Dammarane-type triterpene ginsenoside-Rg18 inhibits human non-small cell lung cancer A549 cell proliferation via G1 phase arrest

Leem,Dong‑Gyu; Shin,Ji‑Sun; Kim,Kyung‑Tack; Choi,Sang  Yoon; Lee,Myung‑Hee; Lee,Kyung‑Tae

doi:10.3892/ol.2018.8057

Dammarane-type triterpene ginsenoside-Rg18 inhibits human non-small cell lung cancer A549 cell proliferation via G1 phase arrest

Authors:
- Dong‑Gyu Leem
- Ji‑Sun Shin
- Kyung‑Tack Kim
- Sang Yoon Choi
- Myung‑Hee Lee
- Kyung‑Tae Lee
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Affiliations: Department of Pharmaceutical Biochemistry, College of Pharmacy, Kyung Hee University, Seoul 02447, Republic of Korea, Traditional Food Research Center, Korea Food Research Institute, Seongnam 13539, Republic of Korea
Published online on: February 15, 2018 https://doi.org/10.3892/ol.2018.8057
Pages: 6043-6049

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Abstract

A previous study reported that a novel dammarane-type triterpene saponin, ginsenoside-Rg18, derived from the root of Panax ginseng, displayed hydroxyl radical scavenging, anti‑bacterial and cytotoxic activities. However, the underlying molecular mechanisms of its anti‑proliferative effect on non‑small cell lung cancer (NSCLC) A549 cells remains unclear. In the present study, it was determined that Rg18 inhibited the proliferation of A549 cells with a half‑maximal inhibitory concentration of 150 µM. Flow cytometry analysis indicated that cell cycle progression was blocked by Rg18 at G1 phase in A549 cells, which was accompanied by downregulation of cyclin‑dependent kinase 2 (CDK2), CDK4, CDK6, cyclin D1, cyclin D2, cyclin E and phosphorylated retinoblastoma protein expression at the protein level. In addition, the CDK inhibitors (CDKNs), CDKN1A and CDKN1B, were upregulated following Rg18 treatment. Furthermore, Rg18 treatment resulted in the intracellular accumulation of reactive oxygen species (ROS), and a dose‑dependent inhibition of p38 mitogen activated protein kinase (p38), c‑Jun N‑terminal kinase (JNK) and nuclear factor‑κB (NF‑κB)/p65 phosphorylation. Taken together, Rg18‑mediated G1 phase arrest was closely associated with the generation of intracellular ROS, and p38, JNK and NF‑κB/p65 inhibition in A549 human NSCLC cells.

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1	Torre LA, Siegel RL and Jemal A: Lung cancer statistics. Adv Exp Med Biol. 893:1–19. 2016. View Article : Google Scholar : PubMed/NCBI
2	Molina JR, Yang P, Cassivi SD, Schild SE and Adjei AA: Non-small cell lung cancer: Epidemiology, risk factors, treatment, and survivorship. Mayo Clin Proc. 83:pp. 584–594. 2008; View Article : Google Scholar : PubMed/NCBI
3	Huang CY, Ju DT, Chang CF, Muralidhar Reddy P and Velmurugan BK: A review on the effects of current chemotherapy drugs and natural agents in treating non-small cell lung cancer. Biomedicine (Taipei). 7:232017. View Article : Google Scholar : PubMed/NCBI
4	Chen HS, Liu Y, Lin LQ, Zhao JL, Zhang CP, Jin JC, Wang L, Bai MH, Wang YC, Liu M and Shen BZ: Anti-proliferative effect of an extract of the root of Polygonum multiflorum Thunb. On MCF-7 human breast cancer cells and the possible mechanisms. Mol Med Rep. 4:1313–1319. 2011.PubMed/NCBI
5	Dickson MA and Schwartz GK: Development of cell-cycle inhibitors for cancer therapy. Curr Oncol. 16:36–43. 2009.PubMed/NCBI
6	Giacinti C and Giordano A: RB and cell cycle progression. Oncogene. 25:5220–5227. 2006. View Article : Google Scholar : PubMed/NCBI
7	Foster DA, Yellen P, Xu L and Saqcena M: Regulation of G1 cell cycle progression: Distinguishing the restriction point from a nutrient-sensing cell growth checkpoint(s). Genes Cancer. 1:1124–1131. 2010. View Article : Google Scholar : PubMed/NCBI
8	Nita M and Grzybowski A: The role of the reactive oxygen species and oxidative stress in the pathomechanism of the age-related ocular diseases and other pathologies of the anterior and posterior eye segments in adults. Oxid Med Cell Longev. 2016:31647342016. View Article : Google Scholar : PubMed/NCBI
9	Ray PD, Huang BW and Tsuji Y: Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal. 24:981–990. 2012. View Article : Google Scholar : PubMed/NCBI
10	Wang Z, Zhao X and Gong X: Costunolide induces lung adenocarcinoma cell line A549 cells apoptosis through ROS (reactive oxygen species)-mediated endoplasmic reticulum stress. Cell Biol Int. 40:289–297. 2016. View Article : Google Scholar : PubMed/NCBI
11	Xiao W, Jiang Y, Men Q, Yuan L, Huang Z, Liu T, Li W and Liu X: Tetrandrine induces G1/S cell cycle arrest through the ROS/Akt pathway in EOMA cells and inhibits angiogenesis in vivo. Int J Oncol. 46:360–368. 2015. View Article : Google Scholar : PubMed/NCBI
12	Qi B, Zhang L, Zhang Z, Ouyang J and Huang H: Effects of ginsenosides-Rb1 on exercise-induced oxidative stress in forced swimming mice. Pharmacogn Mag. 10:458–463. 2014. View Article : Google Scholar : PubMed/NCBI
13	Li J, Zhong W, Wang W, Hu S, Yuan J, Zhang B, Hu T and Song G: Ginsenoside metabolite compound K promotes recovery of dextran sulfate sodium-induced colitis and inhibits inflammatory responses by suppressing NF-κB activation. PLoS One. 9:e878102014. View Article : Google Scholar : PubMed/NCBI
14	Chung KS, Cho SH, Shin JS, Kim DH, Choi JH, Choi SY, Rhee YK, Hong HD and Lee KT: Ginsenoside Rh2 induces cell cycle arrest and differentiation in human leukemia cells by upregulating TGF-β expression. Carcinogenesis. 34:331–340. 2013. View Article : Google Scholar : PubMed/NCBI
15	Lee DG, Lee AY, Kim KT, Cho EJ and Lee S: Novel dammarane-type triterpene saponins from Panax ginseng root. Chem Pharm Bull (Tokyo). 63:927–934. 2015. View Article : Google Scholar : PubMed/NCBI
16	Plumb JA, Milroy R and Kaye SB: Effects of the pH dependence of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide-formazan absorption on chemosensitivity determined by a novel tetrazolium-based assay. Cancer Res. 49:4435–4440. 1989.PubMed/NCBI
17	Choi JH and Lee KT: Costunolide-induced apoptosis in human leukemia cells: Involvement of c-jun N-terminal kinase activation. Biol Pharm Bull. 32:1803–1808. 2009. View Article : Google Scholar : PubMed/NCBI
18	Park EY, Kim JI, Leem DG, Shin JS, Kim KT, Choi SY, Lee MH, Choi JH, Lee YS and Lee KT: Resveratrol analogue (E)-8-acetoxy-2-[2-(3,4-diacetoxyphenyl)ethenyl]-quinazoline induces apoptosis via Fas-mediated pathway in HL-60 human leukemia cells. Oncol Rep. 36:3577–3587. 2016. View Article : Google Scholar : PubMed/NCBI
19	Taya Y: RB kinases and RB-binding proteins: New points of view. Trends Biochem Sci. 22:14–17. 1997. View Article : Google Scholar : PubMed/NCBI
20	Verbon EH, Post JA and Boonstra J: The influence of reactive oxygen species on cell cycle progression in mammalian cells. Gene. 511:1–6. 2012. View Article : Google Scholar : PubMed/NCBI
21	Zhang W and Liu HT: MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res. 12:9–18. 2002. View Article : Google Scholar : PubMed/NCBI
22	Joyce D, Albanese C, Steer J, Fu M, Bouzahzah B and Pestell RG: NF-kappaB and cell-cycle regulation: The cyclin connection. Cytokine Growth Factor Rev. 12:73–90. 2001. View Article : Google Scholar : PubMed/NCBI
23	Abe H, Arichi S, Hayashi T and Odashima S: Ultrastructural studies of Morris hepatoma cells reversely transformed by ginsenosides. Experientia. 35:1647–1649. 1979. View Article : Google Scholar : PubMed/NCBI
24	Kim HS, Lee EH, Ko SR, Choi KJ, Park JH and Im DS: Effects of ginsenosides Rg3 and Rh2 on the proliferation of prostate cancer cells. Arch Pharm Res. 27:429–435. 2004. View Article : Google Scholar : PubMed/NCBI
25	Cheng CC, Yang SM, Huang CY, Chen JC, Chang WM and Hsu SL: Molecular mechanisms of ginsenoside Rh2-mediated G1 growth arrest and apoptosis in human lung adenocarcinoma A549 cells. Cancer Chemother Pharmacol. 55:531–540. 2005. View Article : Google Scholar : PubMed/NCBI
26	Hwang HC and Clurman BE: Cyclin E in normal and neoplastic cell cycles. Oncogene. 24:2776–2786. 2005. View Article : Google Scholar : PubMed/NCBI
27	Connell-Crowley L, Harper JW and Goodrich DW: Cyclin D1/Cdk4 regulates retinoblastoma protein-mediated cell cycle arrest by site-specific phosphorylation. Mol Biol Cell. 8:287–301. 1997. View Article : Google Scholar : PubMed/NCBI
28	Schmidt M, Lu Y, Parant JM, Lozano G, Bacher G, Beckers T and Fan Z: Differential roles of p21(Waf1) and p27(Kip1) in modulating chemosensitivity and their possible application in drug discovery studies. Mol Pharmacol. 60:900–906. 2001. View Article : Google Scholar : PubMed/NCBI
29	Lee KW, Kim HJ, Lee YS, Park HJ, Choi JW, Ha J and Lee KT: Acteoside inhibits human promyelocytic HL-60 leukemia cell proliferation via inducing cell cycle arrest at G0/G1 phase and differentiation into monocyte. Carcinogenesis. 28:1928–1936. 2007. View Article : Google Scholar : PubMed/NCBI
30	Wang R, Zhang Q, Peng X, Zhou C, Zhong Y, Chen X, Qiu Y, Jin M, Gong M and Kong D: Stellettin B induces G1 arrest, apoptosis and autophagy in human non-small cell lung cancer A549 cells via blocking PI3K/Akt/mTOR pathway. Sci Rep. 6:270712016. View Article : Google Scholar : PubMed/NCBI
31	Chang MS, Lee SG and Rho HM: Transcriptional activation of Cu/Zn superoxide dismutase and catalase genes by panaxadiol ginsenosides extracted from Panax ginseng. Phytother Res. 13:641–644. 1999. View Article : Google Scholar : PubMed/NCBI
32	Dhillon AS, Hagan S, Rath O and Kolch W: MAP kinase signalling pathways in cancer. Oncogene. 26:3279–3290. 2007. View Article : Google Scholar : PubMed/NCBI
33	Roberts PJ and Der CJ: Targeting the Raf-MEK-ERK mitogen-activated protein kinase cascade for the treatment of cancer. Oncogene. 26:3291–3310. 2007. View Article : Google Scholar : PubMed/NCBI
34	Wagner EF and Nebreda AR: Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer. 9:537–549. 2009. View Article : Google Scholar : PubMed/NCBI
35	Gao JL, Lv GY, He BC, Zhang BQ, Zhang H, Wang N, Wang CZ, Du W, Yuan CS and He TC: Ginseng saponin metabolite 20(S)-protopanaxadiol inhibits tumor growth by targeting multiple cancer signaling pathways. Oncol Rep. 30:292–298. 2013. View Article : Google Scholar : PubMed/NCBI
36	Jung SH, Woo MS, Kim SY, Kim WK, Hyun JW, Kim EJ, Kim DH and Kim HS: Ginseng saponin metabolite suppresses phorbol ester-induced matrix metalloproteinase-9 expression through inhibition of activator protein-1 and mitogen-activated protein kinase signaling pathways in human astroglioma cells. Int J Cancer. 118:490–497. 2006. View Article : Google Scholar : PubMed/NCBI
37	An JJ, Shi KJ, Wei W, Hua FY, Ci YL, Jiang Q, Li F, Wu P, Hui KY, Yang Y and Xu CM: The ROS/JNK/ATF2 pathway mediates selenite-induced leukemia NB4 cell cycle arrest and apoptosis in vitro and in vivo. Cell Death Dis. 4:e9732013. View Article : Google Scholar : PubMed/NCBI
38	Kaltschmidt B, Kaltschmidt C, Hehner SP, Dröge W and Schmitz ML: Repression of NF-kappaB impairs HeLa cell proliferation by functional interference with cell cycle checkpoint regulators. Oncogene. 18:3213–3225. 1999. View Article : Google Scholar : PubMed/NCBI
39	Rajitha B, Belalcazar A, Nagaraju GP, Shaib WL, Snyder JP, Shoji M, Pattnaik S, Alam A and El-Rayes BF: Inhibition of NF-κB translocation by curcumin analogs induces G0/G1 arrest and downregulates thymidylate synthase in colorectal cancer. Cancer Lett. 373:227–233. 2016. View Article : Google Scholar : PubMed/NCBI
40	Tran KQ, Tin AS and Firestone GL: Artemisinin triggers a G1 cell cycle arrest of human Ishikawa endometrial cancer cells and inhibits cyclin-dependent kinase-4 promoter activity and expression by disrupting nuclear factor-κB transcriptional signaling. Anticancer Drugs. 25:270–281. 2014. View Article : Google Scholar : PubMed/NCBI
41	Kim SM, Lee SY, Cho JS, Son SM, Choi SS, Yun YP, Yoo HS, Yoon DY, Oh KW, Han SB and Hong JT: Combination of ginsenoside Rg3 with docetaxel enhances the susceptibility of prostate cancer cells via inhibition of NF-kappaB. Eur J Pharmacol. 631:1–9. 2010. View Article : Google Scholar : PubMed/NCBI
42	Kim ND, Pokharel YR and Kang KW: Ginsenoside Rd enhances glutathione levels in H4IIE cells via NF-kappaB-dependent gamma-glutamylcysteine ligase induction. Pharmazie. 62:933–936. 2007.PubMed/NCBI