Growth inhibition of luteolin on HepG2 cells is induced via p53 and Fas/Fas-ligand besides the TGF-β pathway

Yee,Su  Bog; Choi,Hye  Joung; Chung,Sang  Woon; Park,Dong  Heun; Sung,Bokyung; Chung,Hae  Young; Kim,Nam  Deuk

doi:10.3892/ijo.2015.3053

Growth inhibition of luteolin on HepG2 cells is induced via p53 and Fas/Fas-ligand besides the TGF-β pathway

Authors:
- Su Bog Yee
- Hye Joung Choi
- Sang Woon Chung
- Dong Heun Park
- Bokyung Sung
- Hae Young Chung
- Nam Deuk Kim
View Affiliations

Affiliations: Department of Clinical Laboratory Science, College of Nursing and Healthcare Sciences, Dong-Eui University, Busan 614-714, Republic of Korea, Department of Pharmacy, Molecular Inflammation Research Center for Aging Intervention, Pusan National University, Busan 609-735, Republic of Korea, School of Medicine, Korea University, Seoul 136-701, Republic of Korea
Published online on: June 18, 2015 https://doi.org/10.3892/ijo.2015.3053
Pages: 747-754

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

Flavonoids, a class of natural polyphenolic compounds, inhibit cell cycle progression and induce apoptosis. This study was performed to investigate the antiproliferative effect of luteolin, the flavonoid isolated from Ixeris sonchifolia Hance, and to elucidate the detailed apoptotic mechanism in HCC cells. According to the result of MTT assay luteolin possessed antiproliferative effect, and HepG2 cells were the most sensitive to luteolin. Propidium iodide staining, fluorescence activated cell sorting analysis, western blot analysis and RT-PCR were applied to compare the difference of apoptotic event between the two HCC cell lines, with wild-type p53 (HepG2) or not (Hep3B) based on time and concentration. The treatment of luteolin upregulated the expression levels of transforming growth factor β1 (TGF‑β1), p21WAF1/CIP1, p27KIP1, Smad4, and Fas in HCC cells. Thus, the expression of p21WAF1/CIP1 was controlled by another factor, such as TGF‑β1 in addition to p53, and notably the key factor might be p21WAF1/CIP1 in the remarkable switch to G1 cell cycle arrest in HepG2 cells rather than p27KIP1. Luteolin induced apoptotic cell death in Hep3B cells while caused G1 arrest in HepG2 cells. Taken together, we conclude that luteolin induces apoptosis from G1 arrest via three signaling pathways of TGF‑β1, p53, and Fas/Fas-ligand in HCC cells.

View Figures

View References

1	Chen D and Dou QP: Tea polyphenols and their roles in cancer prevention and chemotherapy. Int J Mol Sci. 9:1196–1206. 2008. View Article : Google Scholar
2	Berdowska I, Zieliński B, Fecka I, Kulbacka J, Saczko J and Gamian A: Cytotoxic impact of phenolics from Lamiaceae species on human breast cancer cells. Food Chem. 141:1313–1321. 2013. View Article : Google Scholar : PubMed/NCBI
3	Woodman OL and Chan EC: Vascular and anti-oxidant actions of flavonols and flavones. Clin Exp Pharmacol Physiol. 31:786–790. 2004. View Article : Google Scholar : PubMed/NCBI
4	Russo P, Del Bufalo A and Cesario A: Flavonoids acting on DNA topoisomerases: Recent advances and future perspectives in cancer therapy. Curr Med Chem. 19:5287–5293. 2012. View Article : Google Scholar : PubMed/NCBI
5	Guerra-Araiza C, Álvarez-Mejía AL, Sánchez-Torres S, Farfan-García E, Mondragón-Lozano R, Pinto-Almazán R and Salgado-Ceballos H: Effect of natural exogenous antioxidants on aging and on neurodegenerative diseases. Free Radic Res. 47:451–462. 2013. View Article : Google Scholar : PubMed/NCBI
6	Kritas SK, Saggini A, Varvara G, Murmura G, Caraffa A, Antinolfi P, Toniato E, Pantalone A, Neri G, Frydas S, et al: Luteolin inhibits mast cell-mediated allergic inflammation. J Biol Regul Homeost Agents. 27:955–959. 2013.
7	Sak K: Cytotoxicity of dietary flavonoids on different human cancer types. Pharmacogn Rev. 8:122–146. 2014. View Article : Google Scholar : PubMed/NCBI
8	Kaul TN, Middleton E Jr and Ogra PL: Antiviral effect of flavonoids on human viruses. J Med Virol. 15:71–79. 1985. View Article : Google Scholar : PubMed/NCBI
9	Kayser O, Kiderlen AF and Croft SL: Natural products as anti-parasitic drugs. Parasitol Res. 90(Suppl 2): S55–S62. 2003. View Article : Google Scholar
10	Lin YS, Tsai PH, Kandaswami CC, Cheng CH, Ke FC, Lee PP, Hwang JJ and Lee MT: Effects of dietary flavonoids, luteolin, and quercetin on the reversal of epithelial-mesenchymal transition in A431 epidermal cancer cells. Cancer Sci. 102:1829–1839. 2011. View Article : Google Scholar : PubMed/NCBI
11	López-Lázaro M: Flavonoids as anticancer agents: Structure-activity relationship study. Curr Med Chem Anticancer Agents. 2:691–714. 2002. View Article : Google Scholar
12	Chahar MK, Sharma N, Dobhal MP and Joshi YC: Flavonoids: A versatile source of anticancer drugs. Pharmacogn Rev. 5:1–12. 2011. View Article : Google Scholar : PubMed/NCBI
13	Kawaii S, Tomono Y, Katase E, Ogawa K and Yano M: Antiproliferative activity of flavonoids on several cancer cell lines. Biosci Biotechnol Biochem. 63:896–899. 1999. View Article : Google Scholar : PubMed/NCBI
14	Singh RP and Agarwal R: Natural flavonoids targeting deregulated cell cycle progression in cancer cells. Curr Drug Targets. 7:345–354. 2006. View Article : Google Scholar : PubMed/NCBI
15	Ramos S: Effects of dietary flavonoids on apoptotic pathways related to cancer chemoprevention. J Nutr Biochem. 18:427–442. 2007. View Article : Google Scholar : PubMed/NCBI
16	Pandurangan AK, Kumar SA, Dharmalingam P and Ganapasam S: Luteolin, a bioflavonoid inhibits azoxymethane-induced colon carcinogenesis: Involvement of iNOS and COX-2. Pharmacogn Mag. 10(Suppl 2): S306–S310. 2014. View Article : Google Scholar : PubMed/NCBI
17	Tofighi Z, Molazem M, Doostdar B, Taban P, Shahverdi AR, Samadi N and Yassa N: Antimicrobial activities of three medicinal plants and investigation of flavonoids of tripleurospermum disciforme. Iran J Pharm Res. 14:225–231. 2015.PubMed/NCBI
18	Li YC, Hung CF, Yeh FT, Lin JP and Chung JG: Luteolin-inhibited arylamineN-acetyltransferaseactivityandDNA-2-aminofluorene adduct in human and mouse leukemia cells. Food Chem Toxicol. 39:641–647. 2001. View Article : Google Scholar : PubMed/NCBI
19	Pérez-García F, Adzet T and Cañigueral S: Activity of artichoke leaf extract on reactive oxygen species in human leukocytes. Free Radic Res. 33:661–665. 2000. View Article : Google Scholar
20	Gebhardt R: Inhibition of cholesterol biosynthesis in HepG2 cells by artichoke extracts is reinforced by glucosidase pretreatment. Phytother Res. 16:368–372. 2002. View Article : Google Scholar : PubMed/NCBI
21	Wang TT, Wang SK, Huang GL and Sun GJ: Luteolin induced-growth inhibition and apoptosis of human esophageal squamous carcinoma cell line Eca109 cells in vitro. Asian Pac J Cancer Prev. 13:5455–5461. 2012. View Article : Google Scholar
22	Cai X, Lu W, Ye T, Lu M, Wang J, Huo J, Qian S, Wang X and Cao P: The molecular mechanism of luteolin-induced apoptosis is potentially related to inhibition of angiogenesis in human pancreatic carcinoma cells. Oncol Rep. 28:1353–1361. 2012.PubMed/NCBI
23	Lu J, Li G, He K, Jiang W, Xu C, Li Z, Wang H, Wang W, Wang H, Teng X, et al: Luteolin exerts a marked antitumor effect in cMet-overexpressing patient-derived tumor xenograft models of gastric cancer. J Transl Med. 13:422015. View Article : Google Scholar : PubMed/NCBI
24	Chiu FL and Lin JK: Downregulation of androgen receptor expression by luteolin causes inhibition of cell proliferation and induction of apoptosis in human prostate cancer cells and xenografts. Prostate. 68:61–71. 2008. View Article : Google Scholar
25	Chowdhury AR, Sharma S, Mandal S, Goswami A, Mukhopadhyay S and Majumder HK: Luteolin, an emerging anti-cancer flavonoid, poisons eukaryotic DNA topoisomerase I. Biochem J. 366:653–661. 2002. View Article : Google Scholar : PubMed/NCBI
26	Buening MK, Chang RL, Huang MT, Fortner JG, Wood AW and Conney AH: Activation and inhibition of benzo(a)pyrene and aflatoxin B1 metabolism in human liver microsomes by naturally occurring flavonoids. Cancer Res. 41:67–72. 1981.PubMed/NCBI
27	Ferriola PC, Cody V and Middleton E Jr: Protein kinase C inhibition by plant flavonoids. Kinetic mechanisms and structure-activity relationships. Biochem Pharmacol. 38:1617–1624. 1989. View Article : Google Scholar : PubMed/NCBI
28	Ko WG, Kang TH, Lee SJ, Kim YC and Lee BH: Effects of luteolin on the inhibition of proliferation and induction of apoptosis in human myeloid leukaemia cells. Phytother Res. 16:295–298. 2002. View Article : Google Scholar : PubMed/NCBI
29	Selvendiran K, Koga H, Ueno T, Yoshida T, Maeyama M, Torimura T, Yano H, Kojiro M and Sata M: Luteolin promotes degradation in signal transducer and activator of transcription 3 in human hepatoma cells: An implication for the antitumor potential of flavonoids. Cancer Res. 66:4826–4834. 2006. View Article : Google Scholar : PubMed/NCBI
30	Chang J, Hsu Y, Kuo P, Kuo Y, Chiang L and Lin C: Increase of Bax/ Bcl-XL ratio and arrest of cell cycle by luteolin in immortalized human hepatoma cell line. Life Sci. 76:1883–1893. 2005. View Article : Google Scholar : PubMed/NCBI
31	Villanueva A, García C, Paules AB, Vicente M, Megías M, Reyes G, de Villalonga P, Agell N, Lluís F, Bachs O, et al: Disruption of the antiproliferative TGF-beta signaling pathways in human pancreatic cancer cells. Oncogene. 17:1969–1978. 1998. View Article : Google Scholar : PubMed/NCBI
32	Yee SB, Lee JH, Chung HY, Im KS, Bae SJ, Choi JS and Kim ND: Inhibitory effects of luteolin isolated from Ixeris sonchifolia Hance on the proliferation of HepG2 human hepatocellular carcinoma cells. Arch Pharm Res. 26:151–156. 2003. View Article : Google Scholar : PubMed/NCBI
33	Derynck R, Jarrett JA, Chen EY, Eaton DH, Bell JR, Assoian RK, Roberts AB, Sporn MB and Goeddel DV: Human transforming growth factor-beta complementary DNA sequence and expression in normal and transformed cells. Nature. 316:701–705. 1985. View Article : Google Scholar : PubMed/NCBI
34	Hartwell LH and Kastan MB: Cell cycle control and cancer. Science. 266:1821–1828. 1994. View Article : Google Scholar : PubMed/NCBI
35	Polyak K, Kato JY, Solomon MJ, Sherr CJ, Massague J, Roberts JM and Koff A: p27^Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-beta and contact inhibition to cell cycle arrest. Genes Dev. 8:9–22. 1994. View Article : Google Scholar : PubMed/NCBI
36	Toyoshima H and Hunter T: p27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21. Cell. 78:67–74. 1994. View Article : Google Scholar : PubMed/NCBI
37	Massagué J: TGF-beta signal transduction. Annu Rev Biochem. 67:753–791. 1998. View Article : Google Scholar : PubMed/NCBI
38	Kim SG, Jong HS, Kim TY, Lee JW, Kim NK, Hong SH and Bang YJ: Transforming growth factor-beta 1 induces apoptosis through Fas ligand-independent activation of the Fas death pathway in human gastric SNU-620 carcinoma cells. Mol Biol Cell. 15:420–434. 2004. View Article : Google Scholar :
39	Depoortere F, Pirson I, Bartek J, Dumont JE and Roger PP: Transforming growth factor beta(1) selectively inhibits the cyclic AMP-dependent proliferation of primary thyroid epithelial cells by preventing the association of cyclin D3-cdk4 with nuclear p27(kip1). Mol Biol Cell. 11:1061–1076. 2000. View Article : Google Scholar : PubMed/NCBI
40	Park BJ, Park JI, Byun DS, Park JH and Chi SG: Mitogenic conversion of transforming growth factor-beta1 effect by oncogenic Ha-Ras-induced activation of the mitogen-activated protein kinase signaling pathway in human prostate cancer. Cancer Res. 60:3031–3038. 2000.PubMed/NCBI
41	Nakao A, Imamura T, Souchelnytskyi S, Kawabata M, Ishisaki A, Oeda E, Tamaki K, Hanai J, Heldin CH, Miyazono K, et al: TGF-beta receptor-mediated signalling through Smad2, Smad3 and Smad4. EMBO J. 16:5353–5362. 1997. View Article : Google Scholar : PubMed/NCBI
42	Zhang Q, Zhao XH and Wang ZJ: Cytotoxicity of flavones and flavonols to a human esophageal squamous cell carcinoma cell line (KYSE-510) by induction of G2/M arrest and apoptosis. Toxicol In Vitro. 23:797–807. 2009. View Article : Google Scholar : PubMed/NCBI
43	Kobayashi T, Nakata T and Kuzumaki T: Effect of flavonoids on cell cycle progression in prostate cancer cells. Cancer Lett. 176:17–23. 2002. View Article : Google Scholar : PubMed/NCBI
44	Walker A, Taylor ST, Hickman JA and Dive C: Germinal center-derived signals act with Bcl-2 to decrease apoptosis and increase clonogenicity of drug-treated human B lymphoma cells. Cancer Res. 57:1939–1945. 1997.PubMed/NCBI
45	Cory S, Vaux DL, Strasser A, Harris AW and Adams JM: Insights from Bcl-2 and Myc: Malignancy involves abrogation of apoptosis as well as sustained proliferation. Cancer Res. 59(Suppl): S1685–S1692. 1999.
46	Green DR and Evan GI: A matter of life and death. Cancer Cell. 1:19–30. 2002. View Article : Google Scholar : PubMed/NCBI
47	Crescenzi E and Palumbo G: Bcl-2 exerts a pRb-mediated cell cycle inhibitory function in HEC1B endometrial carcinoma cells. Gynecol Oncol. 81:184–192. 2001. View Article : Google Scholar : PubMed/NCBI
48	Attoub S, Hassan AH, Vanhoecke B, Iratni R, Takahashi T, Gaben AM, Bracke M, Awad S, John A, Kamalboor HA, et al: Inhibition of cell survival, invasion, tumor growth and histone deacetylase activity by the dietary flavonoid luteolin in human epithelioid cancer cells. Eur J Pharmacol. 651:18–25. 2011. View Article : Google Scholar