Open Access

COX‑2 promotes epithelial‑mesenchymal transition and migration in osteosarcoma MG‑63 cells via PI3K/AKT/NF‑κB signaling

  • Authors:
    • Xueliang Zhang
    • Peng Qu
    • Hui Zhao
    • Tong Zhao
    • Nong Cao
  • View Affiliations

  • Published online on: August 21, 2019     https://doi.org/10.3892/mmr.2019.10598
  • Pages: 3811-3819
  • Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to investigate the mechanism by which cyclooxygenase‑2 (COX‑2) promotes the metastasis of MG‑63 osteosarcoma cells through the PI3K/AKT/NF‑κB pathway. To achieve this, a recombinant lentivirus containing the COX‑2 gene was constructed in order to overexpress COX‑2; a recombinant lentivirus containing a control sequence was also constructed. A Transwell chamber migration assay was performed to quantify the migration of the COX‑2‑transduced cells, and of cells treated with a COX‑2 inhibitor (NS398) or a PI3K inhibitor (LY294002). Immunofluorescence assays were performed to determine changes in E‑cadherin, vimentin and NF‑κB expression levels. ELISAs were performed to quantify the levels of matrix metallopeptidase (MMP)‑2, MMP‑9 and vascular endothelial growth factor (VEGF) in the culture medium. Western blot analysis was conducted to measure the protein expression levels of MMP‑2, MMP‑9, PI3K, phosphorylated (p‑) PI3K, AKT, p‑AKT, inhibitor of NF‑κΒ kinase (IKK) and p‑IKK. The results demonstrated that the migration ability of the COX‑2‑overexpressing MG‑63 cells was significantly increased compared with the control cells. The migration ability of cells treated with NS398 or LY294002 was significantly decreased. Compared with the control cells, E‑cadherin expression was significantly decreased in COX‑2‑overexpressing cells, while the expression levels of vimentin, MMP‑2, MMP‑9, VEGF, p‑PI3K, p‑AKT and p‑IKK were significantly increased. Compared with the control cells, E‑cadherin expression was significantly increased in cells treated with NS398 or LY294002, while the expression levels of vimentin, MMP‑2, MMP‑9, VEGF, p‑PI3K, p‑AKT, and p‑IKK were significantly decreased. The total protein levels of PI3K, AKT and IKK were not changed among the treatment groups. In summary, COX‑2 overexpression decreased the expression levels of the epithelial protein E‑cadherin and increased the expression levels of the mesenchymal proteins vimentin, MMP‑2 and MMP‑9, as well as promoted cell migration, by activating the PI3K/AKT/NF‑κB signaling pathway.

References

1 

He X, Gao Z, Xu H, Zhang Z and Fu P: A meta-analysis of randomized control trials of surgical methods with osteosarcoma outcomes. J Orthop Surg Res. 12:52017. View Article : Google Scholar : PubMed/NCBI

2 

Isakoff MS, Bielack SS, Meltzer P and Gorlick R: Osteosarcoma: Current treatment and a collaborative pathway to success. J Clin Oncol. 33:3029–3035. 2015. View Article : Google Scholar : PubMed/NCBI

3 

Geller DS and Gorlick R: Osteosarcoma: A review of diagnosis, management, and treatment strategies. Clin Adv Hematol Oncol. 8:705–718. 2010.PubMed/NCBI

4 

Kalluri R and Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest. 119:1420–1428. 2009. View Article : Google Scholar : PubMed/NCBI

5 

Morel AP, Hinkal GW, Thomas C, Fauvet F, Courtois-Cox S, Wierinckx A, Devouassoux-Shisheboran M, Treilleux I, Tissier A, Gras B, et al: EMT inducers catalyze malignant transformation of mammary epithelial cells and drive tumorigenesis towards claudin-low tumors in transgenic mice. PLoS Genet. 8:e10027232012. View Article : Google Scholar : PubMed/NCBI

6 

Amatangelo MD, Goodyear S and Varma D: c-Myc expression and MEK1-induced Erk2 nuclear localization are required for TGF-beta induced epithelial-mesenchymal transition and invasion in prostate cancer. Carcinogenesis. 33:1965–1975. 2012. View Article : Google Scholar : PubMed/NCBI

7 

Sharili AS, Allen S, Smith K, Price J and McGonnell IM: Snail2 promotes osteosarcoma cell motility through remodelling of the actin cytoskeleton and regulates tumor development. Cancer Lett. 333:170–179. 2013. View Article : Google Scholar : PubMed/NCBI

8 

Tsujii M, Kawano S and Dubois RN: Cyclooxygenase-2 expression in human colon cancer cells increases metastatic potential. Proc Natl Acad Sci USA. 94:3336–3340. 1997. View Article : Google Scholar : PubMed/NCBI

9 

Tsujii M, Kawano S, Tsuji S, Sawaoka H, Hori M and DuBois RN: Cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell. 93:705–716. 1998. View Article : Google Scholar : PubMed/NCBI

10 

Masi L, Recenti R, Silvestri S, Pinzani P, Pepi M, Paglierani M, Brandi ML and Franchi A: Expression of cyclooxygenase-2 in osteosarcoma of bone. Appl Immunohistochem Mol Morphol. 15:70–76. 2007. View Article : Google Scholar : PubMed/NCBI

11 

Rodriguez NI, Hoots WK, Koshkina NV, Morales-Arias JA, Arndt CA, Inwards CY, Hawkins DS, Munsell MF and Kleinerman ES: COX-2 expression correlates with survival in patients with osteosarcoma lung metastases. J Pediatr Hematol Oncol. 30:507–512. 2008. View Article : Google Scholar : PubMed/NCBI

12 

Pang LY, Gatenby EL, Kamida A, Whitelaw BA, Hupp TR and Argyle DJ: Global gene expression analysis of canine osteosarcoma stem cells reveals a novel role for COX-2 in tumour initiation. PLoS One. 9:e831442014. View Article : Google Scholar : PubMed/NCBI

13 

Lee EJ, Choi EM, Kim SR, Park JH, Kim H, Ha KS, Kim YM, Kim SS, Choe M, Kim JI and Han JA: Cyclooxygenase-2 promotes cell proliferation, migration and invasion in U2OS human osteosarcoma cells. Exp Mol Med. 39:469–476. 2007. View Article : Google Scholar : PubMed/NCBI

14 

Urakawa H, Nishida Y, Naruse T, Nakashima H and Ishiguro N: Cyclooxygenase-2 overexpression predicts poor survival in patients with high-grade extremity osteosarcoma: A pilot study. Clin Orthop Relat Res. 467:2932–2938. 2009. View Article : Google Scholar : PubMed/NCBI

15 

Ji C, Guo H, Zhang P, Kuang W, Fan Y and Wu L: AnnexinA5 promote glioma cell invasion and migration via the PI3K/Akt/NF-κB signaling pathway. J Neurooncol. 138:469–478. 2018. View Article : Google Scholar : PubMed/NCBI

16 

Zhu LB, Jiang J, Zhu XP, Wang TF, Chen XY, Luo QF, Shu Y, Liu ZL and Huang SH: Knockdown of Aurora-B inhibits osteosarcoma cell invasion and migration via modulating PI3K/Akt/NF-κB signaling pathway. Int J Clin Exp Pathol. 7:3984–3991. 2014.PubMed/NCBI

17 

Dong Y, Liang G, Yuan B, Yang C, Gao R and Zhou X: MALAT1 promotes the proliferation and metastasis of osteosarcoma cells by activating the PI3K/Akt pathway. Tumour Biol. 36:1477–1486. 2015. View Article : Google Scholar : PubMed/NCBI

18 

He ML, Wu Y, Zhao JM, Wang Z and Chen YB: PIK3CA and AKT gene polymorphisms in susceptibility to osteosarcoma in a Chinese population. Asian Pac J Cancer Prev. 14:5117–5122. 2013. View Article : Google Scholar : PubMed/NCBI

19 

Orgaz JL, Ladhani O, Hoek KS, Fernández-Barral A, Mihic D, Aguilera O, Seftor EA, Bernad A, Rodríguez-Peralto JL, Hendrix MJ, et al: ‘Loss of pigment epithelium-derived factor enables migration, invasion and metastatic spread of human melanoma’. Oncogene. 28:4147–4161. 2009. View Article : Google Scholar : PubMed/NCBI

20 

Luan W, Yao Q, Xin N, Bu X, Xia Y, Wang J, Ruan H, Ma S and Xu B: miR-204-5p acts as a tumor suppressor by targeting matrix metalloproteinases-9 and B-cell lymphoma-2 in malignant melanoma. Onco Targets Ther. 10:1237–1246. 2017. View Article : Google Scholar : PubMed/NCBI

21 

Vincent CT and Fuxe J: EMT, inflammation and metastasis. Semin Cancer Biol. 47:168–169. 2017. View Article : Google Scholar : PubMed/NCBI

22 

Wu Y and Zhou BP: New insights of epithelial-mesenchymal transition in cancer metastasis. Acta Biochim Biophys Sin (Shanghai). 40:643–650. 2008. View Article : Google Scholar : PubMed/NCBI

23 

Amano S, Akutsu N, Matsunaga Y, Nishiyama T, Champliaud MF, Burgeson RE and Adachi E: Importance of balance between extracellular matrix synthesis and degradation in basement membrane formation. Exp Cell Res. 271:249–262. 2001. View Article : Google Scholar : PubMed/NCBI

24 

Liao CL, Chu YL, Lin HY, Chen CY, Hsu MJ, Liu KC, Lai KC, Huang AC and Chung JG: Bisdemethoxycurcumin suppresses migration and invasion of human cervical cancer HeLa cells via inhibition of NF-ĸB, MMP-2 and −9 pathways. Anticancer Res. 38:3989–3997. 2018. View Article : Google Scholar : PubMed/NCBI

25 

Wu S, Yang D, Beckford J and Alachkar H: Upregulation of the EMT marker vimentin is associated with poor clinical outcome in acute myeloid leukemia. J Transl Med. 16:1702018. View Article : Google Scholar : PubMed/NCBI

26 

Gou Y, Zhai F, Zhang L and Cui L: RUNX3 regulates hepatocellular carcinoma cell metastasis via targeting miR-186/E-cadherin/EMT pathway. Oncotarget. 8:61475–61486. 2017. View Article : Google Scholar : PubMed/NCBI

27 

Zhao G, Cai C, Yang T, Qiu X, Liao B, Li W, Ji Z, Zhao J, Zhao H, Guo M, et al: MicroRNA-221 induces cell survival and cisplatin resistance through PI3K/Akt pathway in human osteosarcoma. PLoS One. 8:e539062013. View Article : Google Scholar : PubMed/NCBI

28 

Guo YS, Zhao R, Ma J, Cui W, Sun Z, Gao B, He S, Han YH, Fan J, Yang L, et al: βig-h3 promotes human osteosarcoma cells metastasis by interacting with integrin α2β1 and activating PI3K signaling pathway. PLoS One. 9:e902202014. View Article : Google Scholar : PubMed/NCBI

29 

Hou CH, Lin FL, Tong KB, Hou SM and Liu JF: Transforming growth factor alpha promotes osteosarcoma metastasis by ICAM-1 and PI3K/Akt signaling pathway. Biochem Pharmacol. 89:453–463. 2014. View Article : Google Scholar : PubMed/NCBI

30 

Aizawa J, Sakayama K, Kamei S, Kidani T, Yamamoto H, Norimatsu Y and Masuno H: Effect of troglitazone on tumor growth and pulmonary metastasis development of the mouse osteosarcoma cell line LM8. BMC Cancer. 10:512010. View Article : Google Scholar : PubMed/NCBI

31 

Tsubaki M, Satou T, Itoh T, Imano M, Ogaki M, Yanae M and Nishida S: Reduction of metastasis, cell invasion, and adhesion in mouse osteosarcoma by YM529/ONO-5920-induced blockade of the Ras/MEK/ERK and Ras/PI3K/Akt pathway. Toxicol Appl Pharmacol. 259:402–410. 2012. View Article : Google Scholar : PubMed/NCBI

32 

Hayden MS and Ghosh S: Signaling to NF-kappaB. Genes Dev. 18:2195–2224. 2004. View Article : Google Scholar : PubMed/NCBI

33 

Li J, Lau GK, Chen L, Dong SS, Lan HY, Huang XR, Li Y, Luk JM, Yuan YF and Guan XY: Interleukin 17A promotes hepatocellular carcinoma metastasis via NF-kB induced matrix metalloproteinases 2 and 9 expression. PLoS One. 6:e218162011. View Article : Google Scholar : PubMed/NCBI

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October 2019
Volume 20 Issue 4

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APA
Zhang, X., Qu, P., Zhao, H., Zhao, T., & Cao, N. (2019). COX‑2 promotes epithelial‑mesenchymal transition and migration in osteosarcoma MG‑63 cells via PI3K/AKT/NF‑κB signaling. Molecular Medicine Reports, 20, 3811-3819. https://doi.org/10.3892/mmr.2019.10598
MLA
Zhang, X., Qu, P., Zhao, H., Zhao, T., Cao, N."COX‑2 promotes epithelial‑mesenchymal transition and migration in osteosarcoma MG‑63 cells via PI3K/AKT/NF‑κB signaling". Molecular Medicine Reports 20.4 (2019): 3811-3819.
Chicago
Zhang, X., Qu, P., Zhao, H., Zhao, T., Cao, N."COX‑2 promotes epithelial‑mesenchymal transition and migration in osteosarcoma MG‑63 cells via PI3K/AKT/NF‑κB signaling". Molecular Medicine Reports 20, no. 4 (2019): 3811-3819. https://doi.org/10.3892/mmr.2019.10598