. Cancer, 2019, vol. 18, art. ID 123.

Lopez-Gonzalez, J.S., Prado-Garcia, H., Aguilar-Cazares, D., et al., Apoptosis and cell cycle disturbances induced by coumarin and 7-hydroxycoumarin on human lung carcinoma cell lines, Lung Cancer, 2004, vol. 43, pp. 275283.

Man, S.L., Gao, W.Y., Zhang, Y.J., et al., Formosanin C‑inhibited pulmonary metastasis through repression of matrix metalloproteinases on mouse lung adenocarcinoma, Cancer Biol. Ther., 2011, vol. 11, pp. 592598.

Marshall, M.E., Butler, K., and Fried, A., Phase I evaluation of coumarin (1,2-benzopyrone) and cimetidine in patients with advanced malignancies, Mol. Biother., 1991, vol. 3, no. 3, pp. 170178.

Marshall, M.E., Kervin, K., Benefield, C., et al., Growth-inhibitory effects of coumarin (1,2-benzopyrone) and 7-hydroxycoumarin on human malignant cell lines in vitro, J. Cancer Res. Clin. Oncol., 1994, vol. 120, pp. S3S10.

Marusyk, A., Tabassum, D.P., Altrock, P.M., et al., Non-cell-autonomous driving of tumour growth supports sub-clonal heterogeneity, Nature, 2014, vol. 514, pp. 5458.

Mohler, J.L., Gomella, L.G., Crawford, E.D., et al., Phase II evaluation of coumarin (1,2-benzopyrone) in metastatic prostatic carcinoma, Prostate, 1992, vol. 20, no. 2, pp. 123131.

Mohler, J.L., Williams, B.T., Thompson, I.M., et al., Coumarin (1,2-benzopyrone) for the treatment of prostatic carcinoma, J. Cancer Res. Clin. Oncol., 1994, vol. 120, pp. S35S38.

Mosmann, T., Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays, J. Immunol. Methods, 1983, vol. 65, nos. 12, pp. 5563.

Nasr, T., Bondock, S., and Youns, M., Anticancer activity of new coumarin substituted hydrazide-hydrazone derivatives, Eur. J. Med. Chem., 2014, vol. 76, pp. 539548.

Öner, C., Turgut Coşan, D., and Çolak, E., Estrogen and androgen hormone levels modulate the expression of PIWI interacting RNA in prostate and breast cancer, PLoS One, 2016, vol. 11, art. ID e0159044.

Ozata, D.M., Gainetdinov, I., Zoch, A., et al., PIWI-interacting RNAs: small RNAs with big functions, Nat. Rev. Genet., 2019, vol. 20, pp. 89108.

Phillips, C., Zeringue, A.L., Mcdonald, J.R., et al., Tumor necrosis factor inhibition and head and neck cancer recurrence and death in rheumatoid arthritis, PLoS One, 2015, vol. 10, art. ID e0143286.

Rennenberg, R.J., Van Varik, B.J., Schurgers, L.J., et al., Chronic coumarin treatment is associated with increased extracoronary arterial calcification in humans, Blood, 2010, vol. 115, no. 24, pp. 51215123.

Salinas-Jazmin, N., De La Fuente, M., Jaimez, R., et al., Antimetastatic, antineoplastic, and toxic effects of 4-hydroxycoumarin in a preclinical mouse melanoma model, Cancer Chemother. Pharmacol., 2010, vol. 65, pp. 931940.

Shokoohinia, Y., Hosseinzadeh, L., Alipour, M., et al., Comparative evaluation of cytotoxic and apoptogenic effects of several coumarins on human cancer cell lines: Osthole induces apoptosis in p53-deficient H1299 cells, Adv. Pharmacol. Sci. 2014, art. ID 847574.

Sobecki, M., Mrouj, K., Colinge, J., et al., Cell-Cycle regulation accounts for variability in Ki-67 expression levels, Cancer Res., 2017, vol. 77, no. 10, pp. 27222734.

Stefanova, T.H., Nikolova, N.J., Toshkova, R.A., et al., Antitumor and immunomodulatory effect of coumarin and 7-hydroxycoumarin against Sarcoma 180 in mice, J. Exp. Ther. Oncol., 2007, vol. 6, no. 2, pp. 107115.

Thornes, R.D., Daly, L., Lynch, G., et al., Treatment with coumarin to prevent or delay recurrence of malignant melanoma, J. Cancer Res. Clin. Oncol., 1994, vol. 120, pp. S32S34.

Ujjin, P., Satarug, S., Vanavanitkun, Y., et al., Variation in coumarin 7-hydroxylase activity associated with genetic polymorphism of cytochrome P450 2A6 and the body status of iron stores in adult Thai males and females, Pharmacogenetics, 2002, vol. 12, no. 3 pp. 241249.

Vázquez, R., Riveiro, M.E., Vermeulen, M., et al., Structure-anti-leukemic activity relationship study of ortho-dihydroxycoumarins in U-937 cells: key role of the δ-lactone ring in determining differentiation-inducing potency and selective pro-apoptotic action, Bioorg. Med. Chem., 2012, vol. 20, pp. 55375549.

Velasco-Velazquez, M.A., Agramonte-Hevia, J., Barrera, D., et al., 4-Hydroxycoumarin disorganizes the actin cytoskeleton in B16F10 melanoma cells but not in B82 fibroblasts, decreasing their adhesion to extracellular matrix proteins and motility, Cancer Lett., 2003, vol. 198, pp. 179186.

Velasco-Velázquez, M.A., Salinas-Jazmín, N., Mendoza-Patiño, N., et al., Reduced paxillin expression contributes to the antimetastatic effect of 4-hydroxycoumarin on B16-F10 melanoma cells, Cancer Cell. Int., 2008, art. ID 8.

Vianna, D.R., Hamerski, L., Figueiro, F., et al., Selective cytotoxicity and apoptosis induction in glioma cell lines by 5-oxygenated-6,7-methylenedioxycoumarins from Pterocaulon species, Eur. J. Med. Chem., 2012, vol. 57, pp. 268274.

Wu, X.Q., Huang, C., Jia, Y.M., et al., Novel coumarin-dihydropyrazole thio-ethanone derivatives: Design, synthesis and anticancer activity, Eur. J. Med. Chem., 2014, vol. 74, pp. 717725.

Xiao, Z., Shen, J., Zhang, L., et al., Therapeutic targeting of noncoding RNAs in hepatocellular carcinoma: Recent progress and future prospects (Review), Oncol. Lett., 2018, vol. 15, pp. 33953402.

Xu, C., Rao, Y.S., Xu, B., et al., An in vivo pilot study characterizing the new CYP2A6*7, *8, and *10 alleles, Biochem Biophys. Res. Commun., 2002, vol. 290, no. 1, pp. 318324.

Yamashiro, H. and Siomi, M.C., PIWI-interacting RNA in Drosophila: Biogenesis, transposon regulation, and beyond., Chem. Rev., 2018, vol. 118, no. 8, pp. 44044421.

Yan, H., Wu, Q.L., Sun, C.Y., et al., piRNA-823 contributes to tumorigenesis by regulating de novo DNA methylation and angiogenesis in multiple myeloma, Leukemia, 2015, vol. 29, pp. 196206.

Yang, Y., Lu, N., Zhou, J., et al., Cyclophilin A up-regulates MMP-9 expression and adhesion of monocytes/macrophages via CD147 signalling pathway in rheumatoid arthritis, Rheumatology (Oxford), 2008, vol. 47, no. 9, pp. 12991310.

Yin, J., Jiang, X.Y., Qi, W., et al., piR-823 contributes to colorectal tumorigenesis by enhancing the transcriptional activity of HSF1, Cancer Sci., 2017, vol. 108, no. 9, pp. 17461756.

Zhang, J., Zhang, D., Wu, G-Q., et al., Propofol inhibits the adhesion of hepatocellular carcinoma cells by upregulating microRNA-199a and downregulating MMP-9 expression, Hepatobiliary Pancreatic Dis. Int., 2013, vol. 12, pp. 305309.

Copyright© ICBGE 2002-2023 Coded & Designed by Volodymyr Duplij Modified 01.02.23