RL71, a second-generation curcumin analog, induces apoptosis and downregulates Akt in ER-negative breast cancer cells

Yadav,Babasaheb; Taurin,Sebastien; Larsen,Lesley; Rosengren,Rhonda   J.

doi:10.3892/ijo.2012.1521

RL71, a second-generation curcumin analog, induces apoptosis and downregulates Akt in ER-negative breast cancer cells

Authors:
- Babasaheb Yadav
- Sebastien Taurin
- Lesley Larsen
- Rhonda J. Rosengren
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Affiliations: Department of Pharmacology and Toxicology, University of Otago, Dunedin, New Zealand, Department of Chemistry, University of Otago, Dunedin, New Zealand
Published online on: June 15, 2012 https://doi.org/10.3892/ijo.2012.1521
Pages: 1119-1127

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Abstract

There is a need for the development of new, safe and efficacious drug therapies for the treatment of estrogen receptor (ER)-negative breast cancers. RL71 is a second-generation curcumin analog that exhibits potent cytotoxicity towards a variety of ER-negative breast cancer cells. Therefore, we have further examined the mechanism of this anticancer activity in three different ER-negative breast cancer cell lines. The mechanistic studies demonstrated that RL71 (1 µM) induced cell cycle arrest in the G2/M phase of the cell cycle. Moreover, RL71 (1 µM) caused 35% of SKBr3 cells to undergo apoptosis after 48 h and this effect was time-dependent. This correlated with an increase in cleaved caspase-3 as shown by western blotting. RL71 (1 µM) also decreased HER2/neu phosphorylation and increased p27 in SKBr3 cells. While in MDA-MB-231 and MDA-MB-468 cells RL71 (1 µM) significantly decreased Akt phosphorylation and transiently increased the stress kinases JNK1/2 and p38 MAPK. In addition, RL71 exhibited anti-angiogenic potential in vitro as it inhibited HUVEC cell migration and the ability of these cells to form tube-like networks. RL71 (8.5 mg/kg) was also orally bioavailable as it produced a peak plasma concentration of 0.405 µg/ml, 5 min after oral drug administration. Thus, our findings provide evidence that RL71 has potent anticancer activity and has potential to be further developed as a drug for the treatment of ER-negative breast cancer.

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1.	Parl FF, Schmidt BP, Dupont WD and Wagner RK: Prognostic significance of estrogen receptor status in breast cancer in relation to tumor stage, axillary node metastasis, and histopathologic grading. Cancer. 54:2237–2242. 1984. View Article : Google Scholar : PubMed/NCBI
2.	Doane AS, Danso M, Lal P, Donaton M, Zhang L, Hudis C and Gerald WL: An estrogen receptor-negative breast cancer subset characterized by a hormonally regulated transcriptional program and response to androgen. Oncogene. 25:3994–4008. 2006. View Article : Google Scholar
3.	Carey LA, Dees EC, Sawyer L, et al: The triple negative paradox: primary tumor chemosensitivity of breast cancer subtypes. Clin Cancer Res. 13:2329–2334. 2007. View Article : Google Scholar : PubMed/NCBI
4.	Sorlie T, Perou CM, Tibshirani R, et al: Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci USA. 98:10869–10874. 2001. View Article : Google Scholar : PubMed/NCBI
5.	Kaplan HG and Malmgren JA: Impact of triple negative phenotype on breast cancer prognosis. Breast J. 14:456–463. 2008. View Article : Google Scholar : PubMed/NCBI
6.	Anand P, Thomas Sherin G, Kunnumakkara Ajaikumar B, et al: Biological activities of curcumin and its analogues (Congeners) made by man and Mother Nature. Biochem Pharmacol. 76:1590–1611. 2008. View Article : Google Scholar : PubMed/NCBI
7.	Cheng A-L, Hsu C-H, Lin J-K, et al: Phase I clinical trial of curcumin, a chemopreventive agent, in patients with high-risk or pre-malignant lesions. Anticancer Res. 21:2895–2900. 2001.PubMed/NCBI
8.	Inano H, Onoda M, Inafuku N, et al: Chemoprevention by curcumin during the promotion stage of tumorigenesis of mammary gland in rats irradiated with gamma-rays. Carcinogenesis. 20:1011–1018. 1999. View Article : Google Scholar : PubMed/NCBI
9.	Pereira MA, Grubbs CJ, Barnes LH, et al: Effects of the phytochemicals, curcumin and quercetin, upon azoxymethane-induced colon cancer and 7,12-dimethylbenz[a]anthracene-induced mammary cancer in rats. Carcinogenesis. 17:1305–1311. 1996.PubMed/NCBI
10.	Schaaf C, Shan B, Buchfelder M, et al: Curcumin acts as anti-tumorigenic and hormone-suppressive agent in murine and human pituitary tumour cells in vitro and in vivo. Endocr Relat Cancer. 16:1339–1350. 2009. View Article : Google Scholar : PubMed/NCBI
11.	Singletary K, MacDonald C, Wallig M and Fisher C: Inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumorigenesis and DMBA-DNA adduct formation by curcumin. Cancer Lett. 103:137–141. 1996.
12.	Chiu T-L and Su C-C: Curcumin inhibits proliferation and migration by increasing the Bax to Bcl-2 ratio and decreasing NF-κBp65 expression in breast cancer MDA-MB-231 cells. Int J Mol Med. 23:469–475. 2009.PubMed/NCBI
13.	Kang HJ, Lee SH, Price JE and Kim LS: Curcumin suppresses the paclitaxel-induced nuclear factor-κB in breast cancer cells and potentiates the growth inhibitory effect of paclitaxel in a breast cancer nude mice model. Breast J. 15:223–229. 2009.
14.	Liu Q, Loo WTY, Sze SCW and Tong Y: Curcumin inhibits cell proliferation of MDA-MB-231 and BT-483 breast cancer cells mediated by down-regulation of NFκB, cyclinD and MMP-1 transcription. Phytomed. 16:916–922. 2009.PubMed/NCBI
15.	Prasad CP, Rath G, Mathur S, Bhatnagar D and Ralhan R: Potent growth suppressive activity of curcumin in human breast cancer cells: Modulation of Wnt/beta-catenin signaling. Chem Biol Interact. 181:263–271. 2009. View Article : Google Scholar : PubMed/NCBI
16.	Rowe DL, Ozbay T, O’Regan RM and Nahta R: Modulation of the BRCA1 protein and induction of apoptosis in triple negative breast cancer cell lines by the polyphenolic compound curcumin. Breast Cancer Basic Clin Res. 3:61–75. 2009.PubMed/NCBI
17.	Somers-Edgar TJ, Scandlyn MJ, Stuart EC, Le Nedelec MJ, Valentine SP and Rosengren RJ: The combination of epigallocatechin gallate and curcumin suppresses ERalpha-breast cancer cell growth in vitro and in vivo. Int J Cancer. 122:1966–1971. 2008. View Article : Google Scholar : PubMed/NCBI
18.	Wu X and Wu K: Antiproliferative effect of curcumin on human breast cancer of MCF-7 cells. Di-San Junyi Daxue Xuebao. 28:1870–1872. 2006.
19.	Liang G, Shao L, Wang Y, et al: Exploration and synthesis of curcumin analogues with improved structural stability both in vitro and in vivo as cytotoxic agents. Bioorg Med Chem. 17:2623–2631. 2009. View Article : Google Scholar : PubMed/NCBI
20.	Markaverich BM, Schauweker TH, Gregory RR, Varma M, Kittrell FS, Medina D and Varma RS: Nuclear type II sites and malignant cell proliferation: inhibition by 2,6-bis-benzylidenecyclohexanones. Cancer Res. 52:2482–2488. 1992.PubMed/NCBI
21.	Adams BK, Ferstl EM, Davis MC, et al: Synthesis and biological evaluation of novel curcumin analogs as anti-cancer and anti-angiogenesis agents. Bioorg Med Chem. 12:3871–3883. 2004. View Article : Google Scholar : PubMed/NCBI
22.	Adams Brian K, Cai J, Armstrong J, et al: EF24, a novel synthetic curcumin analog, induces apoptosis in cancer cells via a redox-dependent mechanism. Anticancer Drugs. 16:263–275. 2005.PubMed/NCBI
23.	Yadav B, Taurin S, Rosengren RJ, Schumacher M, Diederich M, Somers-Edgar TJ and Larsen L: Synthesis and cytotoxic potential of heterocyclic cyclohexanone analogues of curcumin. Bioorg Med Chem. 18:6701–6707. 2010. View Article : Google Scholar : PubMed/NCBI
24.	Skehan P, Storeng R, Scudiero D, et al: New colorimetric cytotoxicity assay for anti-cancer drug screening. J Natl Cancer Inst. 82:1107–1112. 1990. View Article : Google Scholar : PubMed/NCBI
25.	Somers-Edgar TJ, Taurin S, Larsen L, Chandramouli A, Nelson MA and Rosengren RJ: Mechanisms for the activity of heterocyclic cyclohexanone curcumin derivatives in estrogen receptor negative human breast cancer cell lines. Invest New Drugs. 29:87–97. 2011. View Article : Google Scholar
26.	Stuart EC and Rosengren RJ: The combination of raloxifene and epigallocatechin gallate suppresses growth and induces apoptosis in MDA-MB-231 cells. Life Sci. 82:943–948. 2008. View Article : Google Scholar : PubMed/NCBI
27.	Wang SC and Hung MC: HER2 overexpression and cancer targeting. Semin Oncol. 28:115–124. 2001. View Article : Google Scholar : PubMed/NCBI
28.	Hsieh WT, Huang KY, Lin HY and Chung JG: Physalis angulata induced G2/M phase arrest in human breast cancer cells. Food Chem Toxicol. 44:974–983. 2006. View Article : Google Scholar : PubMed/NCBI
29.	Hsu JD, Kao SH, Ou TT, Chen YJ, Li YJ and Wang CJ: Gallic acid induces G2/M phase arrest of breast cancer cell MCF-7 through stabilization of p27(Kip1) attributed to disruption of p27(Kip1)/Skp2 complex. J Agric Food Chem. 59:1996–2003. 2011. View Article : Google Scholar : PubMed/NCBI
30.	Santen RJ, Song RX, McPherson R, Kumar R, Adam L, Jeng MH and Yue W: The role of mitogen-activated protein (MAP) kinase in breast cancer. J Steroid Biochem Mol Biol. 80:239–256. 2002. View Article : Google Scholar : PubMed/NCBI
31.	Wada T and Penninger JM: Mitogen-activated protein kinases in apoptosis regulation. Oncogene. 23:2838–2849. 2004. View Article : Google Scholar : PubMed/NCBI
32.	Liu B, Han M, Sun RH, Wang JJ, Zhang YP, Zhang DQ and Wen JK: ABL-N-induced apoptosis in human breast cancer cells is partially mediated by c-Jun NH2-terminal kinase activation. Breast Cancer Res. 12:R92010. View Article : Google Scholar : PubMed/NCBI
33.	Kuo PL, Chen CY and Hsu YL: Isoobtusilactone a induces cell cycle arrest and apoptosis through reactive oxygen species/apoptosis signal-regulating kinase 1 signaling pathway in human breast cancer cells. Cancer Res. 67:7406–7420. 2007. View Article : Google Scholar
34.	Collett GP and Campbell FC: Curcumin induces c-jun N-terminal kinase-dependent apoptosis in HCT116 human colon cancer cells. Carcinogenesis. 25:2183–2189. 2004. View Article : Google Scholar : PubMed/NCBI
35.	Weir NM, Selvendiran K, Kutala VK, et al: Curcumin induces G2/M arrest and apoptosis in cisplatin-resistant human ovarian cancer cells by modulating Akt and p38 MAPK. Cancer Biol Ther. 6:178–184. 2007. View Article : Google Scholar : PubMed/NCBI
36.	Eto I: Nutritional and chemopreventive anti-cancer agents up-regulate expression of p27Kip1, a cyclin-dependent kinase inhibitor, in mouse JB6 epidermal and human MCF7, MDA-MB-321 and AU565 breast cancer cells. Cancer Cell Int. 6:202006. View Article : Google Scholar : PubMed/NCBI
37.	Dillon RL, White DE and Muller WJ: The phosphatidyl inositol 3-kinase signaling network: implications for human breast cancer. Oncogene. 26:1338–1345. 2007. View Article : Google Scholar : PubMed/NCBI
38.	Vivanco I and Sawyers CL: The phosphatidylinositol 3-kinase AKT pathway in human cancer. Nat Rev Cancer. 2:489–501. 2002. View Article : Google Scholar : PubMed/NCBI
39.	Burow ME, Weldon CB, Melnik LI, Duong BN, Collins-Burow BM, Beckman BS and McLachlan JA: PI3-K/AKT regulation of NF-kappaB signaling events in suppression of TNF-induced apoptosis. Biochem Biophys Res Commun. 271:342–345. 2000. View Article : Google Scholar : PubMed/NCBI
40.	Gong L, Li Y, Nedeljkovic-Kurepa A and Sarkar FH: Inactivation of NF-kappaB by genistein is mediated via Akt signaling pathway in breast cancer cells. Oncogene. 22:4702–4709. 2003. View Article : Google Scholar : PubMed/NCBI
41.	Barkett M and Gilmore TD: Control of apoptosis by Rel/NF-kappaB transcription factors. Oncogene. 18:6910–6924. 1999. View Article : Google Scholar : PubMed/NCBI
42.	Lauder A, Castellanos A and Weston K: c-Myb transcription is activated by protein kinase B (PKB) following interleukin 2 stimulation of T cells and is required for PKB-mediated protection from apoptosis. Mol Cell Biol. 21:5797–5805. 2001. View Article : Google Scholar : PubMed/NCBI
43.	Shehzad A, Wahid F and Lee YS: Curcumin in cancer chemoprevention: molecular targets, pharmacokinetics, bioavailability, and clinical trials. Arch Pharm. 343:489–499. 2010. View Article : Google Scholar : PubMed/NCBI
44.	Dhandapani KM, Mahesh VB and Brann DW: Curcumin suppresses growth and chemoresistance of human glioblastoma cells via AP-1 and NFkappaB transcription factors. J Neurochem. 102:522–538. 2007. View Article : Google Scholar : PubMed/NCBI
45.	Kasinski AL, Du Y, Thomas SL, et al: Inhibition of IkappaB kinase-nuclear factor-kappaB signaling pathway by 3,5-bis(2-flurobenzylidene)piperidin-4-one (EF24), a novel monoketone analog of curcumin. Mol Pharmacol. 74:654–661. 2008. View Article : Google Scholar : PubMed/NCBI
46.	Al-Hujaily EM, Mohamed AG, Al-Sharif I, et al: PAC, a novel curcumin analogue, has anti-breast cancer properties with higher efficiency on Er-negative cells. Breast Cancer Res Treat. 128:97–107. 2010. View Article : Google Scholar : PubMed/NCBI
47.	Hutzen B, Friedman L, Sobo M, et al: Curcumin analogue GO-Y030 inhibits STAT3 activity and cell growth in breast and pancreatic carcinomas. Int J Oncol. 35:867–872. 2009.PubMed/NCBI
48.	Kumar AP, Garcia GE, Ghosh R, Rajnarayanan RV, Alworth WL and Slaga TJ: 4-Hydroxy-3-methoxybenzoic acid methyl ester: a curcumin derivative targets Akt/NF kappa B cell survival signaling pathway: potential for prostate cancer management. Neoplasia. 5:255–266. 2003. View Article : Google Scholar : PubMed/NCBI
49.	Lin L, Hutzen B, Ball S, et al: New curcumin analogues exhibit enhanced growth-suppressive activity and inhibit AKT and signal transducer and activator of transcription 3 phosphorylation in breast and prostate cancer cells. Cancer Sci. 100:1719–1727. 2009. View Article : Google Scholar : PubMed/NCBI
50.	Anand P, Kunnumakkara AB, Newman RA and Aggarwal BB: Bioavailability of curcumin: problems and promises. Mol Pharm. 4:807–818. 2007. View Article : Google Scholar : PubMed/NCBI