TY - JOUR T1 - Myricetin can control metastasis and invasion by suppressing ATF2-related signaling pathway in Rapamycin-resistant HepG2 hepatocellular cancer cells AU - Erdoğan, Ömer AU - Çevik, Özge PY - 2025 DA - June JF - Journal of Research in Pharmacy JO - J. Res. Pharm. PB - Marmara University WT - DergiPark SN - 2630-6344 SP - 557 EP - 565 VL - 27 IS - 2 LA - en AB - Hepatocellular carcinoma continues to be one of the major health problems. One of the most importantchallenges in the treatment of hepatocellular cancer is gaining resistance to chemotherapy. Therefore, there is an urgentneed for discoveries of naturally occurring antitumor compounds in the diet and to elucidate the anticancer mechanismsof action of existing dietary origin compounds. Myricetin is a dietary flavonoid and its antitumor activity is reported.In this study, HepG2 cells were incubated with 100 nM rapamycin for 6 weeks to develop rapamycin resistance. 10, 100,250 and 500 µM myricetin was applied to rapamycin resistant HepG2 human hepatocellular cancer cells (HepG2-RR)for 24 and 48 hours. Changes in ATF2, SMAD3, SMAD4 and Col1A1 expressions of HepG2-RR cells after myricetinapplication were evaluated by Western blot and qPCR analysis. In addition, the effect of myricetin treatment on themigration/invasion of HepG2-RR cells was specified using the BD 24-well Boyden chamber. The IC50 values ofrapamycin and myricetin on HepG2 cells for 48 hours incubation was calculated as 89,03±6,14 nM and 126,25±7,32 µM,respectively. On the other hand, we found that HepG2-RR cells treated with myricetin decreased cell proliferation,invasion, and migration by reducing the expression of ATF2, SMAD3, SMAD4, and Col1A1. As a result, we were shownthat myricetin administration inhibited cell growth and cell migration by suppressing ATF2 in hepatocellular carcinomacells that gained drug resistance. KW - Myricetin KW - Rapamycin KW - Liver Cancer KW - Metastasis CR - Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians. 2021; 71(3): 209-249. https://doi.org/10.3322/caac.21660 CR - Liu CY, Chen KF, Chen PJ. Treatment of liver cancer. Cold Spring Harbor perspectives in medicine. 2015; (9): a021535. Anwanwan D, Singh, SK, Singh S, Saikam V, Singh R. Challenges in liver cancer and possible treatment approaches. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer. 2020; 1873(1): 188314. https://doi.org/10.1016/j.bbcan.2019.188314 CR - Bhoumik A, Lopez‐Bergami P, Ronai ZE. ATF2 on the double–activating transcription factor and DNA damage response protein. Pigment Cell Research. 2007; 20(6): 498-506. https://doi.org/10.1111/j.1600-0749.2007.00414.x CR - Bao ZM, Yao D, Qian X, Zhang HG, Yang M, Guo YH, Qin L. (2022). Activating transcription factor 2 promotes the progression of hepatocellular carcinoma by inducing the activation of the WHSC1‐mediated TOP2A/PI3K/AKT axis. The Kaohsiung Journal of Medical Sciences. 2022; 1-13. https://doi.org/10.1002/kjm2.12536 CR - Luo L, Cai L, Luo L, Tang Z, Meng X. Silencing activating transcription factor 2 promotes the anticancer activity of sorafenib in hepatocellular carcinoma cells. Molecular medicine reports. 2018; 17(6): 8053-8060. https://doi.org/10.3892/mmr.2018.8921 CR - Sarbassov DD, Ali SM, Sabatini DM. Growing roles for the mTOR pathway. Current opinion in cell biology. 2005; 17(6); 596-603. https://doi.org/10.1016/j.ceb.2005.09.009 CR - Sarbassov DD, Ali SM, Sengupta S, Sheen, JH, Hsu PP, Bagley AF, Sabatini DM. Prolonged rapamycin treatment inhibits mTORC2 assembly and Akt/PKB. Molecular cell. 2006; 22(2): 159-168. https://doi.org/10.1016/j.molcel.2006.03.029 CR - Guertin DA, Sabatini DM. An expanding role for mTOR in cancer. Trends Mol. Med. 2005; 11: 353–361. https://doi.org/10.1016/j.molmed.2005.06.007 CR - Polivka J, Janku F. Molecular targets for cancer therapy in the PI3K/AKT/mTOR pathway. Pharmacology & therapeutics. 2014: 142(2); 164-175. https://doi.org/10.1016/j.pharmthera.2013.12.004 CR - Devi KP, Rajavel T, Habtemariam S, Nabavi SF, Nabavi SM. Molecular mechanisms underlying anticancer effects of myricetin. Life sciences. 2015; 142: 19-25. https://doi.org/10.1016/j.lfs.2015.10.004 CR - Li C, Lim SC, Kim J, Choi JS. Effects of myricetin, an anticancer compound, on the bioavailability and pharmacokinetics of tamoxifen and its main metabolite, 4-hydroxytamoxifen, in rats. European journal of drug metabolism and pharmacokinetics. 2011: 36(3); 175-182. https://doi.org/10.1007/s13318-011-0036-y CR - Gorshtein A, Rubinfeld H, Kendler E, Theodoropoulou M, Cerovac V, Stalla GK, Shimon I. (2009). Mammalian target of rapamycin inhibitors rapamycin and RAD001 (everolimus) induce anti-proliferative effects in GH-secreting pituitary tumor cells in vitro. Endocrine-related cancer. 2009: 16(3); 1017-1027. https://doi.org/10.1677/ERC-08-0269 CR - Koike H, Nitta T, Sekine Y, Arai S, Furuya Y, Nomura M, Suzuki K. (2014). YM155 reverses rapamycin resistance in renal cancer by decreasing survivin. Journal of cancer research and clinical oncology. 2014: 140; 1705-1713. https://doi.org/10.1007/s00432-014-1734-z CR - Tian X, Dai S, Sun J, Jiang S, Sui C, Meng F, Jiang Y. Inhibition of MDM2 re-sensitizes rapamycin resistant renal cancer cells via the activation of p53. Cellular Physiology and Biochemistry. 2016: 39(5); 2088-2098. https://doi.org/10.1159/000447904 CR - Li QL, Gu FM, Wang Z, Jiang JH, Yao LQ, Tan CJ, Zhou J. Activation of PI3K/AKT and MAPK pathway through a PDGFRβ-dependent feedback loop is involved in rapamycin resistance in hepatocellular carcinoma. PLoS One. 2012: 7(3); e33379. https://doi.org/10.1371/journal.pone.0033379 CR - Zugazagoitia J, Guedes C, Ponce S, Ferrer I, Molina-Pinelo S, Paz-Ares L. Current challenges in cancer treatment. Clinical therapeutics. 2016; 38(7): 1551-1566. https://doi.org/10.1016/j.clinthera.2016.03.026 CR - Sun F, Zheng XY, Ye J, Wu TT, Wang JL, Chen W. Potential anticancer activity of myricetin in human T24 bladder cancer cells both in vitro and in vivo. Nutrition and cancer. 2012: 64(4); 599-606. https://doi.org/10.1080/01635581.2012.665564 CR - Neuzillet C, Tijeras-Raballand A, de Mestier L, Cros J, Faivre S, Raymond E. MEK in cancer and cancer therapy. Pharmacol. Ther. 2014: 141; 160–171. https://doi.org/10.1016/j.pharmthera.2013.10.001 CR - Lee KW, Kang NJ, Rogozin EA, Kim HG, Cho YY, Bode AM, Dong Z. Myricetin is a novel natural inhibitor of neoplastic cell transformation and MEK1. Carcinogenesis, 2007: 28(9); 1918-1927. https://doi.org/10.1093/carcin/bgm110 CR - Derynck R, Akhurst RJ, Balmain A. TGF-b signaling in tumor suppression and cancer progression. Nat. Genet. 2001; 29; 117–129. https://doi.org/10.1038/ng1001-117 CR - Tang PMK, Zhou S, Meng XM, Wang QM, Li CJ, Lian GY, Lan HY. Smad3 promotes cancer progression by inhibiting E4BP4-mediated NK cell development. Nature communications. 2017: 8(1); 1-15. https://doi.org/10.1038/ncomms14677 CR - Alhopuro P, Alazzouzi H, Sammalkorpi H, Dávalos V, Salovaara R, Hemminki A, Arango D. (2005). SMAD4 levels and response to 5-fluorouracil in colorectal cancer. Clinical Cancer Research. 2005: 11(17); 6311-6316. https://doi.org/10.1158/1078-0432.CCR-05-0244 CR - Li J, Ding Y, Li A. Identification of COL1A1 and COL1A2 as candidate prognostic factors in gastric cancer. World journal of surgical oncology. 2016: 14(1); 1-5. https://doi.org/10.1186/s12957-016-1056-5 CR - Liu S, Liao G, Li G. Regulatory effects of COL1A1 on apoptosis induced by radiation in cervical cancer cells. Cancer cell international. 2017: 17(1); 1-9. https://doi.org/10.1186/s12935-017-0443-5 CR - Zhao W, Jiang X, Yang, S. lncRNA TUG1 promotes cell proliferation, migration, and invasion in hepatocellular carcinoma via regulating miR-29c-3p/COL1A1 axis. Cancer Management and Research. 2020: 12; 6837. https://doi.org/10.2147/CMAR.S256624 CR - Gumus E, Erdogan O, Demirkan B, Cevik O. Squalene suppresses Jam-A, Claudin 5, and Occludin by accelerating cell death and reducing neuronal interaction in the neuroblastoma cell line SH-SY5Y. Journal of Research in Pharmacy. 2022: 26(6); 1676-1684. http://dx.doi.org/10.29228/jrp.258 CR - Erdogan O, Abbak M, Demirbolat GM, Birtekocak F, Aksel M, Pasa S, Cevik, O. (2019). Green synthesis of silver nanoparticles via Cynara scolymus leaf extracts: The characterization, anticancer potential with photodynamic therapy in MCF7 cells. PloS one. 2019: 14(6); e0216496. https://doi.org/10.1371/journal.pone.0216496 CR - Paşa S, Erdogan O, Cevik O. Design, synthesis and investigation of procaine based new Pd complexes as DNA methyltransferase inhibitor on gastric cancer cells. Inorganic Chemistry Communications. 2021: 132; 108846. https://doi.org/10.1016/j.inoche.2021.108846 CR - Cevik O, Li D, Baljinnyam E, Manvar D, Pimenta EM, Waris G, Kaushik-Basu, N. Interferon regulatory factor 5 (IRF5) suppresses hepatitis C virus (HCV) replication and HCV-associated hepatocellular carcinoma. Journal of Biological Chemistry. 2017: 292(52); 21676-21689. https://doi.org/10.1074/jbc.M117.792721 UR - https://dergipark.org.tr/en/pub/jrespharm/issue//1690367 L1 - https://dergipark.org.tr/en/download/article-file/4832632 ER -