Isoliquiritigenin inhibits the proliferation of human renal carcinoma Caki cells through the ROS-mediated 
regulation of the Jak2/STAT3 pathway

Kim,Do-Hee; Park,Ji  Eun; Chae,In  Gyeong; Park,Geumi; Lee,Sooyeun; Chun,Kyung-Soo

doi:10.3892/or.2017.5677

Isoliquiritigenin inhibits the proliferation of human renal carcinoma Caki cells through the ROS-mediated regulation of the Jak2/STAT3 pathway

Authors:
- Do-Hee Kim
- Ji Eun Park
- In Gyeong Chae
- Geumi Park
- Sooyeun Lee
- Kyung-Soo Chun
View Affiliations

Affiliations: College of Pharmacy, Seoul National University, Seoul 151-742, Republic of Korea, College of Pharmacy, Keimyung University, Daegu 704-701, Republic of Korea
Published online on: May 30, 2017 https://doi.org/10.3892/or.2017.5677
Pages: 575-583

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

Isoliquiritigenin (ISL) is a flavonoid with chalcone structure that has been noted in licorice and shallot, which are generally used in traditional Chinese medicine. ISL has demonstrated various pharmacological effects including antioxidant, anti-inflammatory and antitumor activity. However, the molecular mechanisms underlying the anticancer effects of ISL remain poorly understood. The present study revealed that ISL significantly decreased viability and induced apoptosis in human renal carcinoma Caki cells. The ISL-induced apoptosis was associated with the cleavage of caspase-9, -7 and -3, and that of PARP. Moreover, ISL increased the expression of pro-apoptotic protein Bax and diminished the expression of anti-apoptotic protein Bcl-2, and Bcl-xl, thereby increasing cytochrome c release. Treatment of cells with ISL also induced the expression of p53 through downregulation of murine double minute 2 (Mdm2). Furthermore, ISL generated reactive oxygen species (ROS), and pretreatment with ROS scavenger N-acetyl cysteine (NAC) and NADPH oxidase inhibitor diphenyleneiodonium abrogated the ISL-induced apoptosis. One of the key oncogenic signaling pathways is mediated through signal transducer and activator of transcription 3 (STAT3), which promotes abnormal cell proliferation. Incubation of cells with ISL markedly diminished phosphorylation and DNA binding activity of STAT3, and reduced expression of STAT3 responsive gene products, such as cyclin D1 and D2. ISL also attenuated constitutive phosphorylation of upstream kinase, Janus-activated kinase 2 (Jak2). Pretreatment with NAC abrogated the inhibitory effect of ISL on activation of STAT3 and blocked the cleavage of caspase-9, -7 and -3, and that of PARP in Caki cells. Taken together, the present study provides the first report that ISL induces apoptosis in Caki cells via generation of ROS, which causes induction of p53 and inhibition of the STAT3 signaling pathway.

View Figures

View References

1	Grimm MO, Wolff I, Zastrow S, Fröhner M and Wirth M: Advances in renal cell carcinoma treatment. Ther Adv Urol. 2:11–17. 2010. View Article : Google Scholar : PubMed/NCBI
2	De P, Otterstatter MC, Semenciw R, Ellison LF, Marrett LD and Dryer D: Trends in incidence, mortality, and survival for kidney cancer in Canada, 1986–2007. Cancer Causes Control. 25:1271–1281. 2014. View Article : Google Scholar : PubMed/NCBI
3	Motzer RJ, Michaelson MD, Redman BG, Hudes GR, Wilding G, Figlin RA, Ginsberg MS, Kim ST, Baum CM, DePrimo SE, et al: Activity of SU11248, a multitargeted inhibitor of vascular endothelial growth factor receptor and platelet-derived growth factor receptor, in patients with metastatic renal cell carcinoma. J Clin Oncol. 24:16–24. 2006. View Article : Google Scholar : PubMed/NCBI
4	Darnell JE Jr: STATs and gene regulation. Science. 277:1630–1635. 1997. View Article : Google Scholar : PubMed/NCBI
5	Dhir R, Ni Z, Lou W, DeMiguel F, Grandis JR and Gao AC: Stat3 activation in prostatic carcinomas. Prostate. 51:241–246. 2002. View Article : Google Scholar : PubMed/NCBI
6	Huang M, Page C, Reynolds RK and Lin J: Constitutive activation of stat 3 oncogene product in human ovarian carcinoma cells. Gynecol Oncol. 79:67–73. 2000. View Article : Google Scholar : PubMed/NCBI
7	Proietti CJ, Rosemblit C, Beguelin W, Rivas MA, Díaz Flaqué MC, Charreau EH, Schillaci R and Elizalde PV: Activation of Stat3 by heregulin/ErbB-2 through the co-option of progesterone receptor signaling drives breast cancer growth. Mol Cell Biol. 29:1249–1265. 2009. View Article : Google Scholar : PubMed/NCBI
8	Guo C, Yang G, Khun K, Kong X, Levy D, Lee P and Melamed J: Activation of Stat3 in renal tumors. Am J Transl Res. 1:283–290. 2009.PubMed/NCBI
9	Yu H, Lee H, Herrmann A, Buettner R and Jove R: Revisiting STAT3 signalling in cancer: New and unexpected biological functions. Nat Rev Cancer. 14:736–746. 2014. View Article : Google Scholar : PubMed/NCBI
10	Bowman T, Garcia R, Turkson J and Jove R: STATs in oncogenesis. Oncogene. 19:2474–2488. 2000. View Article : Google Scholar : PubMed/NCBI
11	Johnston PA and Grandis JR: STAT3 signaling: Anticancer strategies and challenges. Mol Interv. 11:18–26. 2011. View Article : Google Scholar : PubMed/NCBI
12	Carpenter RL and Lo HW: STAT3 target genes relevant to human cancers. Cancers. 6:897–925. 2014. View Article : Google Scholar : PubMed/NCBI
13	Portt L, Norman G, Clapp C, Greenwood M and Greenwood MT: Anti-apoptosis and cell survival: A review. Biochim Biophys Acta. 1813:238–259. 2011. View Article : Google Scholar : PubMed/NCBI
14	Riedl SJ and Salvesen GS: The apoptosome: Signalling platform of cell death. Nat Rev Mol Cell Biol. 8:405–413. 2007. View Article : Google Scholar : PubMed/NCBI
15	Sinha K, Das J, Pal PB and Sil PC: Oxidative stress: The mitochondria-dependent and mitochondria-independent pathways of apoptosis. Arch Toxicol. 87:1157–1180. 2013. View Article : Google Scholar : PubMed/NCBI
16	Cerna D, Li H, Flaherty S, Takebe N, Coleman CN and Yoo SS: Inhibition of nicotinamide phosphoribosyltransferase (NAMPT) activity by small molecule GMX1778 regulates reactive oxygen species (ROS)-mediated cytotoxicity in a p53- and nicotinic acid phosphoribosyltransferase1 (NAPRT1)-dependent manner. J Biol Chem. 287:22408–22417. 2012. View Article : Google Scholar : PubMed/NCBI
17	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
18	Kampa M, Nifli AP, Notas G and Castanas E: Polyphenols and cancer cell growth. Rev Physiol Biochem Pharmacol. 159:79–113. 2007.PubMed/NCBI
19	Aida K, Tawata M, Shindo H, Onaya T, Sasaki H, Yamaguchi T, Chin M and Mitsuhashi H: Isoliquiritigenin: A new aldose reductase inhibitor from Glycyrrhizae Radix. Planta Med. 56:254–258. 1990. View Article : Google Scholar : PubMed/NCBI
20	Choi YH, Bae JK, Chae HS, Choi YO, Nhoek P, Choi JS and Chin YW: Isoliquiritigenin ameliorates dextran sulfate sodium-induced colitis through the inhibition of MAPK pathway. Int Immunopharmacol. 31:223–232. 2016. View Article : Google Scholar : PubMed/NCBI
21	Gaur R, Yadav KS, Verma RK, Yadav NP and Bhakuni RS: In vivo anti-diabetic activity of derivatives of isoliquiritigenin and liquiritigenin. Phytomedicine. 21:415–422. 2014. View Article : Google Scholar : PubMed/NCBI
22	Jung SK, Lee MH, Lim DY, Kim JE, Singh P, Lee SY, Jeong CH, Lim TG, Chen H, Chi YI, et al: Isoliquiritigenin induces apoptosis and inhibits xenograft tumor growth of human lung cancer cells by targeting both wild type and L858R/T790M mutant EGFR. J Biol Chem. 289:35839–35848. 2014. View Article : Google Scholar : PubMed/NCBI
23	Auyeung KK and Ko JK: Novel herbal flavonoids promote apoptosis but differentially induce cell cycle arrest in human colon cancer cell. Invest New Drugs. 28:1–13. 2010. View Article : Google Scholar : PubMed/NCBI
24	Jung JI, Chung E, Seon MR, Shin HK, Kim EJ, Lim SS, Chung WY, Park KK and Park JH: Isoliquiritigenin (ISL) inhibits ErbB3 signaling in prostate cancer cells. Biofactors. 28:159–168. 2006. View Article : Google Scholar : PubMed/NCBI
25	Lai PH, Li KT, Hsu SS, Hsiao CC, Yip CW, Ding S, Yeh LR and Pan HB: Pyogenic brain abscess: Findings from in vivo 1.5-T and 11.7-T in vitro proton MR spectroscopy. AJNR Am J Neuroradiol. 26:279–288. 2005.PubMed/NCBI
26	Li Y, Zhao H, Wang Y, Zheng H, Yu W, Chai H, Zhang J, Falck JR, Guo AM, Yue J, et al: Isoliquiritigenin induces growth inhibition and apoptosis through downregulating arachidonic acid metabolic network and the deactivation of PI3K/Akt in human breast cancer. Toxicol Appl Pharmacol. 272:37–48. 2013. View Article : Google Scholar : PubMed/NCBI
27	Park I, Park KK, Park JH and Chung WY: Isoliquiritigenin induces G2 and M phase arrest by inducing DNA damage and by inhibiting the metaphase/anaphase transition. Cancer Lett. 277:174–181. 2009. View Article : Google Scholar : PubMed/NCBI
28	Wang Z, Wang N, Han S, Wang D, Mo S, Yu L, Huang H, Tsui K, Shen J and Chen J: Dietary compound isoliquiritigenin inhibits breast cancer neoangiogenesis via VEGF/VEGFR-2 signaling pathway. PLoS One. 8:e685662013. View Article : Google Scholar : PubMed/NCBI
29	Yamazaki S, Morita T, Endo H, Hamamoto T, Baba M, Joichi Y, Kaneko S, Okada Y, Okuyama T, Nishino H, et al: Isoliquiritigenin suppresses pulmonary metastasis of mouse renal cell carcinoma. Cancer Lett. 183:23–30. 2002. View Article : Google Scholar : PubMed/NCBI
30	Chae IG, Kim DH, Kundu J, Jeong CH, Kundu JK and Chun KS: Generation of ROS by CAY10598 leads to inactivation of STAT3 signaling and induction of apoptosis in human colon cancer HCT116 cells. Free Radic Res. 48:1311–1321. 2014. View Article : Google Scholar : PubMed/NCBI
31	Kim DH, Park KW, Chae IG, Kundu J, Kim EH, Kundu JK and Chun KS: Carnosic acid inhibits STAT3 signaling and induces apoptosis through generation of ROS in human colon cancer HCT116 cells. Mol Carcinog. 55:1096–1110. 2016. View Article : Google Scholar : PubMed/NCBI
32	Indran IR, Tufo G, Pervaiz S and Brenner C: Recent advances in apoptosis, mitochondria and drug resistance in cancer cells. Biochim Biophys Acta. 1807:735–745. 2011. View Article : Google Scholar : PubMed/NCBI
33	Simon HU, Haj-Yehia A and Levi-Schaffer F: Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis. 5:415–418. 2000. View Article : Google Scholar : PubMed/NCBI
34	Bromberg J: Stat proteins and oncogenesis. J Clin Invest. 109:1139–1142. 2002. View Article : Google Scholar : PubMed/NCBI
35	Aznar S, Valerón PF, del Rincon SV, Pérez LF, Perona R and Lacal JC: Simultaneous tyrosine and serine phosphorylation of STAT3 transcription factor is involved in Rho A GTPase oncogenic transformation. Mol Biol Cell. 12:3282–3294. 2001. View Article : Google Scholar : PubMed/NCBI
36	Bishayee A and Sethi G: Bioactive natural products in cancer prevention and therapy: Progress and promise. Semin Cancer Biol 40–41. 1–3. 2016. View Article : Google Scholar
37	Arumuggam N, Bhowmick NA and Rupasinghe HP: A Review: Phytochemicals Targeting JAK/STAT Signaling and IDO Expression in Cancer. Phytother Res. 29:805–817. 2015. View Article : Google Scholar : PubMed/NCBI
38	Lue HW, Cole B, Rao SA, Podolak J, Van Gaest A, King C, Eide CA, Wilmot B, Xue C, Spellman PT, et al: Src and STAT3 inhibitors synergize to promote tumor inhibition in renal cell carcinoma. Oncotarget. 6:44675–44687. 2015.PubMed/NCBI
39	Um HJ, Min KJ, Kim DE and Kwon TK: Withaferin A inhibits JAK/STAT3 signaling and induces apoptosis of human renal carcinoma Caki cells. Biochem Biophys Res Commun. 427:24–29. 2012. View Article : Google Scholar : PubMed/NCBI
40	Chin YW, Jung HA, Liu Y, Su BN, Castoro JA, Keller WJ, Pereira MA and Kinghorn AD: Anti-oxidant constituents of the roots and stolons of licorice (Glycyrrhiza glabra). J Agric Food Chem. 55:4691–4697. 2007. View Article : Google Scholar : PubMed/NCBI
41	Kakegawa H, Matsumoto H and Satoh T: Inhibitory effects of some natural products on the activation of hyaluronidase and their anti-allergic actions. Chem Pharm Bull. 40:1439–1442. 1992. View Article : Google Scholar : PubMed/NCBI
42	Kang SW, Choi JS, Choi YJ, Bae JY, Li J, Kim DS, Kim JL, Shin SY, Lee YJ and Kwun IS: Licorice isoliquiritigenin dampens angiogenic activity via inhibition of MAPK-responsive signaling pathways leading to induction of matrix metalloproteinases. J Nutr Biochem. 21:55–65. 2010. View Article : Google Scholar : PubMed/NCBI
43	Kape R, Parniske M, Brandt S and Werner D: Isoliquiritigenin, a strong nod gene- and glyceollin resistance-inducing flavonoid from soybean root exudate. Appl Environ Microbiol. 58:1705–1710. 1992.PubMed/NCBI
44	Deponte M: Glutathione catalysis and the reaction mechanisms of glutathione-dependent enzymes. Biochim Biophys Acta. 1830:3217–3266. 2013. View Article : Google Scholar : PubMed/NCBI
45	Bromberg J and Darnell JE Jr: The role of STATs in transcriptional control and their impact on cellular function. Oncogene. 19:2468–2473. 2000. View Article : Google Scholar : PubMed/NCBI
46	Heidelberger S, Zinzalla G, Antonow D, Essex S, Basu BP, Palmer J, Husby J, Jackson PJ, Rahman KM, Wilderspin AF, et al: Investigation of the protein alkylation sites of the STAT3:STAT3 inhibitor Stattic by mass spectrometry. Bioorg Med Chem Lett. 23:4719–4722. 2013. View Article : Google Scholar : PubMed/NCBI
47	Li L, Cheung SH, Evans EL and Shaw PE: Modulation of gene expression and tumor cell growth by redox modification of STAT3. Cancer Res. 70:8222–8232. 2010. View Article : Google Scholar : PubMed/NCBI
48	Park KW, Kundu J, Chae IG, Kim DH, Yu MH, Kundu JK and Chun KS: Carnosol induces apoptosis through generation of ROS and inactivation of STAT3 signaling in human colon cancer HCT116 cells. Int J Oncol. 44:1309–1315. 2014.PubMed/NCBI
49	Sinha P and Ostrand-Rosenberg S: Myeloid-derived suppressor cell function is reduced by Withaferin A, a potent and abundant component of Withania somnifera root extract. Cancer Immunol Immunother. 62:1663–1673. 2013. View Article : Google Scholar : PubMed/NCBI
50	Sullivan LB and Chandel NS: Mitochondrial reactive oxygen species and cancer. Cancer Metab. 2:172014. View Article : Google Scholar : PubMed/NCBI
51	Pan MH, Sin YH, Lai CS, Wang YJ, Lin JK, Wang M and Ho CT: Induction of apoptosis by 1-(2-hydroxy-5-methylphenyl)-3-phenyl-1,3-propanedione through reactive oxygen species production, GADD153 expression, and caspases activation in human epidermoid carcinoma cells. J Agric Food Chem. 53:9039–9049. 2005. View Article : Google Scholar : PubMed/NCBI
52	Li D, Ueta E, Kimura T, Yamamoto T and Osaki T: Reactive oxygen species (ROS) control the expression of Bcl-2 family proteins by regulating their phosphorylation and ubiquitination. Cancer Sci. 95:644–650. 2004. View Article : Google Scholar : PubMed/NCBI
53	Schumacker PT: Reactive oxygen species in cancer cells: Live by the sword, die by the sword. Cancer Cell. 10:175–176. 2006. View Article : Google Scholar : PubMed/NCBI
54	Jiang YQ, Zhou ZX and Ji YL: Suppression of EGFR-STAT3 signaling inhibits tumorigenesis in a lung cancer cell line. Int J Clin Exp Med. 7:2096–2099. 2014.PubMed/NCBI