Full Text:   <976>

Summary:  <939>

Suppl. Mater.: 

CLC number: 

On-line Access: 0000-00-00

Received: 2020-08-11

Revision Accepted: 2020-11-16

Crosschecked: 0000-00-00

Cited: 0

Clicked: 2005

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Ping XU

https://orcid.org/0000-0003-1599-7408

-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2021 Vol.22 No.7 P.548-562

http://doi.org/10.1631/jzus.B2000455


Potential effect of EGCG on the anti-tumor efficacy of metformin in melanoma cells


Author(s):  An’an XU, Jeehyun LEE, Yueling ZHAO, Yuefei WANG, Xiaoli LI, Ping XU

Affiliation(s):  Department of Tea Science, Zhejiang University, Hangzhou 310058, China; more

Corresponding email(s):   zdxp@zju.edu.cn

Key Words:  Epigallocatechin-3-gallate (EGCG), Metformin, Melanoma, Anti-tumor efficacy


An’an XU, Jeehyun LEE, Yueling ZHAO, Yuefei WANG, Xiaoli LI, Ping XU. Potential effect of EGCG on the anti-tumor efficacy of metformin in melanoma cells[J]. Journal of Zhejiang University Science B, 2021, 22(7): 548-562.

@article{title="Potential effect of EGCG on the anti-tumor efficacy of metformin in melanoma cells",
author="An’an XU, Jeehyun LEE, Yueling ZHAO, Yuefei WANG, Xiaoli LI, Ping XU",
journal="Journal of Zhejiang University Science B",
volume="22",
number="7",
pages="548-562",
year="2021",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2000455"
}

%0 Journal Article
%T Potential effect of EGCG on the anti-tumor efficacy of metformin in melanoma cells
%A An’an XU
%A Jeehyun LEE
%A Yueling ZHAO
%A Yuefei WANG
%A Xiaoli LI
%A Ping XU
%J Journal of Zhejiang University SCIENCE B
%V 22
%N 7
%P 548-562
%@ 1673-1581
%D 2021
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2000455

TY - JOUR
T1 - Potential effect of EGCG on the anti-tumor efficacy of metformin in melanoma cells
A1 - An’an XU
A1 - Jeehyun LEE
A1 - Yueling ZHAO
A1 - Yuefei WANG
A1 - Xiaoli LI
A1 - Ping XU
J0 - Journal of Zhejiang University Science B
VL - 22
IS - 7
SP - 548
EP - 562
%@ 1673-1581
Y1 - 2021
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2000455


Abstract: 
metformin, a first-line drug for type 2 diabetes mellitus, has been recognized as a potential anti-tumor agent in recent years. epigallocatechin-3-gallate (EGCG), as the dominant catechin in green tea, is another promising adjuvant agent for tumor prevention. In the present work, the potential effect of EGCG on the anti-tumor efficacy of metformin in a mouse melanoma cell line (B16F10) was investigated. Results indicated that EGCG and metformin exhibited a synergistic effect on cell viability, migration, and proliferation, as well as signal transducer and activator of transcription 3/nuclear factor-κB (STAT3/NF-κB) pathway signaling and the production of inflammation cytokines. Meanwhile, the combination showed an antagonistic effect on cell apoptosis and oxidative stress levels. The combination of EGCG and metformin also differentially affected the nucleus (synergism) and cytoplasm (antagonism) of B16F10 cells. Our findings provide new insight into the potential effects of EGCG on the anti-tumor efficacy of metformin in melanoma cells.

表没食子儿茶素-3-没食子酸酯(EGCG)对二甲双胍抗黑素瘤细胞的潜在影响

目的:探明表没食子儿茶素-3-没食子酸酯(EGCG)联合二甲双胍对高转移性小鼠黑素瘤细胞系B16F10活力、迁移和增殖等生长状态以及信号传导与转录活化因子3/核因子κB(STAT3/NF-κB)信号通路的作用,并利用拉曼光谱等分析EGCG和二甲双胍联合作用的潜在机制。
创新点:(1)EGCG联合二甲双胍对B16F10细胞活力、增殖以及STAT3/NF-κB信号通路等具有协同抑制作用;(2)EGCG联合二甲双胍在B16F10细胞凋亡和氧化应激方面具有拮抗作用;(3)利用拉曼光谱分析技术发现EGCG与二甲双胍联用对B16F10细胞的细胞核和细胞质存在差异化影响。
方法:利用CCK8测定细胞增殖,用细胞划痕实验测定细胞运动特性,利用流式细胞仪分析凋亡和细胞周期,利用免疫组织化学染色法观察细胞切片,用酶联免疫吸附试验(ELISA)测定炎症细胞因子和抗氧化酶的蛋白水平含量,用定量逆转录聚合酶链反应(qRT-PCR)检测其基因水平含量,用蛋白质印迹(western blot)探究其可能影响的细胞通路,用拉曼光谱技术从亚细胞水平探究作用机制。
结论:本研究表明EGCG与二甲双胍联合作用在抑制B16F10细胞生长、炎症水平和STAT3/NF-κB通路方面发挥部分协同效应。然而,这两种化合物对B16F10细胞凋亡和氧化应激具有拮抗作用。为了进一步证实EGCG和二甲双胍对黑色素瘤潜在的联合作用,还需要进一步研究EGCG和二甲双胍的药效学相互作用。

关键词:表没食子儿茶素-3-没食子酸酯(EGCG);二甲双胍;黑色素瘤;抗肿瘤功效

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]AhmadE, SargeantJA, ZaccardiF, et al., 2020. Where does metformin stand in modern day management of type 2 diabetes? Pharmaceuticals, 13(12):427.

[2]BalkwillF, MantovaniA, 2001. Inflammation and cancer: back to Virchow? Lancet, 357(9255):539-545.

[3]CaltagironeS, RossiC, PoggiA, et al., 2000. Flavonoids apigenin and quercetin inhibit melanoma growth and metastatic potential. Int J Cancer, 87(4):595-600.

[4]CerezoM, TichetM, AbbeP, et al., 2013. Metformin blocks melanoma invasion and metastasis development in AMPK/p53-dependent manner. Mol Cancer Ther, 12(8):1605-1615.

[5]ChaeYK, AryaA, MalecekMK, et al., 2016. Repurposing metformin for cancer treatment: current clinical studies. Oncotarget, 7(26):40767-40780.

[6]ChaiEZP, ShanmugamMK, ArfusoF, et al., 2016. Targeting transcription factor STAT3 for cancer prevention and therapy. Pharmacol Ther, 162:86-97.

[7]ChenK, QianWK, JiangZD, et al., 2017. Metformin suppresses cancer initiation and progression in genetic mouse models of pancreatic cancer. Mol Cancer, 16:131.

[8]ChenSN, ZhuXM, LaiXF, et al., 2014. Combined cancer therapy with non-conventional drugs: all roads lead to AMPK. Mini Rev Med Chem, 14(8):642-654.

[9]ChikaraS, NagaprashanthaLD, SinghalJ, et al., 2018. Oxidative stress and dietary phytochemicals: role in cancer chemoprevention and treatment. Cancer Lett, 413:122-134.

[10]CourtoisS, LehoursP, BessèdeE, 2019. The therapeutic potential of metformin in gastric cancer. Gastric Cancer, 22(4):653-662.

[11]de Souza NetoFP, BernardesSS, MarinelloPC, et al., 2017. Metformin: oxidative and proliferative parameters in-vitro and in-vivo models of murine melanoma. Melanoma Res, 27(6):536-544.

[12]EllisLZ, LiuWM, LuoYC, et al., 2011. Green tea polyphenol epigallocatechin-3-gallate suppresses melanoma growth by inhibiting inflammasome and IL-1β secretion. Biochem Biophys Res Commun, 414(3):551-556.

[13]FarhoodB, NajafiM, SalehiE, et al., 2019. Disruption of the redox balance with either oxidative or anti-oxidative overloading as a promising target for cancer therapy. J Cell Biochem, 120(1):71-76.

[14]FruehaufJP, MeyskensFL, 2007. Reactive oxygen species: a breath of life or death? Clin Cancer Res, 13(3):789-794.

[15]FujikiH, SueokaE, WatanabeT, et al., 2015. Primary cancer prevention by green tea, and tertiary cancer prevention by the combination of green tea catechins and anticancer compounds. J Cancer Prev, 20(1):1-4.

[16]HanahanD, WeinbergRA, 2011. Hallmarks of cancer: the next generation. Cell, 144(5):646-674.

[17]HartPC, ChiyodaT, LiuXJ, et al., 2019. SPHK1 is a novel target of metformin in ovarian cancer. Mol Cancer Res, 17(4):870-881.

[18]HartmanRI, LinJY, 2019. Cutaneous melanoma—a review in detection, staging, and management. Hematol Oncol Clin North Am, 33(1):25-38.

[19]HeathJR, RibasA, MischelPS, 2016. Single-cell analysis tools for drug discovery and development. Nat Rev Drug Discov, 15(3):204-216.

[20]HodgesV, TucciM, BenghuzziH, 2015. The effects of metformin and EGCG on PANC-1 cell survival. Biomed Sci Instrum, 51:393-399.

[21]Iglesias-PenaN, ParadelaS, Tejera-VaquerizoA, et al., 2019. Cutaneous melanoma in the elderly: review of a growing problem. Actas Dermosifiliogr, 110(6):434-447.

[22]JanjetovicK, Harhaji-TrajkovicL, Misirkic-MarjanovicM, et al., 2011. In vitro and in vivo anti-melanoma action of metformin. Eur J Pharmacol, 668(3):373-382.

[23]JauneE, RocchiS, 2018. Metformin: focus on melanoma. Front Endocrinol, 9:472.

[24]KarinM, 2006. Nuclear factor-κB in cancer development and progression. Nature, 441(7092):431-436.

[25]KimDS, ParkSH, KwonSB, et al., 2004. (-)-Epigallocatechin-3-gallate and hinokitiol reduce melanin synthesis via decreased MITF production. Arch Pharm Res, 27(3):334-339.

[26]KimK, YangWH, JungYS, et al., 2020. A new aspect of an old friend: the beneficial effect of metformin on anti-tumor immunity. BMB Rep, 53(10):512-520.

[27]KlaunigJE, 2018. Oxidative stress and cancer. Curr Pharm Des, 24(40):4771-4778.

[28]LeeJH, KishikawaM, KumazoeM, et al., 2010. Vitamin A enhances antitumor effect of a green tea polyphenol on melanoma by upregulating the polyphenol sensing molecule 67-kDa laminin receptor. PLoS ONE, 5(6):e11051.

[29]LiF, ZhangJW, ArfusoF, et al., 2015. NF-κB in cancer therapy. Arch Toxicol, 89(5):711-731.

[30]LiK, ZhangTT, WangF, et al., 2018. Metformin suppresses melanoma progression by inhibiting KAT5-mediated SMAD3 acetylation, transcriptional activity and TRIB3 expression. Oncogene, 37(22):2967-2981.

[31]LiuMH, ZhangZ, WangH, et al., 2019. Activation of AMPK by metformin promotes renal cancer cell proliferation under glucose deprivation through its interaction with PKM2. Int J Biol Sci, 15(3):617-627.

[32]LordSR, ChengWC, LiuD, et al., 2018. Integrated pharmacodynamic analysis identifies two metabolic adaption pathways to metformin in breast cancer. Cell Metab, 28(5):679-688.e4.

[33]LuCC, ChiangJH, TsaiFJ, et al., 2019. Metformin triggers the intrinsic apoptotic response in human AGS gastric adenocarcinoma cells by activating AMPK and suppressing mTOR/AKT signaling. Int J Oncol, 54(4):1271-1281.

[34]MaLW, WeiJW, WanJH, et al., 2019. Low glucose and metformin-induced apoptosis of human ovarian cancer cells is connected to ASK1 via mitochondrial and endoplasmic reticulum stress-associated pathways. J Exp Clin Cancer Res, 38(1):77.

[35]MignoletA, WoodBR, GoormaghtighE, 2018. Intracellular investigation on the differential effects of 4 polyphenols on MCF-7 breast cancer cells by Raman imaging. Analyst, 143:258-269.

[36]MillerKD, NogueiraL, MariottoAB, et al., 2019. Cancer treatment and survivorship statistics, 2019. CA Cancer J Clin, 69(5):363-385.

[37]MovasaghiZ, RehmanS, RehmanIU, 2007. Raman spectroscopy of biological tissues. Appl Spectrosc Rev, 42(5):493-541.

[38]NicolucciA, CharbonnelB, GomesMB, et al., 2019. Treatment patterns and associated factors in 14 668 people with type 2 diabetes initiating a second-line therapy: results from the global DISCOVER study programme. Diabetes, Obes Metab, 21(11):2474-2485.

[39]NiehrF, von EuwE, AttarN, et al., 2011. Combination therapy with vemurafenib (PLX4032/RG7204) and metformin in melanoma cell lines with distinct driver mutations. J Transl Med, 9:76.

[40]NihalM, AhsanH, SiddiquiIA, et al., 2009. (-)-Epigallocatechin-3-gallate (EGCG) sensitizes melanoma cells to interferon induced growth inhibition in a mouse model of human melanoma. Cell Cycle, 8(13):2057-2063.

[41]NihalM, RoelkeCT, WoodGS, 2010. Anti-melanoma effects of vorinostat in combination with polyphenolic antioxidant (-)-epigallocatechin-3-gallate (EGCG). Pharm Res, 27(6):1103-1114.

[42]PashaM, SivaramanSK, FrantzR, et al., 2019. Metformin induces different responses in clear cell renal cell carcinoma Caki cell lines. Biomolecules, 9(3):113.

[43]PerryG, RainaAK, NunomuraA, et al., 2000. How important is oxidative damage? Lessons from Alzheimer’s disease. Free Radical Bio Med, 28(5):831-834.

[44]RavindranathMH, RamasamyV, MoonS, et al., 2009. Differential growth suppression of human melanoma cells by tea (Camellia sinensis) epicatechins (ECG, EGC and EGCG). Evid Based Complement Alternat Med, 6(4):523-530.

[45]SabryD, AbdelaleemOO, el Amin AliAM, et al., 2019. Anti-proliferative and anti-apoptotic potential effects of epigallocatechin-3-gallate and/or metformin on hepatocellular carcinoma cells: in vitro study. Mol Biol Rep, 46(2):2039-2047.

[46]SchumacherB, GarinisGA, HoeijmakersJHJ, 2008. Age to survive: DNA damage and aging. Trends Genet, 24(2):77-85.

[47]ShenQ, TianF, JiangP, et al., 2009. EGCG enhances TRAIL-mediated apoptosis in human melanoma A375 cell line. J Huazhong Univ Sci Technol Med Sci, 29:771.

[48]SiegelRL, MillerKD, JemalA, 2019. Cancer statistics, 2019. CA Cancer J Clin, 69(1):7-34.

[49]SunRJ, ZhaiRR, MaCL, et al., 2020. Combination of aloin and metformin enhances the antitumor effect by inhibiting the growth and invasion and inducing apoptosis and autophagy in hepatocellular carcinoma through PI3K/AKT/mTOR pathway. Cancer Med, 9(3):1141-1151.

[50]SunY, HuangLQ, MackenzieGG, et al., 2011. Oxidative stress mediates through apoptosis the anticancer effect of phospho-nonsteroidal anti-inflammatory drugs: implications for the role of oxidative stress in the action of anticancer agents. J Pharmacol Exp Ther, 338(3):775-783.

[51]SuzukiK, TakeuchiO, SuzukiY, et al., 2019. Mechanisms of metformin’s anti-tumor activity against gemcitabine-resistant pancreatic adenocarcinoma. Int J Oncol, 54(2):764-772.

[52]TangGJ, GuoJF, ZhuYP, et al., 2018. Metformin inhibits ovarian cancer via decreasing H3K27 trimethylation. Int J Oncol, 52(6):1899-1911.

[53]TaniguchiK, KarinM, 2018. NF-κB, inflammation, immunity and cancer: coming of age. Nat Rev Immunol, 18(5):309-324.

[54]TaniguchiS, FujikiH, KobayashiH, et al., 1992. Effect of (-)-epigallocatechin gallate, the main constituent of green tea, on lung metastasis with mouse B16 melanoma cell lines. Cancer Lett, 65(1):51-54.

[55]TomicT, BottonT, CerezoM, et al., 2011. Metformin inhibits melanoma development through autophagy and apoptosis mechanisms. Cell Death Dis, 2(9):e199.

[56]ToyokuniS, OkamotoK, YodoiJ, et al., 1995. Persistent oxidative stress in cancer. FEBS Lett, 358(1):1-3.

[57]TrachoothamD, AlexandreJ, HuangP, 2009. Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat Rev Drug Discov, 8(7):579-591.

[58]WangL, LiK, LinXJ, et al., 2019. Metformin induces human esophageal carcinoma cell pyroptosis by targeting the miR-497/PELP1 axis. Cancer Lett, 450:22-31.

[59]WatanabeT, KuramochiH, TakahashiA, et al., 2012. Higher cell stiffness indicating lower metastatic potential in B16 melanoma cell variants and in (-)-epigallocatechin gallate-treated cells. J Cancer Res Clin Oncol, 138(5):859-866.

[60]WeiR, PensoNEC, HackmanRM, et al., 2019a. Epigallocatechin-3-gallate (EGCG) suppresses pancreatic cancer cell growth, invasion, and migration partly through the inhibition of Akt pathway and epithelial-mesenchymal transition: enhanced efficacy when combined with gemcitabine. Nutrients, 11(8):1856.

[61]WeiR, HackmanRM, WangYF, et al., 2019b. Targeting glycolysis with epigallocatechin-3-gallate enhances the efficacy of chemotherapeutics in pancreatic cancer cells and xenografts. Cancers, 11(10):1496.

[62]WoltersS, SchumacherB, 2013. Genome maintenance and transcription integrity in aging and disease. Front Genet, 4:19.

[63]WuY, LinY, LiuHJ, et al., 2008. Inhibition of invasion and up-regulation of E-cadherin expression in human malignant melanoma cell line A375 by (-)-epigallocatechin-3-gallate. J Huazhong Univ Sci Technolog Med Sci, 28(3):356-359.

[64]XuN, ZhuPP, LiangJ, et al., 2019. Label-free Raman spectroscopy monitoring of cytotoxic response induced by a telomerase inhibitor. Sens Actuat B Chem, 293:1-10.

[65]XuP, YanF, ZhaoYL, et al., 2020. Green tea polyphenol EGCG attenuates MDSCs-mediated immunosuppression through canonical and non-canonical pathways in a 4T1 murine breast cancer model. Nutrients, 12(4):1042.

[66]YamadaS, TsukamotoS, HuangYH, et al., 2016. Epigallocatechin-3-O-gallate up-regulates microRNA-let-7b expression by activating 67-kDa laminin receptor signaling in melanoma cells. Sci Rep, 6:19225.

[67]YangCS, WangX, LuG, et al., 2009. Cancer prevention by tea: animal studies, molecular mechanisms and human relevance. Nat Rev Cancer, 9(6):429-439.

[68]YuCX, JiaoY, XueJ, et al., 2017. Metformin sensitizes non-small cell lung cancer cells to an epigallocatechin-3-gallate (EGCG) treatment by suppressing the Nrf2/HO-1 signaling pathway. Int J Biol Sci, 13(12):1560-1569.

[69]YuH, KortylewskiM, PardollD, 2007. Crosstalk between cancer and immune cells: role of STAT3 in the tumour microenvironment. Nat Rev Immunol, 7(1):41-51.

[70]YuH, PardollD, JoveR, 2009. STATs in cancer inflammation and immunity: a leading role for STAT3. Nat Rev Cancer, 9(11):798-809.

[71]YuX, ZhouW, WangHM, et al., 2019. Transdermal metformin hydrochloride-loaded cubic phases: in silico formulation optimization, preparation, properties, and application for local treatment of melanoma. Drug Deliv, 26(1):376-383.

[72]ZhangJL, LeiZ, HuangZN, et al., 2016. Epigallocatechin-3-gallate (EGCG) suppresses melanoma cell growth and metastasis by targeting TRAF6 activity. Oncotarget, 7(48):79557-79571.

[73]ZhangYP, PengGY, HsuehEC, 2015. Abstract 1196: enhancement of anti-melanoma effect of BRAF and MEK inhibition bymetformin. Cancer Res, 75(15):1196.

[74]ZhouY, HilemanEO, PlunkettW, et al., 2003. Free radical stress in chronic lymphocytic leukemia cells and its role in cellular sensitivity to ROS-generating anticancer agents. Blood, 101(10):4098-4104.

[75]ZordokyBNM, BarkD, SoltysCL, et al., 2014. The anti-proliferative effect of metformin in triple-negative MDA-MB-231 breast cancer cells is highly dependent on glucose concentration: implications for cancer therapy and prevention. Biochim Biophys Acta, 1840(6):1943-1957.

[76]ZouG, BaiJ, LiDD, et al., 2019. Effect of metformin on the proliferation, apoptosis, invasion and autophagy of ovarian cancer cells. Exp Ther Med, 18(3):2086-2094.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE