Scutellarein treats neuroblastoma by regulating the expression of multiple targets

Abstract

The aim of this study is to investigate the effect of scutellarein on the proliferation of neuroblastoma cells and the underlying mechanism. Six cell lines were used with drug intervention. Cell Counting Kit-8 was used to select the best, namely, SH-SY5Y, and then its IC₅₀ value was determined. To further investigate the mechanism of scutellarin affecting SH-SY5Y proliferation, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of 11 factors. Scutellarin administration with 300 µM significantly reduced the number of SH-SY5Y, especially on the 3rd day of exposure to scutellarin. The IC₅₀ value of scutellarin in SH-SY5Y cells was determined to be 117.8 µM. But the practical results showed that 300 µM was the optimal concentration of scutellarin. qRT-PCR further detected upregulated maternally expressed gene 3 (MEG3), oncogene c-Fos (c-FOS), and c-jun and downregulated M2 isoform of pyruvate kinase (PKM2), non-SMC Condensin I Complex Subunit H (NCAPH), epidermal growth factor receptor (EGFR), transforming growth factor (TGF)-β1, and TGF-α, suggesting that scutellarin with 300 µM volume inhibited the survival of SH-SY5Y by regulating the expression of these 8 factors. Scutellarin could be a novel drug for the treatment of neuroblastoma, and its underlying mechanism may be related to the upregulated levels of MEG3, c-FOS, and c-jun and downregulated the expression of PKM2, NCAPH, EGFR, TGF-β1, and TGF-α.

Keywords

HS-SY5Y / neuroblastoma / proliferation inhibition / scutellarin

References

[1] ChungC, Boterberg T, LucasJ, et al. Neuroblastoma. Pediatr Blood Cancer. 2021;68(suppl 2):28473.

[2] ZafarA, WangW, LiuG, et al. Molecular targeting therapies for neuroblastoma: progress and challenges. Med Res Rev. 2021;41(2):961-1021.

[3] Young Jr.JL, RiesLG, SilverbergE, Horm JW, MillerRW. Cancer incidence, survival, and mortality for children younger than age 15 years. Cancer. 1986;58(suppl 2):598-602.

[4] LundbergKI, TreisD, JohnsenJI. Neuroblastoma heterogeneity, plasticity, and emerging therapies. Curr Oncol Rep. 2022;24(8):1053-1062.

[5] LouisCU, ShohetJM. Neuroblastoma: molecular pathogenesis and therapy. Annu Rev Med. 2015;66:49-63.

[6] MonclairT, Brodeur GM, AmbrosPF, et al. The International Neuroblastoma Risk Group (INRG) staging system: an INRG Task Force report. J Clin Oncol. 2009;27(2):298-303.

[7] Vénat-BouvetL, Le Brun-Ly V, MartinJ, GasnierO, Falkowsky S, Tubiana-MathieuN. Long-term survival in adult neuroblastoma with multiple recurrences. Case Rep Oncol. 2010;3(1):45-48.

[8] MarisJM, Hogarty MD, BagatellR, CohnSL. Neuroblastoma. Lancet. 2007;369(9579):2106-2120.

[9] JiangM, StankeJ, LahtiJM. The connections between neural crest development and neuroblastoma. Curr Top Dev Biol. 2011;94:77-127.

[10] LouXY, ChengJL, ZhangB. Therapeutic effect and mechanism of breviscapine on cisplatin-induced nephrotoxicity in mice. Asian Pac J Trop Med. 2015;8(10):873-877.

[11] ZhangHF, HuXM, WangLX, Xu SQ, ZengFD. Protective effects of scutellarin against cerebral ischemia in rats: evidence for inhibition of the apoptosis-inducing factor pathway. Planta Med. 2009;75(2):121-126.

[12] WangL, MaQ. Clinical benefits and pharmacology of scutellarin: a comprehensive review. Pharmacol Ther. 2018;190:105-127.

[13] LinLL, LiuAJ, LiuJG, Yu XH, QinLP, SuDF. Protective effects of scutellarin and breviscapine on brain and heart ischemia in rats. J Cardiovasc Pharmacol. 2007;50(3):327-332.

[14] QianL, ShenM, TangH, et al. Synthesis and protective effect of scutellarein on focal cerebral ischemia/reperfusion in rats. Molecules. 2012;17(9):10667-10674.

[15] KeY, BaoT, WuX, et al. Scutellarin suppresses migration and invasion of human hepatocellular carcinoma by inhibiting the STAT3/Girdin/Akt activity. Biochem Biophys Res Commun. 2017;483(1):509-515.

[16] LiH, HuangD, GaoZ, ChenY, ZhangL, Zheng J. Scutellarin inhibits the growth and invasion of human tongue squamous carcinoma through the inhibition of matrix metalloproteinase-2 and -9 and αvβ6 integrin. Int J Oncol. 2013;42(5):1674-1681.

[17] ZhuPT, MaoM, LiuZG, Tao L, YanBC. Scutellarin suppresses human colorectal cancer metastasis and angiogenesis by targeting ephrinb2. Am J Transl Res. 2017;9(11):5094-5104.

[18] SunCY, ZhuY, LiXF, et al. Scutellarin increases cisplatin-induced apoptosis and autophagy to overcome cisplatin resistance in non-small cell lung cancer via ERK/p53 and c-met/AKT signaling pathways. Front Pharmacol. 2018;9:92.

[19] GaoC, ZhouY, JiangZ, et al. Cytotoxic and chemosensitization effects of Scutellarin from traditional Chinese herb Scutellaria altissima L. in human prostate cancer cells. Oncol Rep. 2017;38(3):1491-1499.

[20] MamadalievaNZ, Herrmann F, El-ReadiMZ, et al. Flavonoids in Scutellaria immaculata and S. ramosissima (Lamiaceae) and their biological activity. J Pharm Pharmacol. 2011;63(10):1346-1357.

[21] AslanE, AdemS. In vitro effects of some flavones on human pyruvate kinase isoenzyme M2. J Biochem Mol Toxicol. 2015;29(3):109-113.

[22] WuWH, ChenTY, LuRW, ChenST, ChangCC. Benzoxazinoids from Scoparia dulcis (sweet broomweed) with antiproliferative activity against the DU-145 human prostate cancer cell line. Phytochemistry. 2012;83:110-115.

[23] KovalevichJ, Langford D. Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology. Methods Mol Biol. 2013;1078:9-21.

[24] BiedlerJL, Roffler-Tarlov S, SchachnerM, FreedmanLS. Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones. Cancer Res. 1978;38(11 pt 1):3751-3757.

[25] XieHR, HuLS, LiGY. SH-SY5Y human neuroblastoma cell line: in vitro cell model of dopaminergic neurons in Parkinson’s disease. Chin Med J. 2010;123(8):1086-1092.

[26] PresgravesSP, AhmedT, BorwegeS, Joyce JN. Terminally differentiated SH-SY5Y cells provide a model system for studying neuroprotective effects of dopamine agonists. Neurotoxic Res. 2003;5(8):579-598.

[27] XiongLL, DuRL, XueLL, et al. Anti-colorectal cancer effects of scutellarin revealed by genomic and proteomic analysis. Chin Med. 2020;15:28.

[28] BiedlerJL, HelsonL, SpenglerBA. Morphology and growth, tumorigenicity, and cytogenetics of human neuroblastoma cells in continuous culture. Cancer Res. 1973;33(11):2643-2652.

[29] LanttoT, LaaksoI, DormanH, et al. Cellular stress and p53-associated apoptosis by Juniperus communis L. berry extract treatment in the human SH-SY5Y neuroblastoma cells. Int J Mol Sci. 2016;17(7):1113.

[30] XuP, HouL, JuC, et al. Isatin inhibits the proliferation and invasion of SH-SY5Y neuroblastoma cells. Mol Med Rep. 2016;13(3):2757-2762.

[31] ZengY, ZhangX, HuangZ, et al. Intracellular Tat of human immunodeficiency virus type 1 activates lytic cycle replication of Kaposi’s sarcoma-associated herpesvirus: role of JAK/STAT signaling. J Virol. 2007;81(5):2401-2417.

[32] TsiambasE, Mastronikolis N, P FotiadesP, et al. c-Jun/c-Fos complex in laryngeal squamous cell carcinoma. J BUON. 2020;25(2):618-620.

[33] Milde-LangoschK. The Fos family of transcription factors and their role in tumourigenesis. Eur J Cancer. 2005;41(16):2449-2461.

[34] ZhangX, ZhouY, MehtaKR, et al. A pituitary-derived MEG3 isoform functions as a growth suppressor in tumor cells. J Clin Endocrinol Metab. 2003;88(11):5119-5126.

[35] BalikV, Srovnal J, SullaI, et al. MEG3: a novel long noncoding potentially tumour-suppressing RNA in meningiomas. J Neurooncol. 2013;112(1):1-8.

[36] LiH, LiB, ZhuD, et al. Downregulation of lncRNA MEG3 and miR-770-5p inhibit cell migration and proliferation in Hirschsprung’s disease. Oncotarget. 2017;8(41):69722-69730.

[37] ZhengY, LinL, ZhengZ. TGF-αinduces upregulation and nuclear translocation of Hes1 in glioma cell. Cell Biochem Funct. 2008;26(6):692-700.

[38] El-ObeidA, Hesselager G, WestermarkB, NistérM. TGF-α-driven tumor growth is inhibited by an EGF receptor tyrosine kinase inhibitor. Biochem Biophys Res Commun. 2002;290(1):349-358.

[39] AmmounS, Cunliffe CH, AllenJC, et al. ErbB/HER receptor activation and preclinical efficacy of lapatinib in vestibular schwannoma. Neuro Oncol. 2010;12(8):834-843.

[40] Vander HeidenMG, Cantley LC, ThompsonCB. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science. 2009;324(5930):1029-1033.

[41] MazurekS. Pyruvate kinase type M2: a key regulator of the metabolic budget system in tumor cells. Int J Biochem Cell Biol. 2011;43(7):969-980.

[42] MazurekS, Boschek CB, HugoF, EigenbrodtE. Pyruvate kinase type M2 and its role in tumor growth and spreading. Sem Cancer Biol. 2005;15(4):300-308.

[43] BluemleinK, Grüning NM, FeichtingerRG, LehrachH, KoflerB, RalserM. No evidence for a shift in pyruvate kinase PKM1 to PKM2 expression during tumorigenesis. Oncotarget. 2011;2(5):393-400.

[44] StetákA, VeressR, OvádiJ, CsermelyP, Kéri G, UllrichA. Nuclear translocation of the tumor marker pyruvate kinase M2 induces programmed cell death. Cancer Res. 2007;67(4):1602-1608.

[45] IqbalMA, GuptaV, GopinathP, Mazurek S, BamezaiRNK. Pyruvate kinase M2 and cancer: an updated assessment. FEBS Lett. 2014;588(16):2685-2692.

[46] WuQ, Kawahara M, KonoT. Synergistic role of Igf2 and Dlk1 in fetal liver development and hematopoiesis in bi-maternal mice. J Reprod Dev. 2008;54(3):177-182.

[47] KrauseTM. BCORL1 Regulates Stemness in Hepatoblastoma Cells Through Inhibition of KRT19 Expression. lmu;2020.

[48] SunY, WangX, WenH, ZhuB, YuL. Expression and clinical significance of the NCAPH, AGGF1, and FOXC2 proteins in serous ovarian cancer. Cancer Manag Res. 2021;13:7253-7262.

[49] OguraT, AzumaK, SatoJ, et al. OCT1 is a poor prognostic factor for breast cancer patients and promotes cell proliferation via inducing NCAPH. Int J Mol Sci. 2021;22(21):11505.

[50] LiB, XiaoQ, ShanL, Song Y. NCAPH promotes cell proliferation and inhibits cell apoptosis of bladder cancer cells through MEK/ERK signaling pathway. Cell Cycle. 2022;21(4):427-438.

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2023 The Authors. Ibrain published by Affiliated Hospital of Zunyi Medical University (AHZMU) and Wiley-VCH GmbH.