Oleanolic acid inhibits osteosarcoma cell proliferation and invasion by suppressing the SOX9/Wnt1 signaling pathway

Chen,Xianming; Zhang,Yulin; Zhang,Sen; Wang,Aiming; Du,Quanyin; Wang,Ziming

doi:10.3892/etm.2021.9883

Oleanolic acid inhibits osteosarcoma cell proliferation and invasion by suppressing the SOX9/Wnt1 signaling pathway

Authors:
- Xianming Chen
- Yulin Zhang
- Sen Zhang
- Aiming Wang
- Quanyin Du
- Ziming Wang
View Affiliations

Affiliations: Department of Osteopathic Medicine, Daping Hospital, Army Medical University, Chongqing 400042, P.R. China, Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital and Chongqing Cancer Institute and Chongqing Cancer Hospital, Chongqing 400030, P.R. China
Published online on: February 28, 2021 https://doi.org/10.3892/etm.2021.9883
Article Number: 443
Copyright: © Chen 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

Osteosarcoma is the most common primary bone malignancy in children and adolescents. Inhibition of SOX9/Wnt1‑mediated signaling might suppress osteosarcoma metastasis, and oleanolic acid (OA) might decrease the activity of the SOX9/Wnt1 signaling pathway. The aim of the present study was to determine the role of OA in osteosarcoma cell proliferation and invasion. Osteosarcoma cell lines (KHOS and U2OS) and an osteoblastic cell line (hFOB1.19) were used for cell viability, proliferation and invasion analysis. The data suggested that OA significantly inhibited cell viability on days 3, 4 and 5 compared with the control (Ctrl) group in both U2OS and KHOS cells. Cell proliferation in the OA‑treated group was significantly decreased compared with the Ctrl group in the osteosarcoma cell lines. Analysis of the cell cycle indicated that OA significantly reduced the percentage of U2OS and KHOS cells in the S phase compared with the Ctrl group. The wound healing assay results indicated that the OA group displayed significantly decreased cell re‑colonization of the wound at 48 h compared with the Ctrl group. The Transwell chamber assay results also indicated that cell invasion was significantly inhibited by OA compared with the Ctrl group. Furthermore, OA significantly increased osteosarcoma cell apoptosis compared with the Ctrl group. Similarly, the protein expression levels of SOX9 and Wnt1 were significantly decreased in OA‑treated U2OS and KHOS cells compared with Ctrl cells. OA‑mediated downregulation of Wnt1 expression was reversed following SOX9 small interfering RNA transfection. Collectively, the results indicated that OA inhibited SOX9/Wnt1‑associated osteosarcoma cell proliferation, migration and invasion.

View Figures

View References

1	Sadykova LR, Ntekim AI, Muyangwa-Semenova M, Rutland CS, Jeyapalan JN, Blatt N and Rizvanov AA: Epidemiology and risk factors of osteosarcoma. Cancer Invest. 38:259–269. 2020.PubMed/NCBI View Article : Google Scholar
2	Morrow JJ, Bayles I, Funnell APW, Miller TE, Saiakhova A, Lizardo MM, Bartels CF, Kapteijn MY, Hung S, Mendoza A, et al: Positively selected enhancer elements endow osteosarcoma cells with metastatic competence. Nat Med. 24:176–185. 2018.PubMed/NCBI View Article : Google Scholar
3	Liu H, Chen Y, Zhou F, Jie L, Pu L, Ju J, Li F, Dai Z, Wang X and Zhou S: Sox9 regulates hyperexpression of Wnt1 and Fzd1 in human osteosarcoma tissues and cells. Int J Clin Exp Pathol. 7:4795–4805. 2014.PubMed/NCBI
4	Wang S, Zhang D, Han S, Gao P, Liu C, Li J and Pan X: Fibulin-3 promotes osteosarcoma invasion and metastasis by inducing epithelial to mesenchymal transition and activating the Wnt/β-catenin signaling pathway. Sci Rep. 7(6215)2017.PubMed/NCBI View Article : Google Scholar
5	Zou J, Zhang W and Li XL: Effects of SOST gene silencing on proliferation, apoptosis, invasion, and migration of human osteosarcoma cells through the wnt/β-catenin signaling pathway. Calcif Tissue Int. 100:551–564. 2017.PubMed/NCBI View Article : Google Scholar
6	Xu Y, Shu B, Tian Y, Wang G, Wang Y, Wang J and Dong Y: Oleanolic acid induces osteosarcoma cell apoptosis by inhibition of Notch signaling. Mol Carcinog. 57:896–902. 2018.PubMed/NCBI View Article : Google Scholar
7	Gao F, Zuo Q, Jiang T, Song H and Zhou J: A newly synthesized oleanolic acid derivative inhibits the growth of osteosarcoma cells in vitro and in vivo by decreasing c-MYC-dependent glycolysis. J Cell Biochem. 120:9264–9276. 2019.PubMed/NCBI View Article : Google Scholar
8	Shanmugam MK, Dai X, Kumar AP, Tan BK, Sethi G and Bishayee A: Oleanolic acid and its synthetic derivatives for the prevention and therapy of cancer: Preclinical and clinical evidence. Cancer Lett. 346:206–216. 2014.PubMed/NCBI View Article : Google Scholar
9	Fan X, Wang P, Sun Y, Jiang J, Du H, Wang Z, Duan Z, Lei H and Li H: Oleanolic acid derivatives inhibit the Wnt/β-catenin signaling pathway by promoting the phosphorylation of beta-catenin in human SMMC-7721 cells. Pharmazie. 71:398–401. 2016.PubMed/NCBI View Article : Google Scholar
10	Wang YY, Yang YX, Zhao R, Pan ST, Zhe H, He ZX, Duan W, Zhang X, Yang T, Qiu JX and Zhou SF: Bardoxolone methyl induces apoptosis and autophagy and inhibits epithelial-to-mesenchymal transition and stemness in esophageal squamous cancer cells. Drug Des Devel Ther. 9:993–1026. 2015.PubMed/NCBI View Article : Google Scholar
11	Landers JE, Cassel SL and George DL: Translational enhancement of mdm2 oncogene expression in human tumor cells containing a stabilized wild-type p53 protein. Cancer Res. 57:3562–3568. 1997.PubMed/NCBI
12	Rhim JS, Cho HY, Vernon ML, Arnstein P, Huebner RJ and Gilden RV: Characterization of non-producer human cells induced by Kirsten sarcoma virus. Int J Cancer. 16:840–849. 1975.PubMed/NCBI View Article : Google Scholar
13	Duan L, Yang Z, Jiang X, Zhang J and Guo X: Oleanolic acid inhibits cell proliferation migration and invasion and induces SW579 thyroid cancer cell line apoptosis by targeting forkhead transcription factor A. Anticancer Drugs. 30:812–820. 2019.PubMed/NCBI View Article : Google Scholar
14	Hotchkiss RS, Strasser A, McDunn JE and Swanson PE: Cell death. N Engl J Med. 361:1570–1583. 2009.PubMed/NCBI View Article : Google Scholar
15	Zhang SL, Yang ZN, He C, Liao HB, Wang HS, Chen ZF and Liang D: Oleanane-type triterpenoid saponins from lysimachia fortunei maxim. Phytochemistry. 147:140–146. 2018.PubMed/NCBI View Article : Google Scholar
16	Peng XP, Li XH, Li Y, Huang XT and Luo ZQ: The protective effect of oleanolic acid on NMDA-induced MLE-12 cells apoptosis and lung injury in mice by activating SIRT1 and reducing NF-kB acetylation. Int Immunopharmacol. 70:520–529. 2019.PubMed/NCBI View Article : Google Scholar
17	Žiberna L, Šamec D, Mocan A, Nabavi SF, Bishayee A, Farooqi AA, Sureda A and Nabavi SM: Oleanolic acid alters multiple cell signaling pathways: Implication in cancer prevention and therapy. Int J Mol Sci. 18(643)2017.PubMed/NCBI View Article : Google Scholar
18	Zhu YY, Huang HY and Wu YL: Anticancer and apoptotic activities of oleanolic acid are mediated through cell cycle arrest and disruption of mitochondrial membrane potential in HepG2 human hepatocellular carcinoma cells. Mol Med Rep. 12:5012–5018. 2015.PubMed/NCBI View Article : Google Scholar
19	Liese J, Hinrichs TM, Lange M and Fulda S: Cotreatment with sorafenib and oleanolic acid induces reactive oxygen species-dependent and mitochondrial-mediated apoptotic cell death in hepatocellular carcinoma cells. Anticancer Drugs. 30:209–217. 2019.PubMed/NCBI View Article : Google Scholar
20	Kim GJ, Jo HJ, Lee KJ, Choi JW and An JH: Oleanolic acid induces p53-dependent apoptosis via the ERK/JNK/AKT pathway in cancer cell lines in prostatic cancer xenografts in mice. Oncotarget. 9:26370–26386. 2018.PubMed/NCBI View Article : Google Scholar
21	Zhu B, Ren C, Du K, Zhu H, Ai Y, Kang F, Luo Y, Liu W, Wang L, Xu Y, et al: Olean-28,13b-olide 2 plays a role in cisplatin-mediated apoptosis and reverses cisplatin resistance in human lung cancer through multiple signaling pathways. Biochem Pharmacol. 170(113642)2019.PubMed/NCBI View Article : Google Scholar
22	Khan MW, Zhao P, Khan A, Raza F, Raza SM, Sarfraz M, Chen Y, Li M, Yang T, Ma X and Xiang G: Synergism of cisplatin-oleanolic acid co-loaded calcium carbonate nanoparticles on hepatocellular carcinoma cells for enhanced apoptosis and reduced hepatotoxicity. Int J Nanomedicine. 14:3753–3771. 2019.PubMed/NCBI View Article : Google Scholar
23	Shi Y, Song Q, Hu D, Zhuang X, Yu S and Teng D: Oleanolic acid induced autophagic cell death in hepatocellular carcinoma cells via PI3K/Akt/mTOR and ROS-dependent pathway. Korean J Physiol Pharmacol. 20:237–243. 2016.PubMed/NCBI View Article : Google Scholar
24	Nie H, Wang Y, Qin Y and Gong XG: Oleanolic acid induces autophagic death in human gastric cancer cells in vitro and in vivo. Cell Biol Int. 40:770–778. 2016.PubMed/NCBI View Article : Google Scholar
25	Khurana N and Sikka SC: Interplay between SOX9, Wnt/β-catenin and androgen receptor signaling in castration-resistant prostate cancer. Int J Mol Sci. 20(2066)2019.PubMed/NCBI View Article : Google Scholar
26	Yu Y, Yin W, Yu ZH, Zhou YJ, Chi JR, Ge J and Cao XC: miR-190 enhances endocrine therapy sensitivity by regulating SOX9 expression in breast cancer. J Exp Clin Cancer Res. 38(22)2019.PubMed/NCBI View Article : Google Scholar
27	Domenici G, Aurrekoetxea-Rodriguez I, Simoes BM, Rábano M, Lee SY, Millán JS, Comaills V, Oliemuller E, López-Ruiz JA, Zabalza I, et al: A Sox2-Sox9 signalling axis maintains human breast luminal progenitor and breast cancer stem cells. Oncogene. 38:3151–3169. 2019.PubMed/NCBI View Article : Google Scholar
28	Suh N, Paul S, Lee HJ, Yoon T, Shah N, Son AI, Reddi AH, Medici D and Sporn MB: Synthetic triterpenoids, CDDO-Imidazolide and CDDO-Ethyl amide, induce chondrogenesis. Osteoarthritis Cartilage. 20:446–450. 2012.PubMed/NCBI View Article : Google Scholar
29	Borella R, Forti L, Gibellini L, De Gaetano A, De Biasi S, Nasi M, Cossarizza A and Pinti M: Synthesis and anticancer activity of CDDO and CDDO-Me, two derivatives of natural triterpenoids. Molecules. 24(4097)2019.PubMed/NCBI View Article : Google Scholar
30	Lin C, Wen X and Sun H: Oleanolic acid derivatives for pharmaceutical use: A patent review. Expert Opin Ther Pat. 26:643–655. 2016.PubMed/NCBI View Article : Google Scholar
31	Oh HJ, Kido T and Lau YF: PIAS1 interacts with and represses SOX9 transactivation activity. Mol Reprod Dev. 74:1446–1455. 2007.PubMed/NCBI View Article : Google Scholar
32	Hattori T, Eberspaecher H, Lu J, Zhang R, Nishida T, Kahyo T, Yasuda H and de Crombrugghe B: Interactions between PIAS proteins and SOX9 result in an increase in the cellular concentrations of SOX9. J Biol Chem. 281:14417–14428. 2006.PubMed/NCBI View Article : Google Scholar
33	Saotome H, Ito A, Kubo A and Inui M: Generation of a quantitative luciferase reporter for Sox9 SUMOylation. Int J Mol Sci. 21(1274)2020.PubMed/NCBI View Article : Google Scholar
34	Wu J, Lei H, Zhang J, Chen X, Tang C, Wang W, Xu H, Xiao W, Gu W and Wu Y: Momordin Ic, a new natural SENP1 inhibitor, inhibits prostate cancer cell proliferation. Oncotarget. 7:58995–59005. 2016.PubMed/NCBI View Article : Google Scholar