Open Access

Expression and significance of Annexin A3 in the osteosarcoma cell lines HOS and U2OS

  • Authors:
    • Xinxin Zeng
    • Shengtao Wang
    • Peng Gui
    • Hao Wu
    • Zhaoxu Li
  • View Affiliations

  • Published online on: July 23, 2019     https://doi.org/10.3892/mmr.2019.10513
  • Pages: 2583-2590
  • Copyright: © Zeng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Annexin A3 (ANXA3) is highly expressed in different types of cancers, but the impact of ANXA3 in bone tumors is still not clear. In the present study, the expression of ANXA3 in osteosarcoma cells was first confirmed by cellular immunofluorescence. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis were used to detect the expression of ANXA3 in osteoblasts in the osteosarcoma cell lines U2OS and HOS. Furthermore, small interfering (si)‑RNA were transfected into U2OS and HOS cells via a liposome‑mediated method. Then once ANXA3 had been successfully downregulated in U2OS and HOS cells, the cells were collected and total protein was extracted after 48 h of transfection. Western blot analysis was used to confirm successful ANXA3 transfection into osteosarcoma cells and the apoptotic rate of HOS and U2OS was detected by flow cytometry. The expression of ANXA3 in the osteosarcoma cell lines HOS and U2OS were first observed by confocal laser scanning microscopy, and was then detected by RT‑qPCR and western blotting. The mRNA and protein levels of ANXA3 in the osteosarcoma cell lines HOS and U2OS were significantly increased compared with osteoblasts, particularly in HOS cells. When siRNA was transfected into HOS and U2OS cells, the protein expression level of ANXA3 was measured via western blotting. The results indicated that the expression of ANXA3 was significantly decreased. In addition, to determine whether ANXA3 knockdown induced cell apoptosis, the present study analyzed the apoptotic rate by flow cytometry. The results revealed that ANXA3 knockdown markedly increased HOS and U2OS cell apoptosis. To the best of our knowledge, the present study is the first to confirm that ANXA3 is highly expressed in the osteosarcoma cell lines HOS and U2OS. In addition, downregulation of ANXA3 expression in HOS and U2OS cells could increase apoptotic ability.

References

1 

Michaelis J: Osteosarcoma. Lancet. 1:11741988. View Article : Google Scholar : PubMed/NCBI

2 

Kansara M, Teng MW, Smyth MJ and Thomas DM: Translation biology of osteosarcoma. Nat Rev Cancer. 14:722–735. 2014. View Article : Google Scholar : PubMed/NCBI

3 

Gianferante DM, Mirabello L and Savage SA: Germline and somatic genetics of osteosarcoma-connecting aetiology, biology and therapy. Nat Rev Endocrinol. 13:480–491. 2017. View Article : Google Scholar : PubMed/NCBI

4 

Li ZX, Wang RY and Tang JC: Sodium valproate enhance γδ T cells killing osteosarcoma. Chin J Exp Surg. 33:154–157. 2016.

5 

Perron B, Lewit-Bentley A, Geny B and Russo-Marie F: Can enzymatic activity, or otherwise, be inferred from structural studies of annexin III? J Biol Chem. 272:11321–11326. 1997. View Article : Google Scholar : PubMed/NCBI

6 

Gerke V, Creutz CE and Moss SE: Annexins: Linking Ca2+ signalling to membrane dynamics. Nat Rev Mol Cell Biol. 6:449–461. 2005. View Article : Google Scholar : PubMed/NCBI

7 

Moss SE and Morgan RO: The annexins. Genome Biol. 5:2192004. View Article : Google Scholar : PubMed/NCBI

8 

Gerke V and Moss SE: Annexins: From structure to function. Physiol Rev. 82:331–371. 2002. View Article : Google Scholar : PubMed/NCBI

9 

Hamelin-Peyron C, Vlaeminck-Guillem V, Haidous H, Schwall GP, Poznanovic S, Gorius-Gallet E, Michles S, Larue A, Guillotte M, Ruffion A, et al: Prostate cancer biomarker annexin A3 detected in urines obtained following digital rectal examination presents antigenic variablility. Clin Biochem. 47:901–908. 2014. View Article : Google Scholar : PubMed/NCBI

10 

Wu N, Liu S, Guo C, Hou Z and Sun MZ: The role of annexin A3 playing in cancers. Clin Transl Oncol. 15:106–110. 2013. View Article : Google Scholar : PubMed/NCBI

11 

Zeidan B, Jackson TR, Larkin SE, Cutress RI, Coulton GR, Ashton-Key M, Murray N, Packham G, Gorgoulis V, Garbis SD and Townsend PA: Annexin A3 is a mammary marker and potential neoplastic breast cell therapeutic target. Oncotarget. 6:21421–21427. 2015. View Article : Google Scholar : PubMed/NCBI

12 

Tong M, Fung TM, Luk ST, Ng KY, Lee TK, Lin CH, Yam JW, Chan KW, Ng F, Zheng BJ, et al: ANXA3/JNK signaling promotes self-renewal and tumor growth, and its blockade provides a therapeutic target for hepatocellular carcinoma. Stem Cell Reports. 5:45–59. 2015. View Article : Google Scholar : PubMed/NCBI

13 

Liu YF, Xiao ZQ, Li MX, Zhang PF, Li C, Li F, Chen YH, Yi H, Yao HX and Chen ZC: Quantitative promote analysis reveals annexin A3 as a novel biomarker in lung adenocarcinoma. J Pathol. 217:54–64. 2009. View Article : Google Scholar : PubMed/NCBI

14 

Liu YF, Chen YH, Li MY, Zhang PF, Li GQ, Xiao ZQ and Chen ZC: Quantitative proteomic analysis identifying three annexins as lymph node metastasis-related proteins in lung adenocarcinoma. Med Oncol. 29:174–184. 2012. View Article : Google Scholar : PubMed/NCBI

15 

Pan QZ, Pan K, Weng DS, Zhao JJ, Zhang XF, Wang DD, Lv L, Jiang SS, Zheng HX and Xia JC: Annexin A3 promotes tumorigenesis and resistance to chemotherapy in hepatocellular carcinoma. Mol Carcinog. 54:598–607. 2015. View Article : Google Scholar : PubMed/NCBI

16 

Yan X, Yin J, Yao H, Mao N, Yang Y and Pan L: Increased expression of annexin A3 is a mechanism of platinum resistance in ovarian cancer. Cancer Res. 70:1616–1624. 2010. View Article : Google Scholar : PubMed/NCBI

17 

Tong SW, Yang YX, Hu HD, An X, Ye F, Hu P, Ren H, Li SL and Zhang DZ: Proteomic investigation of 5-flourouracil resistance in a human hepatocellular carcinoma cell line. J cell Biochem. 113:1671–1680. 2012.PubMed/NCBI

18 

Pénzváltó Z, Tegze B, Szász AM, Sztupinszki Z, Likó I, Szendrői A, Schäfer R and Győrffy B: Identifying resistance mechanisms against five tyrosine kinase inhibitors targeting the ERBB/RAS pathway in 45 cancer cell lines. PLoS One. 8:e595032013. View Article : Google Scholar : PubMed/NCBI

19 

Li ZX: Potential of human γδ T cells for immunotherapy of osteosarcoma. Mol Biol Rep. 40:427–437. 2013. View Article : Google Scholar : PubMed/NCBI

20 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI

21 

Mussunoor S and Murray GI: The role of annexins in tumor development and progression. J Pathol. 216:131–140. 2008. View Article : Google Scholar : PubMed/NCBI

22 

Shao P, Qu WK, Wang CY, Tian Y, Ye ML, Sun DG, Sui JD, Wang LM, Fan R and Gao ZM: MicroRNA-205-5p regulates the chemotherapeutic resistance of hepatocellular carcinoma cells by targeting PTEN/JNK/ANXA3 pathway. Am J Transl Res. 9:4300–4307. 2017.PubMed/NCBI

23 

Yang YM, Kim SY and Seki E: Inflammation and liver cancer: Molecular mechanisms and therapeutic targets. Semin Liver Dis. 39:26–42. 2019. View Article : Google Scholar : PubMed/NCBI

24 

Li Z, Zhang L, Gao M, Han M, Liu K, Zhang Z, Gong Z, Xing L, Shi X, Lu K and Gao H: Endoplasmic reticulum stress triggers xanthoangelol-induced protective autophagy via activation of JNK/c-Jun Axis in hepatocellular carcinoma. J Exp Clin Cancer Res. 38:82019. View Article : Google Scholar : PubMed/NCBI

25 

Kim JB, Lee S, Kim HR, Park SY, Lee M, Yoon JH and Kim YJ: Transforming growth factor-β decreases side population cells in hepatocellular carcinoma in vitro. Oncol Lett. 15:8723–8728. 2018.PubMed/NCBI

26 

Jin Y, Mao J, Wang H, Hou Z, Ma W, Zhang J, Wang B, Huang Y, Zang S, Tang J and Li L: Enhanced tumorigenesis and lymphatic metastasis of CD133+ hepatocarcinoma ascites syngeneic cell lines mediated by JNK signaling pathway in vitro and in vivo. Biomed Pharmacother. 67:337–345. 2013. View Article : Google Scholar : PubMed/NCBI

27 

Hagiwara S, Kudo M, Nagai T, Inoue T, Ueshima K, Nishida N, Watanabe T and Sakurai T: Activation of JNK and high expression level of CD133 predict a poor response to sorafenib in hepatocellular carcinoma. Br J Cancer. 106:1997–2003. 2012. View Article : Google Scholar : PubMed/NCBI

28 

Pan Q, Pan K, Wang QJ, Weng DS, Zhao JJ, Zheng HX, Zhang XF, Jiang SS, Lv L, Tang Y, et al: Annexin A3 as a potential target for immunotherapy of liver cancer stem-like cells. Stem Cells. 33:354–366. 2015. View Article : Google Scholar : PubMed/NCBI

29 

Zhou T, Li Y, Yang L, Yang L, Tang T, Zhang L and Shi J: Annexin A3 as a prognostic biomarker for breast cancer: A retrospective study. Biomed Res Int. 2017:26036852017. View Article : Google Scholar : PubMed/NCBI

30 

Zeng C, Ke Z, Song Y, Yao Y, Hu X, Zhang M, Li H and Yin J: Annexin A3 is associated with a poor prognosis in breast cancer and participates in the modulation of apoptosis in vitro by affecting the Bcl-2/Bax balance. Exp Mol Pathol. 95:23–31. 2013. View Article : Google Scholar : PubMed/NCBI

31 

Liu YF, Xiao ZQ, Li MX, Li MY, Zhang PF, Li C, Li F, Chen YH, Yi H, Yao HX and Chen ZC: Quantitative proteome analysis reveals annexin A3 as a novel biomarker in lung adenocarcinoma. J Pathol. 217:54–64. 2009. View Article : Google Scholar : PubMed/NCBI

32 

Li J, Zhou T, Liu L, Ju YC, Chen YT, Tan ZR and Wang J: The regulatory role of Annexin 3 in nude mouse bearing a subcutaneous xenograft of MDA-MB-231 human breast carcinoma. Pathol Res Pract. 214:1719–1725. 2018. View Article : Google Scholar : PubMed/NCBI

33 

Jeun M, Park S, Kim Y, Choi J, Song SH, Jeong IG, Kim CS and Lee KH: Self-Normalized detection of ANXA3 from untreated urine of prostate cancer patients without digital rectal examination. Adv Healthc Mater. 62017.

34 

Liu YF, Liu QQ, Zhang YH and Qiu JH: Annexin A3 knockdown suppresses lung adenocarcinoma. Anal Cell Pathol (Amst). 2016:41314032016.PubMed/NCBI

35 

Zhou T, Li Y, Yang L, Liu L, Ju Y and Li C: Silencing of ANXA3 expression by RNA interference inhibits the proliferation and invasion of breast cancer cells. Oncol Rep. 37:388–398. 2017. View Article : Google Scholar : PubMed/NCBI

36 

Li ZX, Wang RY and Tang JC: Type I IFN-mediated enhancement of anti-osteosarcoma cytotoxicity of human γδ T cells. Chin J Immun. 11:1533–1535, 1542. 2014.

37 

Li Z, Zhang J, Tang J and Wang R: Celastrol increases osteosarcoma cell lysis by γδ T cells through up-regulation of death receptors. Oncotarget. 7:84388–84397. 2016.PubMed/NCBI

38 

Li Z, Xu Q, Peng H, Cheng R, Sun Z and Ye Z: IFN-γ enhances HOS and U2OS cell lines susceptibility to γδ T cell-mediated killing through the Fas/Fas ligand pathway. Int Immunopharmacol. 11:496–503. 2011. View Article : Google Scholar : PubMed/NCBI

39 

Li Z, Peng H, Xu Q and Ye Z: Sensitization of human osteosarcoma cells to Vγ9Vδ2 T-cell-mediated cytotoxicity by zoledronate. J Orthop Res. 30:824–830. 2012. View Article : Google Scholar : PubMed/NCBI

40 

Li ZX, Zhang JZ, Wang ST, Wang RY and Tang JC: Celastrol increases osteosarcoma cells line HOS lysis by γδ T cells through TRAIL way. Chin J Immun. 32:1777–1780. 2016.

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September 2019
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Copy and paste a formatted citation
APA
Zeng, X., Wang, S., Gui, P., Wu, H., & Li, Z. (2019). Expression and significance of Annexin A3 in the osteosarcoma cell lines HOS and U2OS . Molecular Medicine Reports, 20, 2583-2590. https://doi.org/10.3892/mmr.2019.10513
MLA
Zeng, X., Wang, S., Gui, P., Wu, H., Li, Z."Expression and significance of Annexin A3 in the osteosarcoma cell lines HOS and U2OS ". Molecular Medicine Reports 20.3 (2019): 2583-2590.
Chicago
Zeng, X., Wang, S., Gui, P., Wu, H., Li, Z."Expression and significance of Annexin A3 in the osteosarcoma cell lines HOS and U2OS ". Molecular Medicine Reports 20, no. 3 (2019): 2583-2590. https://doi.org/10.3892/mmr.2019.10513