Abstract
In this study, we investigated the anti-cancer effects of ginsenoside Rg2 (G-Rg2) and its underlying signaling pathways in breast cancer (BC) cells. G-Rg2 significantly induced cytotoxicity and reactive oxygen species (ROS) production in MCF-7 cells among various types of BC cells including HCC1428, T47D, and BT-549. G-Rg2 significantly inhibited protein and mRNA expression of cell cycle G1-S phase regulators, including p-Rb, cyclin D1, CDK4, and CDK6, whereas it enhanced the protein and mRNA expression of cell cycle arrest and apoptotic molecules including cleaved PARP, p21, p27, p53 and Bak through ROS production. These effects were abrogated by the antioxidant N-acetyl-I-cysteine, or NADPH oxidase inhibitors, such as diphenyleneiodonium chloride and apocynin. Interestingly, G-Rg2 induced mitochondrial damage by reducing the membrane potential. G-Rg2 further activated the ROS-sensor protein, AMPK and downstream targets of AMPK activation, including PGC-1α, FOXO1, and IDH2, and downregulated mTOR activation and antioxidant response element-driven luciferase activity. Together, our data demonstrate that G-Rg2 mediates anti-cancer effects by activating cell cycle arrest and signaling pathways related to mitochondrial damage-induced ROS production and apoptosis.
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References
Ahn J, Kim H, Yang KM (2020) Omega-hydroxyundec-9-enoic acid induction of breast cancer cells apoptosis through generation of mitochondrial ROS and phosphorylation of AMPK. Arch Pharm Res 43:735–743. https://doi.org/10.1007/s12272-020-01254-x
Azamjah N, Soltan-Zadeh Y, Zayeri F (2019) Global Trend of Breast Cancer Mortality Rate: A 25-Year Study. Asian Pac J Cancer Prev 20:2015–2020. https://doi.org/10.31557/APJCP.2019.20.7.2015
Bak MJ, Jeong WS, Kim KB (2014) Detoxifying effect of fermented black ginseng on H2O2-induced oxidative stress in HepG2 cells. Int J Mol Med 34:1516–1522. https://doi.org/10.3892/ijmm.2014.1972
Bost F, Kaminski L (2019) The metabolic modulator PGC-1alpha in cancer. Am J Cancer Res 9:198–211
Brandie N, Radde MM, Ivanova HX, Mai, Joshua K, Salabei BG, Hill, Carolyn M, Klinge (2015) Bioenergetic differences between MCF-7 and T47D breast cancer cells and their regulation by oestradiol and tamoxifen. Biochem J: 465(1):49–61. https://doi.org/10.1042/BJ20131608
Chaube B, Malvi P, Singh SV, Mohammad N, Viollet B, Bhat MK (2015) AMPK maintains energy homeostasis and survival in cancer cells via regulating p38/PGC-1alpha-mediated mitochondrial biogenesis. Cell Death Discov 1:15063. https://doi.org/10.1038/cddiscovery.2015.63
Cui J, Wang J, Zheng M, Gou D, Liu C, Zhou Y (2017) Ginsenoside Rg2 protects PC12 cells against beta-amyloid25-35-induced apoptosis via the phosphoinositide 3-kinase/Akt pathway. Chem Biol Interact 275:152–161. https://doi.org/10.1016/j.cbi.2017.07.021
Cui L, Bu W, Song J, Feng L, Xu T, Liu D, Ding W, Wang J, Li C, Ma B, Luo Y, Jiang Z, Wang C, Chen J, Hou J, Yan H, Yang L, Jia X (2018) Apoptosis induction by alantolactone in breast cancer MDA-MB-231 cells through reactive oxygen species-mediated mitochondrion-dependent pathway. Arch Pharm Res 41:299–313. https://doi.org/10.1007/s12272-017-0990-2
Dai X, Cheng H, Bai Z, Li J (2017) Breast cancer cell line classification and its relevance with breast tumor subtyping. J Cancer 8:3131–3141. https://doi.org/10.7150/jca.18457
Duarte FV, Amorim JA, Palmeira CM, Rolo AP (2015) Regulation of mitochondrial function and its impact in metabolic stress. Curr Med Chem 22:2468–2479. https://doi.org/10.2174/0929867322666150514095910
Gou D, Pei X, Wang J, Wang Y, Hu C, Song C, Cui S, Zhou Y (2020) Antiarrhythmic effects of ginsenoside Rg2 on calcium chloride-induced arrhythmias without oral toxicity. J Ginseng Res 44:717–724. https://doi.org/10.1016/j.jgr.2019.06.005
Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312. https://doi.org/10.1126/science.281.5381.1309
Gross DN, van den Heuvel AP, Birnbaum MJ (2008) The role of FoxO in the regulation of metabolism. Oncogene 27:2320–2336. https://doi.org/10.1038/onc.2008.25
Guo C, Sun L, Chen X, Zhang D (2013) Oxidative stress, mitochondrial damage and neurodegenerative diseases. Neural Regen Res 8:2003–2014. https://doi.org/10.3969/j.issn.1673-5374.2013.21.009
Huynh DTN, Baek N, Sim S, Myung CS, Heo KS (2020) Minor ginsenoside Rg2 and Rh1 attenuates LPS-induced acute liver and kidney damages via downregulating activation of TLR4-STAT1 and inflammatory cytokine production in macrophages. Int J Mol Sci. https://doi.org/10.3390/ijms21186656
Huynh DTN, Jin Y, Myung C-S, Heo K-S (2021) Ginsenoside Rh1 induces MCF-7 cell apoptosis and autophagic cell death through ROS-mediated Akt signal. Cancers 13:1892
Jeon H, Huynh DTN, Baek N, Nguyen TLL, Heo KS (2021) Ginsenoside-Rg2 affects cell growth via regulating ROS-mediated AMPK activation and cell cycle in MCF-7 cells. Phytomedicine 85:153549. doi:https://doi.org/10.1016/j.phymed.2021.153549
Jiang S, Wang Y, Luo L, Shi F, Zou J, Lin H, Ying Y, Luo Y, Zhan Z, Liu P, Zhu B, Huang D, Luo Z (2019) AMP-activated protein kinase regulates cancer cell growth and metabolism via nuclear and mitochondria events. J cell Mol Med 23(6):3951–3961. doi:https://doi.org/10.1111/jcmm.14279
Jin Y, Huynh DTN, Kang KW, Myung CS, Heo KS (2019) Inhibition of p90RSK activation sensitizes triple-negative breast cancer cells to cisplatin by inhibiting proliferation, migration and EMT. BMB Rep 52:706–711
Jin Y, Huynh DTN, Nguyen LLT, Jeon HS, Heo KS (2020) Therapeutic effects of ginsenosides on breast cancer growth and metastasis. Arch Pharm Res 43:773–787. https://doi.org/10.1007/s12272-020-01265-8
Kang SW, Lee S, Lee EK (2015) ROS and energy metabolism in cancer cells: alliance for fast growth. Arch Pharm Res 38:338–345. https://doi.org/10.1007/s12272-015-0550-6
Kim MY, Bo HH, Choi EO, Kwon DH, Kim HJ, Ahn KI, Ji SY, Jeong JW, Park SH, Hong SH, Kim GY, Park C, Kim HS, Moon SK, Yun SJ, Kim WJ, Choi YH (2018) Induction of apoptosis by citrus unshiu peel in human breast cancer MCF-7 cells: involvement of ROS-dependent activation of AMPK. Biol Pharm Bull 41:713–721. https://doi.org/10.1248/bpb.b17-00898
Kong MJ, Han SJ, Kim JI, Park JW, Park KM (2018) Mitochondrial NADP(+)-dependent isocitrate dehydrogenase deficiency increases cisplatin-induced oxidative damage in the kidney tubule cells. Cell Death Dis 9:488. https://doi.org/10.1038/s41419-018-0537-6
Lee YJ, Park KS, Heo SH, Nam HS, Cho MK, Lee SH (2019) Pifithrin-µ induces necroptosis through oxidative mitochondrial damage but accompanies epithelial-mesenchymal transition-like phenomenon in malignant mesothelioma cells under lactic acidosis. Arch Pharm Res 42:890–901. https://doi.org/10.1007/s12272-019-01181-6
Li X, Qu Z, Jing S, Li X, Zhao C, Man S, Wang Y, Gao W (2019) Dioscin-6’-O-acetate inhibits lung cancer cell proliferation via inducing cell cycle arrest and caspase-dependent apoptosis. Phytomedicine 53:124–133. https://doi.org/10.1016/j.phymed.2018.09.033
Liu H, Liu M, Jin Z, Yaqoob S, Zheng M, Cai D, Liu J, Guo S (2019) Ginsenoside Rg2 inhibits adipogenesis in 3T3-L1 preadipocytes and suppresses obesity in high-fat-diet-induced obese mice through the AMPK pathway. Food Funct 10:3603–3614. https://doi.org/10.1039/c9fo00027e
Nguyen TLL, Huynh DTN, Jin Y, Jeon H, Heo KS (2021) Protective effects of ginsenoside-Rg2 and -Rh1 on liver function through inhibiting TAK1 and STAT3-mediated inflammatory activity and Nrf2/ARE-mediated antioxidant signaling pathway. Arch Pharm Res 44:241–252. https://doi.org/10.1007/s12272-020-01304-4
Pena-Blanco A, Garcia-Saez AJ (2018) Bax, Bak and beyond - mitochondrial performance in apoptosis. FEBS J 285:416–431. https://doi.org/10.1111/febs.14186
Pluchino LA, Choudhary S, Wang HC (2016) Reactive oxygen species-mediated synergistic and preferential induction of cell death and reduction of clonogenic resistance in breast cancer cells by combined cisplatin and FK228. Cancer Lett 381:124–132. https://doi.org/10.1016/j.canlet.2016.07.036
Pordeli M, Nakhjiri M, Safavi M, Ardestani SK, Foroumadi A (2017) Anticancer effects of synthetic hexahydrobenzo [g]chromen-4-one derivatives on human breast cancer cell lines. Breast Cancer 24:299–311. https://doi.org/10.1007/s12282-016-0704-5
Shao C, Lu W, Du Y, Yan W, Bao Q, Tian Y, Wang G, Ye H, Hao H (2020) Cytosolic ME1 integrated with mitochondrial IDH2 supports tumor growth and metastasis. Redox Biol 36:101685. https://doi.org/10.1016/j.redox.2020.101685
Shin MK, Cheong JH (2019) Mitochondria-centric bioenergetic characteristics in cancer stem-like cells. Arch Pharm Res 42:113–127. https://doi.org/10.1007/s12272-019-01127-y
Skonieczna M, Hejmo T, Poterala-Hejmo A, Cieslar-Pobuda A, Buldak RJ (2017) NADPH oxidases: insights into selected functions and mechanisms of action in cancer and stem cells. Oxid Med Cell Longev 2017:9420539. https://doi.org/10.1155/2017/9420539
Tan Z, Luo X, Xiao L, Tang M, Bode AM, Dong Z, Cao Y (2016) The role of PGC1alpha in cancer metabolism and its therapeutic implications. Mol Cancer Ther 15:774–782. https://doi.org/10.1158/1535-7163.MCT-15-0621
Wallace DC (2012) Mitochondria and cancer. Nat Rev Cancer 12:685–698. https://doi.org/10.1038/nrc3365
Yi B, Liu D, He M, Li Q, Liu T, Shao J (2013) Role of the ROS/AMPK signaling pathway in tetramethylpyrazine-induced apoptosis in gastric cancer cells. Oncol Lett 6:583–589. https://doi.org/10.3892/ol.2013.1403
Zhang Y, Ba Y, Liu C, Sun G, Ding L, Gao S, Hao J, Yu Z, Zhang J, Zen K, Tong Z, Xiang Y (2007) PGC-1α induces apoptosis in human epithelial ovarian cancer cells through a PPARγ-dependent pathway. Cell Res 17:363–373
Zhang HS, Du GY, Zhang ZG, Zhou Z, Sun HL, Yu XY, Shi YT, Xiong DN, Li H, Huang YH (2018) NRF2 facilitates breast cancer cell growth via HIF1ɑ-mediated metabolic reprogramming. ijBCB 95:85–92. https://doi.org/10.1016/j.biocel
Zhang HS, Zhang ZG, Du GY, Sun HL, Liu HY, Zhou Z, Gou XM, Wu XH, Yu XY, Huang YH (2019) Nrf2 promotes breast cancer cell migration via up-regulation of G6PD/HIF-1α/Notch1 axis. J Cell Mol Med 23(5):3451–3463. https://doi.org/10.1111/jcmm.14241
Zhao Y, Hu X, Liu Y, Dong S, Wen Z, He W, Zhang S, Huang Q, Shi M (2017) ROS signaling under metabolic stress: cross-talk between AMPK and AKT pathway. Mol Cancer 16:79. https://doi.org/10.1186/s12943-017-0648-1
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This research was supported by National Research Foundation of Korea (KNRF-2019R1C1C100733112).
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Jeon, H., Jin, Y., Myung, CS. et al. Ginsenoside-Rg2 exerts anti-cancer effects through ROS-mediated AMPK activation associated mitochondrial damage and oxidation in MCF-7 cells. Arch. Pharm. Res. 44, 702–712 (2021). https://doi.org/10.1007/s12272-021-01345-3
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DOI: https://doi.org/10.1007/s12272-021-01345-3