Advertisement

Effect of RAB31 silencing on osteosarcoma cell proliferation and migration through the Hedgehog signaling pathway

  • Qiong Yu
  • Dong Li
  • Dan Wang
  • Chun-Mei Hu
  • Yan Sun
  • Yan Tang
  • Guang ShiEmail author
Original Article
  • 60 Downloads

Abstract

Osteosarcoma (OS) is a prevalent cancer that plagues people worldwide. Identifying prognostic markers would be useful in treating human OS. In this study, we aimed to explore the functions of Ras-related protein Rab-31 (RAB31) in OS-cell proliferation, migration, and invasion as well as its roles in the Hedgehog signaling pathway for better understanding of the mechanism. To assess the detailed regulatory mechanism of RAB31 silencing on OS, both RT-qPCR and Western blot analysis were employed to evaluate the expressions of RAB31 as well as the Hedgehog signaling pathway-related genes. Besides, we also investigated the effects of silenced RAB31 both in vitro and in vivo. First, we found that in OS tissues, both mRNA and protein expressions of RAB31 and PCNA had a significant increase. Second, the Hedgehog signaling pathway was detected to play an integral role in OS progression. Finally, after transfection of RAB31-siRNA to reduce the expression of RAB31, the Hedgehog signaling pathway was suppressed, along with cell proliferation, invasion, and migration. Therefore, we conclude that RAB31 plays an important role in OS development and its silencing delays the OS progression via suppression of the Hedgehog signaling pathway.

Keywords

RAB31 Osteosarcoma Hedgehog signaling pathway Proliferation Invasion 

Notes

Acknowledgements

We thank the reviewers for critical comments.

Compliance with ethical standards

Conflicts of interest

The authors declares that they have conflict of interest.

Ethical approval

All aspects of the study are in strict accordance with the Declaration of Helsinki. All of the above specimens were collected with informed consent of patients and all the patients signed informed consent.

References

  1. 1.
    Liu T, Zhou W, Zhang F, Shi G, Teng H, Xiao J, Wang Y (2014) Knockdown of IRX2 inhibits osteosarcoma cell proliferation and invasion by the AKT/MMP9 signaling pathway. Mol Med Rep 10:169–174CrossRefGoogle Scholar
  2. 2.
    Luetke A, Meyers PA, Lewis I, Juergens H (2014) Osteosarcoma treatment—where do we stand? A state of the art review. Cancer Treat Rev 40:523–532CrossRefGoogle Scholar
  3. 3.
    Savage SA, Mirabello L (2011) Using epidemiology and genomics to understand osteosarcoma etiology. Sarcoma 2011:548151CrossRefGoogle Scholar
  4. 4.
    Huang J, Ni J, Liu K, Yu Y, Xie M, Kang R, Vernon P, Cao L, Tang D (2012) HMGB1 promotes drug resistance in osteosarcoma. Cancer Res 72:230–238CrossRefGoogle Scholar
  5. 5.
    Mirabello L, Pfeiffer R, Murphy G, Daw NC, Patino-Garcia A, Troisi RJ, Hoover RN, Douglass C, Schuz J, Craft AW, Savage SA (2011) Height at diagnosis and birth-weight as risk factors for osteosarcoma. Cancer Causes Control 22:899–908CrossRefGoogle Scholar
  6. 6.
    Wang Y, Li L, Shao N, Hu Z, Chen H, Xu L, Wang C, Cheng Y, Xiao J (2015) Triazine-modified dendrimer for efficient TRAIL gene therapy in osteosarcoma. Acta Biomater 17:115–124CrossRefGoogle Scholar
  7. 7.
    Bienemann K, Staege MS, Howe SJ, Sena-Esteves M, Hanenberg H, Kramm CM (2013) Targeted expression of human folylpolyglutamate synthase for selective enhancement of methotrexate chemotherapy in osteosarcoma cells. Cancer Gene Ther 20:514–520CrossRefGoogle Scholar
  8. 8.
    Xu M, Xu SF, Yu XC (2014) Clinical analysis of osteosarcoma patients treated with high-dose methotrexate-free neoadjuvant chemotherapy. Curr Oncol 21:e678–684CrossRefGoogle Scholar
  9. 9.
    Oertel S, Blattmann C, Rieken S, Jensen A, Combs SE, Huber PE, Bischof M, Kulozik A, Debus J, Schulz-Ertner D (2010) Radiotherapy in the treatment of primary osteosarcoma—a single center experience. Tumori J 96:582–588CrossRefGoogle Scholar
  10. 10.
    Chua CE, Tang BL (2015) The role of the small GTPase Rab31 in cancer. J Cell Mol Med 19:1–10CrossRefGoogle Scholar
  11. 11.
    Chua CE, Tang BL (2014) Engagement of the small GTPase Rab31 protein and its effector, early endosome antigen 1, is important for trafficking of the ligand-bound epidermal growth factor receptor from the early to the late endosome. J Biol Chem 289:12375–12389CrossRefGoogle Scholar
  12. 12.
    Grismayer B, Solch S, Seubert B, Kirchner T, Schafer S, Baretton G, Schmitt M, Luther T, Kruger A, Kotzsch M, Magdolen V (2012) Rab31 expression levels modulate tumor-relevant characteristics of breast cancer cells. Mol Cancer 11:62CrossRefGoogle Scholar
  13. 13.
    Jin C, Rajabi H, Pitroda S, Li A, Kharbanda A, Weichselbaum R, Kufe D (2012) Cooperative interaction between the MUC1-C oncoprotein and the Rab31 GTPase in estrogen receptor-positive breast cancer cells. PLoS One 7:e39432CrossRefGoogle Scholar
  14. 14.
    Serao NV, Delfino KR, Southey BR, Beever JE, Rodriguez-Zas SL (2011) Cell cycle and aging, morphogenesis, and response to stimuli genes are individualized biomarkers of glioblastoma progression and survival. BMC Med Genom 4:49CrossRefGoogle Scholar
  15. 15.
    Chan LH, Wang W, Yeung W, Deng Y, Yuan P, Mak KK (2014) Hedgehog signaling induces osteosarcoma development through Yap1 and H19 overexpression. Oncogene 33:4857–4866CrossRefGoogle Scholar
  16. 16.
    Briscoe J, Therond PP (2013) The mechanisms of Hedgehog signalling and its roles in development and disease. Nat Rev Mol Cell Biol 14:416–429CrossRefGoogle Scholar
  17. 17.
    Pan Y, Zhang Y, Chen L, Liu Y, Feng Y, Yan J (2016) The Critical Role of Rab31 in Cell Proliferation and Apoptosis in Cancer Progression. Mol Neurobiol 53:4431–4437CrossRefGoogle Scholar
  18. 18.
    Cao CM, Yang FX, Wang PL, Yang QX, Sun XR (2014) Clinicopathologic significance of S100A4 expression in osteosarcoma. Eur Rev Med Pharmacol Sci 18:833–839PubMedGoogle Scholar
  19. 19.
    Enneking WF, Spanier SS, Goodman MA (2003) A system for the surgical staging of musculoskeletal sarcoma. 1980. Clin Orthop Relat Res 4–18.  https://doi.org/10.1097/01.blo.0000093891.12372.0f CrossRefGoogle Scholar
  20. 20.
    Vergara-Lluri ME, Moatamed NA, Hong E, Apple SK (2012) High concordance between HercepTest immunohistochemistry and ERBB2 fluorescence in situ hybridization before and after implementation of American Society of Clinical Oncology/College of American Pathology 2007 guidelines. Mod Pathol 25:1326–1332CrossRefGoogle Scholar
  21. 21.
    Warzecha J, Gottig S, Chow KU, Bruning C, Percic D, Boehrer S, Brude E, Kurth A (2007) Inhibition of osteosarcoma cell proliferation by the Hedgehog-inhibitor cyclopamine. J Chemother 19:554–561CrossRefGoogle Scholar
  22. 22.
    Li S, Sun W, Wang H, Zuo D, Hua Y, Cai Z (2015) Research progress on the multidrug resistance mechanisms of osteosarcoma chemotherapy and reversal. Tumour Biol 36:1329–1338CrossRefGoogle Scholar
  23. 23.
    Yamamoto N, Tsuchiya H (2013) Chemotherapy for osteosarcoma—where does it come from? What is it? Where is it going? Expert Opin Pharmacother 14:2183–2193CrossRefGoogle Scholar
  24. 24.
    Yang D, Liu G, Wang K (2015) miR-203 acts as a tumor suppressor gene in osteosarcoma by regulating RAB22A. PLoS One 10:e0132225CrossRefGoogle Scholar
  25. 25.
    Shahi MH, Holt R, Rebhun RB (2014) Blocking signaling at the level of GLI regulates downstream gene expression and inhibits proliferation of canine osteosarcoma cells. PLoS One 9:e96593CrossRefGoogle Scholar
  26. 26.
    Hirotsu M, Setoguchi T, Sasaki H, Matsunoshita Y, Gao H, Nagao H, Kunigou O, Komiya S (2010) Smoothened as a new therapeutic target for human osteosarcoma. Mol Cancer 9:5CrossRefGoogle Scholar
  27. 27.
    Wang W, Luo H, Wang A (2006) Expression of survivin and correlation with PCNA in osteosarcoma. J Surg Oncol 93:578–584CrossRefGoogle Scholar
  28. 28.
    Piro F, Leonardi L (2015) Expression of Bcl-2 in canine osteosarcoma. Open Vet J 5:27–29PubMedPubMedCentralGoogle Scholar
  29. 29.
    Wang D, Bi Z (2014) Bufalin inhibited the growth of human osteosarcoma MG-63 cells via down-regulation of Bcl-2/Bax and triggering of the mitochondrial pathway. Tumour Biol 35:4885–4890CrossRefGoogle Scholar
  30. 30.
    Wuichet K, Sogaard-Andersen L (2014) Evolution and diversity of the Ras superfamily of small GTPases in prokaryotes. Genome Biol Evol 7:57–70CrossRefGoogle Scholar
  31. 31.
    Warzecha J, Dinges D, Kaszap B, Henrich D, Marzi I, Seebach C (2012) Effect of the Hedgehog-inhibitor cyclopamine on mice with osteosarcoma pulmonary metastases. Int J Mol Med 29:423–427PubMedGoogle Scholar
  32. 32.
    Paget C, Duret H, Ngiow SF, Kansara M, Thomas DM, Smyth MJ (2012) Studying the role of the immune system on the antitumor activity of a Hedgehog inhibitor against murine osteosarcoma. Oncoimmunology 1:1313–1322CrossRefGoogle Scholar
  33. 33.
    Saitoh Y, Setoguchi T, Nagata M, Tsuru A, Nakamura S, Nagano S, Ishidou Y, Nagao-Kitamoto H, Yokouchi M, Maeda S, Tanimoto A, Furukawa T, Komiya S (2016) Combination of Hedgehog inhibitors and standard anticancer agents synergistically prevent osteosarcoma growth. Int J Oncol 48:235–242CrossRefGoogle Scholar
  34. 34.
    Ding YL, Zhou Y, Xiang L, Ji ZP, Luo ZH (2012) Expression of glioma-associated oncogene homolog 1 is associated with invasion and postoperative liver metastasis in colon cancer. Int J Med Sci 9:334–338CrossRefGoogle Scholar
  35. 35.
    Tao Y, Mao J, Zhang Q, Li L (2011) Overexpression of Hedgehog signaling molecules and its involvement in triple-negative breast cancer. Oncol Lett 2:995–1001PubMedPubMedCentralGoogle Scholar
  36. 36.
    You S, Zhou J, Chen S, Zhou P, Lv J, Han X, Sun Y (2010) PTCH1, a receptor of Hedgehog signaling pathway, is correlated with metastatic potential of colorectal cancer. Ups J Med Sci 115:169–175CrossRefGoogle Scholar
  37. 37.
    Sui Y, Zheng X, Zhao D (2015) Rab31 promoted hepatocellular carcinoma (HCC) progression via inhibition of cell apoptosis induced by PI3K/AKT/Bcl-2/BAX pathway. Tumour Biol 36:8661–8670CrossRefGoogle Scholar
  38. 38.
    Lo WW, Wunder JS, Dickson BC, Campbell V, McGovern K, Alman BA, Andrulis IL (2014) Involvement and targeted intervention of dysregulated Hedgehog signaling in osteosarcoma. Cancer 120:537–547CrossRefGoogle Scholar
  39. 39.
    Eggenschwiler JT, Espinoza E, Anderson KV (2001) Rab23 is an essential negative regulator of the mouse Sonic hedgehog signalling pathway. Nature 412:194–198CrossRefGoogle Scholar

Copyright information

© The Japanese Society for Bone and Mineral Research and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Qiong Yu
    • 1
  • Dong Li
    • 2
  • Dan Wang
    • 3
  • Chun-Mei Hu
    • 1
  • Yan Sun
    • 1
  • Yan Tang
    • 1
  • Guang Shi
    • 1
    Email author
  1. 1.Department of Hematology and OncologyThe Second Hospital of Jilin UniversityChangchunPeople’s Republic of China
  2. 2.Department of Obstetrics and GynecologyThe Second Hospital of Jilin UniversityChangchunPeople’s Republic of China
  3. 3.Department of Breast SurgeryThe Second Hospital of Jilin UniversityChangchunPeople’s Republic of China

Personalised recommendations