Inactivation of AKT/NF‑κB signaling by eurycomalactone decreases human NSCLC cell viability and improves the chemosensitivity to cisplatin

Dukaew,Nahathai; Chairatvit,Kongthawat; Pitchakarn,Pornsiri; Imsumran,Arisa; Karinchai,Jirarat; Tuntiwechapikul,Wirote; Wongnoppavich,Ariyaphong

doi:10.3892/or.2020.7710

Inactivation of AKT/NF‑κB signaling by eurycomalactone decreases human NSCLC cell viability and improves the chemosensitivity to cisplatin

Authors:
- Nahathai Dukaew
- Kongthawat Chairatvit
- Pornsiri Pitchakarn
- Arisa Imsumran
- Jirarat Karinchai
- Wirote Tuntiwechapikul
- Ariyaphong Wongnoppavich
View Affiliations

Affiliations: Graduate/PhD Degree Program in Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand, Department of Oral Biology, Faculty of Dentistry, Mahidol University, Bangkok 10400, Thailand, Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand
Published online on: July 31, 2020 https://doi.org/10.3892/or.2020.7710
Pages: 1441-1454
Copyright: © Dukaew et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

The high activation of protein kinase B (AKT)/nuclear factor‑κB (NF‑κB) signaling has often been associated with the induction of non‑small cell lung cancer (NSCLC) cell survival and resistance to cisplatin, which is one of the most widely used chemotherapeutic drugs in the treatment of NSCLC. The inhibition of AKT/NF‑κB can potentially be used as a molecular target for cancer therapy. Eurycomalactone (ECL), a quassinoid from Eurycoma longifolia Jack, has previously been revealed to exhibit strong cytotoxic activity against the human NSCLC A549 cell line, and can inhibit NF‑κB activity in TNF‑α‑activated 293 cells stably transfected with an NF‑κB luciferase reporter. The present study was the first to investigate whether ECL inhibits the activation of AKT/NF‑κB signaling, induces apoptosis and enhances chemosensitivity to cisplatin in human NSCLC cells. The anticancer activity of ECL was evaluated in two NSCLC cell lines, A549 and Calu‑1. ECL decreased the viability and colony formation ability of both cell lines by inducing cell cycle arrest and apoptosis through the activation of pro‑apoptotic caspase‑3 and poly (ADP‑ribose) polymerase, as well as the reduction of anti‑apoptotic proteins Bcl‑xL and survivin. In addition, ECL treatment suppressed the levels of AKT (phospho Ser473) and NF‑κB (phospho Ser536). Notably, ECL significantly enhanced cisplatin sensitivity in both assessed NSCLC cell lines. The combination treatment of cisplatin and ECL promoted cell apoptosis more effectively than cisplatin alone, as revealed by the increased cleaved caspase‑3, but decreased Bcl‑xL and survivin levels. Exposure to cisplatin alone induced the levels of phosphorylated‑AKT and phosphorylated‑NF‑κB, whereas co‑treatment with ECL inhibited the cisplatin‑induced phosphorylation of AKT and NF‑κB, leading to an increased sensitization effect on cisplatin‑induced apoptosis. In conclusion, ECL exhibited an anticancer effect and sensitized NSCLC cells to cisplatin through the inactivation of AKT/NF‑κB signaling. This finding provides a rationale for the combined use of chemotherapy drugs with ECL to improve their efficacy in NSCLC treatment.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2018. CA Cancer J Clin. 68:7–30. 2018. View Article : Google Scholar : PubMed/NCBI
2	Liloglou T, Bediaga NG, Brown BR, Field JK and Davies MP: Epigenetic biomarkers in lung cancer. Cancer Lett. 342:200–212. 2014. View Article : Google Scholar : PubMed/NCBI
3	D'Addario G, Pintilie M, Leighl NB, Feld R, Cerny T and Shepherd FA: Platinum-based versus non-platinum-based chemotherapy in advanced non-small-cell lung cancer: A meta-analysis of the published literature. J Clin Oncol. 23:2926–2936. 2005. View Article : Google Scholar : PubMed/NCBI
4	Pei X, Xiao J, Wei G, Zhang Y, Lin F, Xiong Z, Lu L, Wang X, Pang G, Jiang Y and Jiang L: Oenothein B inhibits human non-small cell lung cancer A549cell proliferation by ROS-mediated PI3K/Akt/NF-κB signaling pathway. Chem Biol Interact. 298:112–120. 2019. View Article : Google Scholar : PubMed/NCBI
5	Hafsi S, Pezzino FM, Candido S, Ligresti G, Spandidos DA, Soua Z, McCubrey JA, Travali S and Libra M: Gene alterations in the PI3K/PTEN/AKT pathway as a mechanism of drug-resistance (review). Int J Oncol. 40:639–644. 2012.PubMed/NCBI
6	Hakan Kucuksayan H, Sakir Akgun S and Akca H: Pl3K/Akt/NF-κB signalling pathway on NSCLC invasion. Med Chem. 6:42016.
7	Naugler WE and Karin M: NF-kappaB and cancer-identifying targets and mechanisms. Curr Opin Genet Dev. 18:19–26. 2008. View Article : Google Scholar : PubMed/NCBI
8	Chen W, Liu X, Yuan S and Qiao T: HSPA12B overexpression induces cisplatin resistance in non-small-cell lung cancer by regulating the PI3K/Akt/NF-κB signaling pathway. Oncol Lett. 15:3883–3889. 2018.PubMed/NCBI
9	Zou W, Ma X, Hua W, Chen B and Cai G: Caveolin-1 mediates chemoresistance in cisplatin-resistant ovarian cancer cells by targeting apoptosis through the Notch-1/Akt/NF-κB pathway. Oncol Rep. 34:3256–3263. 2015. View Article : Google Scholar : PubMed/NCBI
10	Godwin P, Baird AM, Heavey S, Barr MP, O'Byrne KJ and Gately K: Targeting nuclear factor-kappa B to overcome resistance to chemotherapy. Front Oncol. 3:1202013. View Article : Google Scholar : PubMed/NCBI
11	Albanell J, Chattopadhyay S, Tapia M, Rovira A, Belda-Iniesta C, Manguán-García JDC, Machado R, Gonzalez Barón M and Perona R: JNK/MKP-1 and PI3K/NF-κB: Critical pathways controlling cellular response towards cisplatin in non-small cell lung cancer (NSCLC). J Clin Oncol. 23:2033. 2005. View Article : Google Scholar
12	Nobili S, Lippi D, Witort E, Donnini M, Bausi L, Mini E and Capaccioli S: Natural compounds for cancer treatment and prevention. Pharmacol Res. 59:365–378. 2009. View Article : Google Scholar : PubMed/NCBI
13	Li Y, Ahmed F, Ali S, Philip PA, Kucuk O and Sarkar FH: Inactivation of nuclear factor kappaB by soy isoflavone genistein contributes to increased apoptosis induced by chemotherapeutic agents in human cancer cells. Cancer Res. 65:6934–6942. 2005. View Article : Google Scholar : PubMed/NCBI
14	Liu D, Yan L, Wang L, Tai W, Wang W and Yang C: Genistein enhances the effect of cisplatin on the inhibition of non-small cell lung cancer A549 cell growth in vitro and in vivo. Oncol Lett. 8:2806–2810. 2014. View Article : Google Scholar : PubMed/NCBI
15	Yu M, Qi B, Wu X, Xu J and Liu X: Baicalein increases cisplatin sensitivity of A549 lung adenocarcinoma cells via PI3K/Akt/NF-κB pathway. Biomed Pharmacother. 90:677–685. 2017. View Article : Google Scholar : PubMed/NCBI
16	Ahmmed B, Khan MN, Nisar MA, Kampo S, Zheng Q, Li Y and Yan Q: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-kappaB signaling pathway. Int J Oncol. 54:431–442. 2019.PubMed/NCBI
17	Miyake K, Tezuka Y, Awale S, Li F and Kadota S: Quassinoids from Eurycoma longifolia. J Nat Prod. 72:2135–2140. 2009. View Article : Google Scholar : PubMed/NCBI
18	Rehman SU, Choe K and Yoo HH: Review on a traditional herbal medicine, Eurycoma longifolia Jack (tongkat ali): Its traditional uses, chemistry, evidence-based pharmacology and toxicology. Molecules. 21:3312016. View Article : Google Scholar : PubMed/NCBI
19	Kuo PC, Damu AG, Lee KH and Wu TS: Cytotoxic and antimalarial constituents from the roots of Eurycoma longifolia. Bioorg Med Chem. 12:537–544. 2004. View Article : Google Scholar : PubMed/NCBI
20	Miyake K, Li F, Tezuka Y, Awale S and Kadota S: Cytotoxic activity of quassinoids from Eurycoma longifolia. Nat Prod Commun. 5:1009–1012. 2010.PubMed/NCBI
21	Dukaew N, Konishi T, Chairatvit K, Autsavapromporn N, Soonthornchareonnon N and Wongnoppavich A: Enhancement of radiosensitivity by eurycomalactone in human NSCLC cells through G2/M cell cycle arrest and delayed DNA double-strand break repair. Oncol Res. 28:161–175. 2020. View Article : Google Scholar : PubMed/NCBI
22	Tran TV, Malainer C, Schwaiger S, Atanasov AG, Heiss EH, Dirsch VM and Stuppner H: NF-κB inhibitors from Eurycoma longifolia. J Nat Prod. 77:483–488. 2014. View Article : Google Scholar : PubMed/NCBI
23	Malainer C, Schachner D, Sangiovanni E, Atanasov AG, Schwaiger S, Stuppner H, Heiss EH and Dirsch VM: Eurycomalactone inhibits expression of endothelial adhesion molecules at a post-transcriptional level. J Nat Prod. 80:3186–3193. 2017. View Article : Google Scholar : PubMed/NCBI
24	Phannasorn W, Khanaree C, Wongnoppavich A and Chewonarin T: The effect of purple rice (Oryza sativa L. indica) extract on the inflammatory response in a colon cancer cell line and dextran sulfate-induced tumor promotion in the rat colon. Mol Cell Toxicol. 13:433–442. 2017. View Article : Google Scholar
25	Peters GJ, van der Wilt CL, van Moorsel CJ, Kroep JR, Bergman AM and Ackland SP: Basis for effective combination cancer chemotherapy with antimetabolites. Pharmacol Ther. 87:227–253. 2000. View Article : Google Scholar : PubMed/NCBI
26	Chou TC: Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 70:440–446. 2010. View Article : Google Scholar : PubMed/NCBI
27	Wongnoppavich A, Dukaew N, Choonate S and Chairatvit K: Upregulation of maspin expression in human cervical carcinoma cells by transforming growth factor beta1 through the convergence of Smad and non-Smad signaling pathways. Oncol Lett. 13:3646–3652. 2017. View Article : Google Scholar : PubMed/NCBI
28	Kaltschmidt C, Banz-Jansen C, Benhidjeb T, Beshay M, Förster C, Greiner J, Hamelmann E, Jorch N, Mertzlufft F, Pfitzenmaier J, et al: A role for NF-κB in organ specific cancer and cancer stem cells. Cancers (Basel). 11:6552019. View Article : Google Scholar
29	Huang WC and Hung MC: Induction of Akt activity by chemotherapy confers acquired resistance. J Formos Med Assoc. 108:180–194. 2009. View Article : Google Scholar : PubMed/NCBI
30	Liao Y and Hung MC: Physiological regulation of Akt activity and stability. Am J Transl Res. 2:19–42. 2010.PubMed/NCBI
31	Hu J, Nakano H, Sakurai H and Colburn NH: Insufficient p65 phosphorylation at S536 specifically contributes to the lack of NF-kappaB activation and transformation in resistant JB6 cells. Carcinogenesis. 25:1991–2003. 2004. View Article : Google Scholar : PubMed/NCBI
32	Barta JA, Powell CA and Wisnivesky JP: Global epidemiology of lung cancer. Ann Glob Health. 85:82019. View Article : Google Scholar : PubMed/NCBI
33	Ijaz S, Akhtar N, Khan MS, Hameed A, Irfan M, Arshad MA, Ali S and Asrar M: Plant derived anticancer agents: A green approach towards skin cancers. Biomed Pharmacother. 103:1643–1651. 2018. View Article : Google Scholar : PubMed/NCBI
34	Wong PF, Cheong WF, Shu MH, Teh CH, Chan KL and AbuBakar S: Eurycomanone suppresses expression of lung cancer cell tumor markers, prohibitin, annexin 1 and endoplasmic reticulum protein 28. Phytomedicine. 19:138–144. 2012. View Article : Google Scholar : PubMed/NCBI
35	Zakaria Y, Rahmat A, Pihie AH, Abdullah NR and Houghton PJ: Eurycomanone induce apoptosis in HepG2 cells via up- regulation of p53. Cancer Cell Int. 9:162009. View Article : Google Scholar : PubMed/NCBI
36	Hajjouli S, Chateauvieux S, Teiten MH, Orlikova B, Schumacher M, Dicato M, Choo CY and Diederich M: Eurycomanone and eurycomanol from Eurycoma longifolia Jack as regulators of signaling pathways involved in proliferation, cell death and inflammation. Molecules. 19:14649–14666. 2014. View Article : Google Scholar : PubMed/NCBI
37	Tokunaga E, Oki E, Egashira A, Sadanaga N, Morita M, Kakeji Y and Maehara Y: Deregulation of the Akt pathway in human cancer. Curr Cancer Drug Targets. 8:27–36. 2008. View Article : Google Scholar : PubMed/NCBI
38	Bai D, Ueno L and Vogt PK: Akt-mediated regulation of NFkappaB and the essentialness of NFkappaB for the oncogenicity of PI3K and Akt. Int J Cancer. 125:2863–2870. 2009. View Article : Google Scholar : PubMed/NCBI
39	Oeckinghaus A and Ghosh S: The NF-kappaB family of transcription factors and its regulation. Cold Spring Harb Perspect Biol. 1:a0000342009. View Article : Google Scholar : PubMed/NCBI
40	Chen W, Li Z, Bai L and Lin Y: NF-kappaB in lung cancer, a carcinogenesis mediator and a prevention and therapy target. Front Biosci (Landmark Ed). 16:1172–1185. 2011. View Article : Google Scholar : PubMed/NCBI
41	Umemura S, Mimaki S, Makinoshima H, Tada S, Ishii G, Ohmatsu H, Niho S, Yoh K, Matsumoto S, Takahashi A, et al: Therapeutic priority of the PI3K/AKT/mTOR pathway in small cell lung cancers as revealed by a comprehensive genomic analysis. J Thorac Oncol. 9:1324–1331. 2014. View Article : Google Scholar : PubMed/NCBI
42	Garcia-Echeverria C and Sellers WR: Drug discovery approaches targeting the PI3K/Akt pathway in cancer. Oncogene. 27:5511–5526. 2008. View Article : Google Scholar : PubMed/NCBI
43	Gu JJ, Qiao KS, Sun P, Chen P and Li Q: Study of EGCG induced apoptosis in lung cancer cells by inhibiting PI3K/Akt signaling pathway. Eur Rev Med Pharmacol Sci. 22:4557–4563. 2018.PubMed/NCBI
44	Li JY, Yu J, Du XS, Zhang HM, Wang B, Guo H, Bai J, Wang JH, Liu A and Wang YL: Safflower polysaccharide induces NSCLC cell apoptosis by inhibition of the Akt pathway. Oncol Rep. 36:147–154. 2016. View Article : Google Scholar : PubMed/NCBI
45	Kwon HJ, Choi GE, Ryu S, Kwon SJ, Kim SC, Booth C, Nichols KE and Kim HS: Stepwise phosphorylation of p65 promotes NF-κB activation and NK cell responses during target cell recognition. Nat Commun. 7:116862016. View Article : Google Scholar : PubMed/NCBI
46	Zheng J, Son DJ, Gu SM, Woo JR, Ham YW, Lee HP, Kim WJ, Jung JK and Hong JT: Piperlongumine inhibits lung tumor growth via inhibition of nuclear factor kappa B signaling pathway. Sci Rep. 6:263572016. View Article : Google Scholar : PubMed/NCBI
47	Khan KH, Yap TA, Yan L and Cunningham D: Targeting the PI3K-AKT-mTOR signaling network in cancer. Chin J Cancer. 32:253–265. 2013. View Article : Google Scholar : PubMed/NCBI
48	Shukla S, Shankar E, Fu P, MacLennan GT and Gupta S: Suppression of NF-κB and NF-κB-regulated gene expression by apigenin through IκBα and IKK pathway in TRAMP mice. PLoS One. 10:e01387102015. View Article : Google Scholar : PubMed/NCBI
49	Cheung CH, Huang CC, Tsai FY, Lee JY, Cheng SM, Chang YC, Huang YC, Chen SH and Chang JY: Survivin-biology and potential as a therapeutic target in oncology. Onco Targets Ther. 6:1453–1462. 2013. View Article : Google Scholar : PubMed/NCBI
50	Rinaldi M, Cauchi C and Gridelli C: First line chemotherapy in advanced or metastatic NSCLC. Ann Oncol. 17 (Suppl 5):v64–v67. 2006. View Article : Google Scholar : PubMed/NCBI
51	Galluzzi L, Senovilla L, Vitale I, Michels J, Martins I, Kepp O, Castedo M and Kroemer G: Molecular mechanisms of cisplatin resistance. Oncogene. 31:1869–1883. 2012. View Article : Google Scholar : PubMed/NCBI
52	Köberle B, Tomicic MT, Usanova S and Kaina B: Cisplatin resistance: Preclinical findings and clinical implications. Biochim Biophys Acta. 1806:172–182. 2010.PubMed/NCBI
53	Baby J, Pickering BF, Vashisht Gopal YN and Van Dyke MW: Constitutive and inducible nuclear factor-kappaB in immortalized normal human bronchial epithelial and non-small cell lung cancer cell lines. Cancer Lett. 255:85–94. 2007. View Article : Google Scholar : PubMed/NCBI
54	Chen B, Shen Z, Wu D, Xie X, Xu X, Lv L, Dai H, Chen J and Gan X: Glutathione peroxidase 1 promotes NSCLC resistance to cisplatin via ROS-induced activation of PI3K/AKT pathway. Biomed Res Int. 2019:76405472019.PubMed/NCBI
55	Li Z, Yang Z, Lapidus RG, Liu X, Cullen KJ and Dan HC: IKK phosphorylation of NF-κB at serine 536 contributes to acquired cisplatin resistance in head and neck squamous cell cancer. Am J Cancer Res. 5:3098–3110. 2015.PubMed/NCBI