Calycosin induces apoptosis via p38‑MAPK pathway‑mediated activation of the mitochondrial apoptotic pathway in human osteosarcoma 143B cells

Tian,Wei; Wang,Zhi‑Wei; Yuan,Bao‑Ming; Bao,Yong‑Ge

doi:10.3892/mmr.2020.11471

Calycosin induces apoptosis via p38‑MAPK pathway‑mediated activation of the mitochondrial apoptotic pathway in human osteosarcoma 143B cells

Authors:
- Wei Tian
- Zhi‑Wei Wang
- Bao‑Ming Yuan
- Yong‑Ge Bao
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Affiliations: Department of Orthopedics, Affiliated Hospital of Inner Mongolia University for The Nationalities, Tongliao, Inner Mongolia 028007, P.R. China
Published online on: September 1, 2020 https://doi.org/10.3892/mmr.2020.11471
Pages: 3962-3968
Copyright: © Tian et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Previous studies have demonstrated that calycosin is a natural phytoestrogen with a similar structure to estrogen, which can inhibit cell proliferation and induce apoptosis in a variety of tumors. Calycosin exerts potential pharmacological effects on osteosarcoma cells by inducing apoptosis. The aim of the present study was to elucidate the specific molecular mechanism of calycosin‑induced apoptosis in osteosarcoma cells. Cell proliferation was determined by an MTT assay. Annexin V/PI and JC‑1 staining were used to detect apoptosis and mitochondrial dysfunction, respectively, by flow cytometry. Western blot analysis was used to detect the expression of caspases or mitochondrial proteins. The results revealed that calycosin reduced the cell viability of human osteosarcoma 143B cells, induced apoptosis and increased the loss of mitochondrial membrane potential (MMP). In addition, calycosin increased the expression of the proapoptotic antiapoptotic proteins cleaved caspase‑3, cleaved caspase‑9, cleaved poly(ADP‑ribose) polymerase and Bcl‑2‑associated X protein (Bax), and decreased the expression of the antiapoptotic proapoptotic protein B‑cell lymphoma‑2 (Bcl‑2), thus altering the Bax/Bcl‑2 ratio. In addition, the expression levels of cytochrome c were markedly decreased in the mitochondria and increased in the cytoplasm following calycosin treatment. Furthermore, calycosin treatment induced p38‑mitogen‑activated protein kinase (MAPK) phosphorylation, whereas the p38‑MAPK inhibitor BIRB 796 markedly reversed cell viability, apoptosis and loss of MMP in 143B cells. These results suggested that calycosin inhibited osteosarcoma 143B cell growth via p38‑MAPK regulation of mitochondrial‑dependent intrinsic apoptotic pathways.

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1	Mirabello L, Troisi RJ and Savage SA: Osteosarcoma incidence and survival rates from 1973 to 2004: Data from the surveillance, epidemiology, and end results program. Cancer. 115:1531–1543. 2009. View Article : Google Scholar : PubMed/NCBI
2	Eyre R, Feltbower RG, Mubwandarikwa E, Eden TO and McNally RJ: Epidemiology of bone tumours in children and young adults. Pediatr Blood Cancer. 53:941–952. 2009. View Article : Google Scholar : PubMed/NCBI
3	Harrison DJ, Geller DS, Gill JD, Lewis VO and Gorlick R: Current and future therapeutic approaches for osteosarcoma. Expert Rev Anticancer Ther. 18:39–50. 2018. View Article : Google Scholar : PubMed/NCBI
4	Baudino TA: Targeted cancer therapy: The next generation of cancer treatment. Curr Drug Discov Technol. 12:3–20. 2015. View Article : Google Scholar : PubMed/NCBI
5	Tao Y, Wang Y, Wang X, Wang C, Bao K, Ji L, Jiang G and Hong M: Calycosin suppresses epithelial derived initiative key factors and maintains epithelial barrier in allergic inflammation via TLR4 mediated NF-κB pathway. Cell Physiol Biochem. 44:1106–1119. 2017. View Article : Google Scholar : PubMed/NCBI
6	Dong L, Yin L, Chen R, Zhang Y, Hua S, Quan H and Fu X: Anti-inflammatory effect of Calycosin glycoside on lipopolysaccharide-induced inflammatory responses in RAW 264.7 cells. Gene. 675:94–101. 2018. View Article : Google Scholar : PubMed/NCBI
7	Duan X, Meng Q, Wang C, Liu Z, Liu Q, Sun H, Sun P, Yang X, Huo X, Peng J and Liu K: Calycosin attenuates triglyceride accumulation and hepatic fibrosis in murine model of non-alcoholic steatohepatitis via activating farnesoid X receptor. Phytomedicine. 25:83–92. 2017. View Article : Google Scholar : PubMed/NCBI
8	Liu Y, Che G, Di Z, Sun W, Tian J and Ren M: Calycosin-7-O-β-D-glucoside attenuates myocardial ischemia-reperfusion injury by activating JAK2/STAT3 signaling pathway via the regulation of IL-10 secretion in mice. Mol Cell Biochem. 463:175–187. 2020. View Article : Google Scholar : PubMed/NCBI
9	Qiu R, Li X, Qin K, Chen X, Wang R, Dai Y, Deng L and Ye Y: Antimetastatic effects of calycosin on osteosarcoma and the underlying mechanism. Biofactors. 45:975–982. 2019. View Article : Google Scholar : PubMed/NCBI
10	Wu G, Niu M, Qin J, Wang Y and Tian J: Inactivation of Rab27B-dependent signaling pathway by calycosin inhibits migration and invasion of ER-negative breast cancer cells. Gene. 709:48–55. 2019. View Article : Google Scholar : PubMed/NCBI
11	El-Kott AF, Al-Kahtani MA and Shati AA: Calycosin induces apoptosis in adenocarcinoma HT29 cells by inducing cytotoxic autophagy mediated by SIRT1/AMPK-induced inhibition of Akt/mTOR. Clin Exp Pharmacol Physiol. 46:944–954. 2019. View Article : Google Scholar : PubMed/NCBI
12	Sun H, Yin M, Qian W and Yin H: Calycosin, a phytoestrogen isoflavone, induces apoptosis of estrogen receptor-positive MG-63 osteosarcoma cells via the phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway. Med Sci Monit. 24:6178–6186. 2018. View Article : Google Scholar : PubMed/NCBI
13	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
14	Ning L, Wan S, Jie Z, Xie Z, Li X, Pan X, Wan X, Chen W, Huang H, Wang J, et al: Lycorine induces apoptosis and G1 phase arrest through ROS/p38 MAPK signaling pathway in human osteosarcoma cells in vitro and in vivo. Spine (Phila Pa 1976). 45:E126–E139. 2019. View Article : Google Scholar
15	Chen J, Hou R, Zhang X, Ye Y, Wang Y and Tian J: Calycosin suppresses breast cancer cell growth via ERβ-dependent regulation of IGF-1R, p38 MAPK and PI3K/Akt pathways. PLoS One. 9:e912452014. View Article : Google Scholar : PubMed/NCBI
16	Zhang B, Wang D, Guo F and Xuan C: Mitochondrial membrane potential and reactive oxygen species in cancer stem cells. Fam Cancer. 14:19–23. 2015. View Article : Google Scholar : PubMed/NCBI
17	Wada T and Penninger JM: Mitogen-activated protein kinases in apoptosis regulation. Oncogene. 23:2838–2849. 2004. View Article : Google Scholar : PubMed/NCBI
18	Qiu R, Ma G, Li X, Shi Q, Li X, Zhou X, Tang Y, Xie Z, Liao S, Qin Y, et al: Clinical case report of patients with osteosarcoma and anticancer benefit of calycosin against human osteosarcoma cells. J Cell Biochem. 120:10697–10706. 2019. View Article : Google Scholar : PubMed/NCBI
19	Fleischer A, Ghadiri A, Dessauge F, Duhamel M, Rebollo MP, Alvarez-Franco F and Rebollo A: Modulating apoptosis as a target for effective therapy. Mol Immunol. 43:1065–1079. 2006. View Article : Google Scholar : PubMed/NCBI
20	Creagh EM: Caspase crosstalk: Integration of apoptotic and innate immune signalling pathways. Trends Immunol. 35:631–640. 2014. View Article : Google Scholar : PubMed/NCBI
21	Zhou Y, Liu QH, Liu CL and Lin L: Calycosin induces apoptosis in human ovarian cancer SKOV3 cells by activating caspases and Bcl-2 family proteins. Tumour Biol. 36:5333–1339. 2015. View Article : Google Scholar : PubMed/NCBI
22	Palmer CS, Osellame LD, Stojanovski D and Ryan MT: The regulation of mitochondrial morphology: Intricate mechanisms and dynamic machinery. Cell Signal. 23:1534–1545. 2011. View Article : Google Scholar : PubMed/NCBI
23	Xiong S, Mu T, Wang G and Jiang X: Mitochondria-mediated apoptosis in mammals. Protein Cell. 5:737–749. 2014. View Article : Google Scholar : PubMed/NCBI
24	Siu WP, Pun PB, Latchoumycandane C and Boelsterli UA: Bax-mediated mitochondrial outer membrane permeabilization (MOMP), distinct from the mitochondrial permeability transition, is a key mechanism in diclofenac-induced hepatocyte injury: Multiple protective roles of cyclosporin A. Toxicol Appl Pharmacol. 227:451–461. 2008. View Article : Google Scholar : PubMed/NCBI
25	Jergens A, Young J, Moore D, Wang C, Hostetter J, Augustine L, Allenspach K, Schmitz S and Mosher C: Bcl-2/Caspase 3 mucosal imbalance favors T cell resistance to apoptosis in dogs with inflammatory bowel disease. Vet Immunol Immunopathol. 158:167–74. 2014. View Article : Google Scholar : PubMed/NCBI
26	Xu T, Wang NS, Fu LL, Ye CY, Yu SQ and Mei CL: Celecoxib inhibits growth of human autosomal dominant polycystic kidney cyst-lining epithelial cells through the VEGF/Raf/MAPK/ERK signaling pathway. Mol Biol Rep. 39:7743–7753. 2012. View Article : Google Scholar : PubMed/NCBI
27	Cuenda A and Rousseau S: p38 MAP-kinases pathway regulation, function and role in human diseases. Biochim Biophys Acta. 1773:1358–1375. 2007. View Article : Google Scholar : PubMed/NCBI
28	Lin RC, Yang SF, Chiou HL, Hsieh SC, Wen SH, Lu KH and Hsieh YH: Licochalcone a-induced apoptosis through the activation of p38MAPK pathway mediated mitochondrial pathways of apoptosis in human osteosarcoma cells in vitro and in vivo. Cells. 8:14412019. View Article : Google Scholar
29	Li Z, Tang X, Luo Y, Chen B, Zhou C, Wu X, Tang Z, Qi X, Cao G, Hao J, et al: NK007 helps in mitigating paclitaxel resistance through p38MAPK activation and HK2 degradation in ovarian cancer. J Cell Physiol. Feb 20–2019.(Epub ahead of print). doi: 10.1002/jcp.28278.
30	Zhang J, Li J, Song H, Xiong Y, Liu D and Bai X: Hydroxysafflor yellow A suppresses angiogenesis of hepatocellular carcinoma through inhibition of p38 MAPK phosphorylation. Biomed Pharmacother. 109:806–814. 2019. View Article : Google Scholar : PubMed/NCBI