TRIM2 regulates the development and metastasis of tumorous cells of osteosarcoma

  • Authors:
    • Yi Qin
    • Jichao  Ye
    • Fulan Zhao
    • Shaoyu Hu
    • Suwei Wang
  • View Affiliations

  • Published online on: July 20, 2018     https://doi.org/10.3892/ijo.2018.4494
  • Pages: 1643-1656
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The present study aimed to investigate candidate genes involved in the development and metastasis of osteosarcoma. Candidate genes were screened preliminarily from the Gene Expression Omnibus database and then validated using actual tumor tissues collected from patients with osteosarcoma. The cells were prepared and transfected with specific gene-targeted small interfering RNA followed by an MTS assay for cell viability detection and Transwell assays for cell migration and invasion capacity detection. The cell apoptosis was determined by flow cytometry and the protein level of the genes was detected by western blot analysis. An in vivo nude model was used and injected with cells to detect the functions of the genes. Transcriptome sequencing was performed to verify the regulation network, followed by reverse transcription-quantitative polymerase chain reaction and western blot analyses for validation. Increased tripartite motif-containing protein 2 (TRIM2) was detected in the osteosarcoma tumor tissues compared with normal tissues. The inhibition of TRIM2 induced lower cell viability and cell invasion capacity, and increased the rate of cell apoptosis. Decreased TRIM2 also inhibited the development and metastasis of osteosarcoma in the nude mouse models. The transcriptome sequencing revealed that the regulation of TRIM2 may be correlated with genes, Sirtuin 4, DNA damage inducible transcript 3, cAMP responsive element binding protein 5, G protein-coupled receptor 65 (GPR65) and ADP-ribosyltransferase 5. Western blot analysis indicated that TRIM2 regulated the development and metastasis of osteosarcoma via the phosphoinositide 3-kinase/protein kinase B signaling pathway. Therefore, TRIM2 performs important functions in regulating the development and metastasis of osteosarcoma.

References

1 

Kashima T, Nakamura K, Kawaguchi J, Takanashi M, Ishida T, Aburatani H, Kudo A, Fukayama M and Grigoriadis AE: Overexpression of cadherins suppresses pulmonary metastasis of osteosarcoma in vivo. Int J Cancer. 104:147–154. 2003. View Article : Google Scholar : PubMed/NCBI

2 

Steeg PS: Tumor metastasis: Mechanistic insights and clinical challenges. Nat Med. 12:895–904. 2006. View Article : Google Scholar : PubMed/NCBI

3 

Bacci G, Rocca M, Salone M, Balladelli A, Ferrari S, Palmerini E, Forni C and Briccoli A: High grade osteosarcoma of the extremities with lung metastases at presentation: Treatment with neoadjuvant chemotherapy and simultaneous resection of primary and metastatic lesions. J Surg Oncol. 98:415–420. 2008. View Article : Google Scholar : PubMed/NCBI

4 

Harting MT, Blakely ML, Jaffe N, Cox CS Jr, Hayes-Jordan A, Benjamin RS, Raymond AK, Andrassy RJ and Lally KP: Long-term survival after aggressive resection of pulmonary metastases among children and adolescents with osteosarcoma. J Pediatr Surg. 41:194–199. 2006. View Article : Google Scholar : PubMed/NCBI

5 

Tsuchiya H, Kanazawa Y, Abdel-Wanis ME, Asada N, Abe S, Isu K, Sugita T and Tomita K: Effect of timing of pulmonary metastases identification on prognosis of patients with osteosarcoma: The Japanese Musculoskeletal Oncology Group study. J Clin Oncol. 20:3470–3477. 2002. View Article : Google Scholar : PubMed/NCBI

6 

Wu PK, Chen WM, Chen CF, Lee OK, Haung CK and Chen TH: Primary osteogenic sarcoma with pulmonary metastasis: Clinical results and prognostic factors in 91 patients. Jpn J Clin Oncol. 39:514–522. 2009. View Article : Google Scholar : PubMed/NCBI

7 

Tsuchiya H, Tomita K, Mori Y, Asada N and Yamamoto N: Marginal excision for osteosarcoma with caffeine assisted chemotherapy. Clin Orthop Relat Res. 358:27–35. 1999. View Article : Google Scholar

8 

Zanotti S and Canalis E: Notch signaling and the skeleton. Endocr Rev. 37:223–253. 2016. View Article : Google Scholar : PubMed/NCBI

9 

Kofler NM, Shawber CJ, Kangsamaksin T, Reed HO, Galatioto J and Kitajewski J: Notch signaling in developmental and tumor angiogenesis. Genes Cancer. 2:1106–1116. 2011. View Article : Google Scholar

10 

Engin F, Bertin T, Ma O, Jiang MM, Wang L, Sutton RE, Donehower LA and Lee B: Notch signaling contributes to the pathogenesis of human osteosarcomas. Hum Mol Genet. 18:1464–1470. 2009. View Article : Google Scholar : PubMed/NCBI

11 

Tanaka M, Setoguchi T, Hirotsu M, Gao H, Sasaki H, Matsunoshita Y and Komiya S: Inhibition of Notch pathway prevents osteosarcoma growth by cell cycle regulation. Br J Cancer. 100:1957–1965. 2009. View Article : Google Scholar : PubMed/NCBI

12 

Dailey DD, Anfinsen KP, Pfaff LE, Ehrhart EJ, Charles JB, Bønsdorff TB, Thamm DH, Powers BE, Jonasdottir TJ and Duval DL: HES1, a target of Notch signaling, is elevated in canine osteosarcoma, but reduced in the most aggressive tumors. BMC Vet Res. 9:1302013. View Article : Google Scholar : PubMed/NCBI

13 

Komori T: Regulation of osteoblast differentiation by transcription factors. J Cell Biochem. 99:1233–1239. 2006. View Article : Google Scholar : PubMed/NCBI

14 

Hilton MJ, Tu X, Wu X, Bai S, Zhao H, Kobayashi T, Kronenberg HM, Teitelbaum SL, Ross FP, Kopan R, et al: Notch signaling maintains bone marrow mesenchymal progenitors by suppressing osteoblast differentiation. Nat Med. 14:306–314. 2008. View Article : Google Scholar : PubMed/NCBI

15 

McManus MM, Weiss KR and Hughes DP: Understanding the role of Notch in osteosarcoma. Adv Exp Med Biol. 804:67–92. 2014. View Article : Google Scholar : PubMed/NCBI

16 

Krishnan V, Bryant HU and Macdougald OA: Regulation of bone mass by Wnt signaling. J Clin Invest. 116:1202–1209. 2006. View Article : Google Scholar : PubMed/NCBI

17 

Cai Y, Cai T and Chen Y: Wnt pathway in osteosarcoma, from oncogenic to therapeutic. J Cell Biochem 1. 15:625–631. 2014. View Article : Google Scholar

18 

Hoang BH, Kubo T, Healey JH, Sowers R, Mazza B, Yang R, Huvos AG, Meyers PA and Gorlick R: Expression of LDL receptor-related protein 5 (LRP5) as a novel marker for disease progression in high-grade osteosarcoma. Int J Cancer. 109:106–111. 2004. View Article : Google Scholar : PubMed/NCBI

19 

Ma Y, Ren Y, Han EQ, Li H, Chen D, Jacobs JJ, Gitelis S, O'Keefe RJ, Konttinen YT, Yin G, et al: Inhibition of the Wnt-β-catenin and Notch signaling pathways sensitizes osteosarcoma cells to chemotherapy. Biochem Biophys Res Commun. 431:274–279. 2013. View Article : Google Scholar : PubMed/NCBI

20 

Mödder UI, Oursler MJ, Khosla S and Monroe DG: Wnt10b activates the Wnt, notch, and NFκB pathways in U2OS osteosarcoma cells. J Cell Biochem. 112:1392–1402. 2011. View Article : Google Scholar

21 

Laplante M and Sabatini DM: mTOR signaling in growth control and disease. Cell. 149:274–293. 2012. View Article : Google Scholar : PubMed/NCBI

22 

Hu K, Dai HB and Qiu ZL: mTOR signaling in osteosarcoma: Oncogenesis and therapeutic aspects (Review). Oncol Rep. 36:1219–1225. 2016. View Article : Google Scholar : PubMed/NCBI

23 

Perry JA, Kiezun A, Tonzi P, Van Allen EM, Carter SL, Baca SC, Cowley GS, Bhatt AS, Rheinbay E, Pedamallu CS, et al: Complementary genomic approaches highlight the PI3K/mTOR pathway as a common vulnerability in osteosarcoma. Proc Natl Acad Sci USA. 111:E5564–E5573. 2014. View Article : Google Scholar : PubMed/NCBI

24 

Wang Z, Gerstein M and Snyder M: RNA-Seq: A revolutionary tool for transcriptomics. Nat Rev Genet. 10:57–63. 2009. View Article : Google Scholar

25 

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

26 

Thompson S, Pearson AN, Ashley MD, Jessick V, Murphy BM, Gafken P, Henshall DC, Morris KT, Simon RP and Meller R: Identification of a novel Bcl-2-interacting mediator of cell death (Bim) E3 ligase, tripartite motif-containing protein 2 (TRIM2), and its role in rapid ischemic tolerance-induced neuroprotection. J Biol Chem. 286:19331–19339. 2011. View Article : Google Scholar : PubMed/NCBI

27 

Chen X, Dong C, Law PT, Chan MT, Su Z, Wang S, Wu WK and Xu H: MicroRNA-145 targets TRIM2 and exerts tumor-suppressing functions in epithelial ovarian cancer. Gynecol Oncol. 139:513–519. 2015. View Article : Google Scholar : PubMed/NCBI

28 

Williams MD, Zhang L, Elliott DD, Perrier ND, Lozano G, Clayman GL and El-Naggar AK: Differential gene expression profiling of aggressive and nonaggressive follicular carcinomas. Hum Pathol. 42:1213–1220. 2011. View Article : Google Scholar : PubMed/NCBI

29 

Panaccione A, Guo Y, Yarbrough WG and Ivanov SV: Expression profiling of clinical specimens supports the existence of neural progenitor-like stem cells in basal breast cancers. Clin Breast Cancer. 17:298–306.e7. 2017. View Article : Google Scholar : PubMed/NCBI

30 

Ivanov SV, Panaccione A, Nonaka D, Prasad ML, Boyd KL, Brown B, Guo Y, Sewell A and Yarbrough WG: Diagnostic SOX10 gene signatures in salivary adenoid cystic and breast basal-like carcinomas. Br J Cancer. 109:444–451. 2013. View Article : Google Scholar : PubMed/NCBI

31 

Yang K, Zhang G, Mei J, Chen D and Wu M: Screening and analysis of pathogenic genes during DMBA-induced buccal mucosa carcinogenesis in golden hamsters. Oncol Rep. 23:1619–1624. 2010. View Article : Google Scholar : PubMed/NCBI

32 

Miyatake T, Ueda Y, Nakashima R, Yoshino K, Kimura T, Murata T, Nomura T, Fujita M, Buzard GS and Enomoto T: Down-regulation of insulin-like growth factor binding protein-5 (IGFBP-5): Novel marker for cervical carcinogenesis. Int J Cancer. 120:2068–2077. 2007. View Article : Google Scholar : PubMed/NCBI

33 

Thiery JP, Acloque H, Huang RY and Nieto MA: Epithelial-mesenchymal transitions in development and disease. Cell. 139:871–890. 2009. View Article : Google Scholar : PubMed/NCBI

34 

Willis BC, Liebler JM, Luby-Phelps K, Nicholson AG, Crandall ED, du Bois RM and Borok Z: Induction of epithelial-mesenchymal transition in alveolar epithelial cells by transforming growth factor-beta1: Potential role in idiopathic pulmonary fibrosis. Am J Pathol. 166:1321–1332. 2005. View Article : Google Scholar : PubMed/NCBI

35 

Willis BC and Borok Z: TGF-beta-induced EMT: Mechanisms and implications for fibrotic lung disease. Am J Physiol Lung Cell Mol Physiol. 293:L525–L534. 2007. View Article : Google Scholar : PubMed/NCBI

36 

Moses MA, Wiederschain D, Loughlin KR, Zurakowski D, Lamb CC and Freeman MR: Increased incidence of matrix metalloproteinases in urine of cancer patients. Cancer Res. 58:1395–1399. 1998.PubMed/NCBI

37 

Finkel T, Deng CX and Mostoslavsky R: Recent progress in the biology and physiology of sirtuins. Nature. 460:587–591. 2009. View Article : Google Scholar : PubMed/NCBI

38 

Haigis MC and Guarente LP: Mammalian sirtuins–emerging roles in physiology, aging, and calorie restriction. Genes Dev. 20:2913–2921. 2006. View Article : Google Scholar : PubMed/NCBI

39 

Jeong SM, Xiao C, Finley LW, Lahusen T, Souza AL, Pierce K, Li YH, Wang X, Laurent G, German NJ, et al: SIRT4 has tumor-suppressive activity and regulates the cellular metabolic response to DNA damage by inhibiting mitochondrial glutamine metabolism. Cancer Cell. 23:450–463. 2013. View Article : Google Scholar : PubMed/NCBI

40 

Shi Q, Liu T, Zhang X, Geng J, He X, Nu M and Pang D: Decreased sirtuin 4 expression is associated with poor prognosis in patients with invasive breast cancer. Oncol Lett. 12:2606–2612. 2016. View Article : Google Scholar : PubMed/NCBI

41 

Qi L and Ding Y: Involvement of the CREB5 regulatory network in colorectal cancer metastasis. Yi Chuan. 36:679–684. 2014.PubMed/NCBI

42 

Harada Y, Tamura Y and Endo T: Identification of yeast Art5 as a multicopy suppressor for the mitochondrial translocator maintenance protein Tam41. Biochem Biophys Res Commun. 392:228–233. 2010. View Article : Google Scholar : PubMed/NCBI

43 

Shih YL, Chou HM, Chou HC, Lu HF, Chu YL, Shang HS and Chung JG: Casticin impairs cell migration and invasion of mouse melanoma B16F10 cells via PI3K/AKT and NF-κB signaling pathways. Environ Toxicol. 32:2097–2112. 2017. View Article : Google Scholar : PubMed/NCBI

44 

Wang J, Wang L, Ho CT, Zhang K, Liu Q and Zhao H: Garcinol from Garcinia indica downregulates cancer stem-like cell biomarker ALDH1A1 in nonsmall cell lung cancer A549 cells through DDIT3 activation. J Agric Food Chem. 65:3675–3683. 2017. View Article : Google Scholar : PubMed/NCBI

45 

Čokić VP, Smith RD, Biancotto A, Noguchi CT, Puri RK and Schechter AN: Globin gene expression in correlation with G protein-related genes during erythroid differentiation. BMC Genomics. 14:1162013. View Article : Google Scholar : PubMed/NCBI

46 

Lian Z, Han J, Huang L, Wei C, Fan Y, Xu J, Zhou M, Feng H, Liu Q, Chen L, et al: A005, a novel inhibitor of phosphatidylinositol 3-kinase/mammalian target of rapamycin, prevents osteosarcoma-induced osteolysis. Carcinogenesis. 2018. View Article : Google Scholar : PubMed/NCBI

47 

Yu G, Liu G, Yuan D, Dai J, Cui Y and Tang X: Long non-coding RNA ANRIL is associated with a poor prognosis of osteosarcoma and promotes tumorigenesis via PI3K/Akt pathway. J Bone Oncol. 11:51–55. 2018. View Article : Google Scholar : PubMed/NCBI

48 

Chen L, Pei H, Lu SJ, Liu ZJ, Yan L, Zhao XM, Hu B and Lu HG: SPOP suppresses osteosarcoma invasion via PI3K/AKT/NF-κB signaling pathway. Eur Rev Med Pharmacol Sci. 22:609–615. 2018.PubMed/NCBI

49 

Hu B, Lv X, Gao F, Chen S, Wang S, Qing X, Liu J, Wang B and Shao Z: Downregulation of DEPTOR inhibits the proliferation, migration, and survival of osteosarcoma through PI3K/Akt/mTOR pathway. OncoTargets Ther. 10:4379–4391. 2017. View Article : Google Scholar

50 

Pu Y, Yi Q, Zhao F, Wang H, Cai W and Cai S: MiR-20a-5p represses multi-drug resistance in osteosarcoma by targeting the KIF26B gene. Cancer Cell Int. 16:642016. View Article : Google Scholar : PubMed/NCBI

51 

Fujimori A, Cheng SL, Avioli LV and Civitelli R: Structure-function relationship of parathyroid hormone: Activation of phospholipase-C, protein kinase-A and -C in osteosarcoma cells. Endocrinology. 130:29–36. 1992. View Article : Google Scholar : PubMed/NCBI

52 

Miles RR, Sluka JP, Halladay DL, Santerre RF, Hale LV, Bloem L, Thirunavukkarasu K, Galvin RJ, Hock JM and Onyia JE: ADAMTS-1: A cellular disintegrin and metalloprotease with thrombospondin motifs is a target for parathyroid hormone in bone. Endocrinology. 141:4533–4542. 2000. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

October 2018
Volume 53 Issue 4

Print ISSN: 1019-6439
Online ISSN:1791-2423

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Qin, Y., Ye, J., Zhao, F., Hu, S., & Wang, S. (2018). TRIM2 regulates the development and metastasis of tumorous cells of osteosarcoma. International Journal of Oncology, 53, 1643-1656. https://doi.org/10.3892/ijo.2018.4494
MLA
Qin, Y., Ye, J., Zhao, F., Hu, S., Wang, S."TRIM2 regulates the development and metastasis of tumorous cells of osteosarcoma". International Journal of Oncology 53.4 (2018): 1643-1656.
Chicago
Qin, Y., Ye, J., Zhao, F., Hu, S., Wang, S."TRIM2 regulates the development and metastasis of tumorous cells of osteosarcoma". International Journal of Oncology 53, no. 4 (2018): 1643-1656. https://doi.org/10.3892/ijo.2018.4494