Antiproliferative and apoptotic effect of epigallocatechin-3-gallate on Ishikawa cells is accompanied by sex steroid receptor downregulation

Park,Seung  Bin; Bae,Jong  Woon; Kim,Jong  Min; Lee,Seung  Gee; Han,Myoungseok

doi:10.3892/ijmm.2012.1104

Antiproliferative and apoptotic effect of epigallocatechin-3-gallate on Ishikawa cells is accompanied by sex steroid receptor downregulation

Authors:
- Seung Bin Park
- Jong Woon Bae
- Jong Min Kim
- Seung Gee Lee
- Myoungseok Han
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, Dong-A University, College of Medicine, Busan 602-715, Republic of Korea, Department of Anatomy and Cell Biology, Dong-A University, College of Medicine, Busan 602-715, Republic of Korea
Published online on: August 20, 2012 https://doi.org/10.3892/ijmm.2012.1104
Pages: 1211-1218

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Endometrial cancer is a significant malignancy in developed countries. Unopposed estrogen stimulation is considered as an important risk factor for endometrial cancer. Epigallocathechin-3-gallate (EGCG), biological active component of green tea, inhibits cancer cell proliferation. However, it is unknown whether EGCG has anticancer effects on endometrial cancer and what the molecular mechanism(s) are. We investigated the anticancer effects of EGCG on a human endometrial adenocarcinoma cell line (Ishikawa cells) with or without 17β-estradiol (E2) treatment. Cell proliferation assay was performed using 3-(4,5-dimethylthiaxol-2-yi)-2,5-diphenyltetraxolium bromide (MTT). The cell cycle was determined by flow cytometry and real-time analysis of cyclin and cdk genes. The apoptosis was measured by Annexin V-PI staining and real-time analysis of bcl-2, Bax and caspase genes. The MAPK signal, Akt and caspase-3 were determined by immunoblotting. Decreased estrogen and progesterone receptor expression was observed in EGCG-treated Ishikawa cells, and decreased MAPK signals and phospho-Akt were observed as well. EGCG caused the arrest of cells in the G0/G1 phase of the cell cycle. This compound interfered with Akt activation and MAPK signals, and increased apoptosis signals leading to a controlled caspases, Bcl-2, Bax genes and protein expression. Taken together, EGCG inhibits cell proliferation and induces apoptosis through Akt and MAPK signals. These findings suggest that EGCG may exert growth-inhibitory and apoptosis-inducing effects on endometrial cancer cells, accompanied by decreased estrogen and progesterone receptor. EGCG may have future clinical implications with respect to the development of novel approaches as an adjuvant therapy in endometrial cancer.

View Figures

View References

1.	A DollM AbalM RigauNovel molecular profiles of endometrial cancer-new light through old windowsJ Steroid Biochem Mol Biol108221229200810.1016/j.jsbmb.2007.09.02018061438
2.	G AlbrektsenI HeuchS TretliG KvaleIs the risk of cancer of the corpus uteri reduced by a recent pregnancy? A prospective study of 765,756 Norwegian womenInt J Cancer61485490199510.1002/ijc.29106104107759154
3.	A KalandidiA TzonouL LipworthI GamatsiD FilippaD TrichopoulosA case-control study of endometrial cancer in relation to reproductive, somatometric, and life-style variablesOncology53354359199610.1159/0002275878784467
4.	D GradyT GebretsadikK KerlikowskeV ErnsterD PetittiHormone replacement therapy and endometrial cancer risk: a meta-analysisObstet Gynecol85304313199510.1016/0029-7844(94)00383-O7824251
5.	A AkhmedkhanovA Zeleniuch-JacquotteP TonioloRole of exogenous and endogenous hormones in endometrial cancer: review of the evidence and research perspectivesAnn NY Acad Sci943296315200110.1111/j.1749-6632.2001.tb03811.x11594550
6.	CF HolinkaY AnzaiH HataN KimmelH KuramotoE GurpideProliferation and responsiveness to estrogen of human endometrial cancer cells under serum-free culture conditionsCancer Res493297330119892720684
7.	A BouskineM NeboutB MograbiF Brucker-DavisC RogerP FenichelEstrogens promote human testicular germ cell cancer through a membrane-mediated activation of extracellular regulated kinase and protein kinase AEndocrinology149565573200810.1210/en.2007-1318
8.	G CastoriaA MigliaccioA BilancioPI3-kinase in concert with Src promotes the S-phase entry of oestradiol-stimulated MCF-7 cellsEMBO J2060506059200110.1093/emboj/20.21.605011689445
9.	JA RobertsonY FarnellLS LindahlNH IngEstradiol up-regulates estrogen receptor messenger ribonucleic acid in endometrial carcinoma (Ishikawa) cells by stabilizing the messageJ Mol Endocrinol29125135200210.1677/jme.0.029012512200234
10.	BJ CheskisJ GregerN CoochMNAR plays an important role in ERa activation of Src/MAPK and PI3K/Akt signaling pathwaysSteroids73901905200810.1016/j.steroids.2007.12.02818261753
11.	BM WolpinRJ MayerSystemic treatment of colorectal cancerGastroenterology13412961310200810.1053/j.gastro.2008.02.09818471507
12.	H FujikiM SuganumaS OkabeJapanese green tea as a cancer preventive in humansNutr Rev54S67S70199610.1111/j.1753-4887.1996.tb03821.x9110578
13.	MH PanYS ChiouYJ WangCT HoJK LinMultistage carcinogenesis process as molecular targets in cancer chemoprevention by epicatechin-3-gallateFood Funct2101110201110.1039/c0fo00174k21779554
14.	T WebbGreen tea experiments in lab, clinic yield mixed resultsJ Natl Cancer Inst9210381039200010.1093/jnci/92.13.103810880545
15.	Y CaoR CaoAngiogenesis inhibited by drinking teaNature398381199910.1038/1879310201368
16.	M YokoyamaM NoguchiY NakaoA PaterT IwasakaThe tea polyphenol, (−)-epigallocatechin gallate effects on growth, apoptosis, and telomerase activity in cervical cell linesGynecol Oncol921972042004
17.	G ZhangV GurtuSR KainG YanEarly detection of apoptosis using a fluorescent conjugate of annexin VBiotechniques2352553119979298227
18.	DM ParkinF BrayJ FerlayP PisaniGlobal cancer statistics, 2002CA Cancer J Clin5574108200510.3322/canjclin.55.2.74
19.	V BeralD BullG ReevesEndometrial cancer and hormone-replacement therapy in the Million Women StudyLancet36515431551200510.1016/S0140-6736(05)66455-015866308
20.	LP LovelyKB Appa RaoY GuiBA LesseyCharacterization of androgen receptors in a well-differentiated endometrial adenocarcinoma cell line (Ishikawa)J Steroid Biochem Mol Biol74235241200010.1016/S0960-0760(00)00127-811162929
21.	MP SnijdersAF de GoeijMJ Debets-Te BaertsMJ RouschJ KoudstaalFT BosmanImmunocytochemical analysis of oestrogen receptors and progesterone receptors in the human uterus throughout the menstrual cycle and after the menopauseJ Reprod Fertil94363371199210.1530/jrf.0.0940363
22.	M ShimizuA DeguchiJT LimH MoriwakiL KopelovichIB Weinstein(−)-Epigallocatechin gallate and polyphenon E inhibit growth and activation of the epidermal growth factor receptor and human epidermal growth factor receptor-2 signaling pathways in human colon cancer cellsClin Cancer Res11273527462005
23.	N KhanF AfaqM SaleemN AhmadH MukhtarTargeting multiple signaling pathways by green tea polyphenol (−)-epigallocatechin-3-gallateCancer Res66250025052006
24.	N AhmadDK FeyesAL NieminenR AgarwalH MukhtarGreen tea constituent epigallocatechin-3-gallate and induction of apoptosis and cell cycle arrest in human carcinoma cellsJ Natl Cancer Inst8918811886199710.1093/jnci/89.24.18819414176
25.	CS YangP MaliakalX MengInhibition of carcinogenesis by teaAnnu Rev Pharmacol Toxicol422554200210.1146/annurev.pharmtox.42.082101.15430911807163
26.	N AhmadP ChengH MukhtarCell cycle dysregulation by green tea polyphenol epigallocatechin-3-gallateBiochem Biophys Res Commun275328334200010.1006/bbrc.2000.329710964666
27.	X LiuDY ZhangW ZhangX ZhaoC YuanF YeThe effect of green tea extract and EGCG on the signaling network in squamous cell carcinomaNutr Cancer63466475201110.1080/01635581.2011.53290121391127
28.	WS AhnJ YooSW HuhProtective effects of green tea extracts (polyphenon E and EGCG) on human cervical lesionsEur J Cancer Prev12383390200310.1097/00008469-200310000-0000714512803
29.	A JatoiN EllisonPA BurchA phase II trial of green tea in the treatment of patients with androgen independent metastatic prostate carcinomaCancer9714421446200310.1002/cncr.1120012627508