Advertisement

Tumor Biology

, Volume 36, Issue 4, pp 2885–2892 | Cite as

Overexpression of Numb suppresses growth, migration, and invasion of human clear cell renal cell carcinoma cells

  • Jin Sima
  • Bao Zhang
  • Yuanzi Yu
  • Xinyuan Sima
  • Yanxin Mao
Research Article

Abstract

The objective of the study was to investigate the impact of Numb on cell growth, cell migration, and invasion in human clear cell renal cell carcinoma (ccRCC). Endogenous expression of Numb was evaluated in the ccRCC cell lines (786-O, Caki-1, and Caki-2) and control reference human renal proximal tubular epithelial cells. Numb expression was decreased in the ccRCC cells compared with the control cells (P < 0.01). Then, 786-O and Caki-1 cells described as suitable transfection hosts were used in transfection to carry out biological function studies. The three experimental groups were as follows: Numb-ORF (transfected with Numb-ORF plasmid), blank-vector (transfected with pCMV6-entry), and cell-alone group (no DNA). Numb expression in the Numb-ORF groups was significantly higher than that in the controls (P < 0.01). Cell growth was remarkably reduced (P < 0.01), and the number of migrating or invading cells was reduced (P < 0.01) in the Numb-ORF groups compared with controls. Furthermore, the ratio of G0/G1 phase in the Numb-ORF group of 786-O cells was increased, and the S phase fraction and proliferation index was decreased (P < 0.01). Cyclin D1 and MMP-9 expression was reduced in the Numb-ORF groups compared with controls. Here, we have provided data for attenuated Numb expression in the ccRCC cells. Overexpression of Numb could induce G0/G1 phase arrest and inhibit cell proliferation, migration, and invasion. The suppressive effects might be due to downregulation of cyclin D1 or MMP-9 expression. Taken together, our data suggest that Numb may possibly function as a tumor suppressor involved in the carcinogenesis of ccRCC.

Keywords

Numb Clear cell renal cell carcinoma Proliferation Migration Invasion Cyclin D1 MMP-9 

Notes

Conflicts of interest

None

References

  1. 1.
    Ai Q, Ma X, Huang Q, Liu S, Shi T, Zhang C, et al. High-level expression of Notch1 increased the risk of metastasis in T1 stage clear cell renal cell carcinoma. PLoS One. 2012;7(4):e35022.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Liu S, Ma X, Ai Q, Huang Q, Shi T, Zhu M, et al. NOTCH1 functions as an oncogene by regulating the PTEN/PI3K/AKT pathway in clear cell renal cell carcinoma. Urol Oncol. 2013;31(6):938–48.CrossRefPubMedGoogle Scholar
  3. 3.
    Wei C, Wu S, Li X, Wang Y, Ren R, Lai Y, et al. High expression of FER tyrosine kinase predicts poor prognosis in clear cell renal cell carcinoma. Oncol Lett. 2013;5(2):473–8.PubMedGoogle Scholar
  4. 4.
    Ma X, Fan Y, Gao Y, Zhang Y, Huang Q, Ai Q, et al. Dicer is down-regulated in clear cell renal cell carcinoma and in vitro Dicer knockdown enhances malignant phenotype transformation. Urol Oncol. 2014;32(1):e9–17.CrossRefGoogle Scholar
  5. 5.
    Song E, Ma X, Li H, Zhang P, Ni D, Chen W, et al. Attenuation of krüppel-like factor 4 facilitates carcinogenesis by inducing G1/S phase arrest in clear cell renal cell carcinoma. PLoS One. 2013;8(7):e67758.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Rini BI, Campbell SC, Escudier B. Renal cell carcinoma. Lancet. 2009;373(9669):1119–32.CrossRefPubMedGoogle Scholar
  7. 7.
    Castellano D, Virizuela J, Cruz J, Sepulveda J, Saez M, Paz-Ares L. The role of pharmacogenomics in metastatic renal cell carcinoma. Cancer Metastasis Rev. 2012;31(S1):S29–32.CrossRefPubMedGoogle Scholar
  8. 8.
    Ma X, Gao Y, Fan Y, Ni D, Zhang Y, Chen W, et al. Overexpression of E2F1 promotes tumor malignancy and correlates with TNM stages in clear cell renal cell carcinoma. PLoS One. 2013;8(9):e73436.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Vogelzang NJ. Treatment options in metastatic renal carcinoma: an embarrassment of riches. J Clin Oncol. 2006;24(1):1–3.CrossRefPubMedGoogle Scholar
  10. 10.
    Wei S, Hao C, Li X, Zhao H, Chen J, Zhou Q. Effects of BTG2 on proliferation inhibition and anti-invasion in human lung cancer cells. Tumour Biol. 2012;33(4):1223–30.CrossRefPubMedGoogle Scholar
  11. 11.
    Zoratto F, Rossi L, Verrico M, Papa A, Basso E, Zullo A, et al. Focus on genetic and epigenetic events of colorectal cancer pathogenesis: implications for molecular diagnosis. Tumour Biol. 2014;35(7):6195–206.CrossRefPubMedGoogle Scholar
  12. 12.
    Hu F, Meng X, Tong Q, Liang L, Xiang R, Zhu T, et al. BMP-6 inhibits cell proliferation by targeting microRNA-192 in breast cancer. Biochim Biophys Acta. 2013;1832(12):2379–90.CrossRefPubMedGoogle Scholar
  13. 13.
    Yu Y, Yin D, Hoque MO, Cao B, Jia Y, Yang Y, et al. AKT signaling pathway activated by HIN-1 methylation in non-small cell lung cancer. Tumour Biol. 2012;33(2):307–14.CrossRefPubMedGoogle Scholar
  14. 14.
    Li Q, Shi R, Wang Y, Niu X. TAGLN suppresses proliferation and invasion, and induces apoptosis of colorectal carcinoma cells. Tumour Biol. 2013;34(1):505–13.CrossRefPubMedGoogle Scholar
  15. 15.
    Li H, Zhang Y, Zhang Y, Bai X, Peng Y, He P. TRIM31 is downregulated in non-small cell lung cancer and serves as a potential tumor suppressor. Tumour Biol. 2014;35(6):5747–52.CrossRefPubMedGoogle Scholar
  16. 16.
    Zheng Y, Wang X, Wang H, Yan W, Zhang Q, Chang X. Bone morphogenetic protein 2 inhibits hepatocellular carcinoma growth and migration through downregulation of the PI3K/AKT pathway. Tumour Biol. 2014;35(6):5189–98.CrossRefPubMedGoogle Scholar
  17. 17.
    Pedersen WA, Chan SL, Zhu H, Abdur-Rahman LA, Verdi JM, Mattson MP. Numb isoforms containing a short PTB domain promote neurotrophic factor-induced differentiation and neurotrophic factor withdrawal-induced death of PC12 cells. J Neurochem. 2002;82(4):976–86.CrossRefPubMedGoogle Scholar
  18. 18.
    Colaluca IN, Tosoni D, Nuciforo P, Senic-Matuglia F, Galimberti V, Viale G, et al. NUMB controls p53 tumour suppressor activity. Nature. 2008;451(7174):76–80.CrossRefPubMedGoogle Scholar
  19. 19.
    Carter S, Vousden KH. A role for Numb in p53 stabilization. Genome Biol. 2008;9(5):221–3.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Pece S, Serresi M, Santolini E, Capra M, Hulleman E, Galimberti V, et al. Loss of negative regulation by Numb over Notch is relevant to human breast carcinogenesis. J Cell Biol. 2004;167(2):215–21.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Gulino A, Di Marcotullio L, Screpanti I. The multiple functions of Numb. Exp Cell Res. 2010;316(6):900–6.CrossRefPubMedGoogle Scholar
  22. 22.
    Thol F, Winschel C, Sonntag AK, Damm F, Wagner K, Chaturvedi A, et al. Prognostic significance of expression levels of stem cell regulators MSI2 and NUMB in acute myeloid leukemia. Ann Hematol. 2013;92(3):315–23.CrossRefPubMedGoogle Scholar
  23. 23.
    Westhoff B, Colaluca IN, DArio G, Donzelli M, Tosoni D, Volorio S, et al. Alterations of the Notch pathway in lung cancer. Proc Natl Acad Sci U S A. 2009;106(52):22293–8.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Maiorano E, Favia G, Pece S, Resta L, Maisonneuve P, Di Fiore PP, et al. Prognostic implications of NUMB immunoreactivity in salivary gland carcinomas. Int J Immunopathol Pharmacol. 2007;20(4):779–89.CrossRefPubMedGoogle Scholar
  25. 25.
    Yan B, Omar FM, Das K, Ng WH, Lim C, Shiuan K, et al. Characterization of Numb expression in astrocytomas. Neuropathology. 2008;28(5):479–84.CrossRefPubMedGoogle Scholar
  26. 26.
    Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method. Nat Protoc. 2008;3(6):1101–8.CrossRefPubMedGoogle Scholar
  27. 27.
    White NM, Newsted DW, Masui O, Romaschin AD, Siu KW, Yousef GM. Identification and validation of dysregulated metabolic pathways in metastatic renal cell carcinoma. Tumour Biol. 2014;35(3):1833–46.CrossRefPubMedGoogle Scholar
  28. 28.
    Escudier B, Eisen T, Stadler WM, Szczylik C, Oudard S, Siebels M, et al. Sorafenib in advanced clear-cell renal-cell carcinoma. N Engl J Med. 2007;356(2):125–34.CrossRefPubMedGoogle Scholar
  29. 29.
    Motzer RJ, Hutson TE, Cella D, Reeves J, Hawkins R, Guo J, et al. Pazopanib versus sunitinib in metastatic renal-cell carcinoma. N Engl J Med. 2013;369(8):722–31.CrossRefPubMedGoogle Scholar
  30. 30.
    Rennstam K, McMichael N, Berglund P, Honeth G, Hegardt C, Ryden L, et al. Numb protein expression correlates with a basal-like phenotype and cancer stem cell markers in primary breast cancer. Breast Cancer Res Treat. 2010;122(2):315–24.CrossRefPubMedGoogle Scholar
  31. 31.
    Tang ZP, Cui QZ, Dong QZ, Xu K, Wang EH. Ataxia-telangiectasia group D complementing gene (ATDC) upregulates matrix metalloproteinase 9 (MMP-9) to promote lung cancer cell invasion by activating ERK and JNK pathways. Tumour Biol. 2013;34(5):2835–42.CrossRefPubMedGoogle Scholar
  32. 32.
    Sun GG, Lu YF, Zhang J, Hu WN. Filamin A regulates MMP-9 expression and suppresses prostate cancer cell migration and invasion. Tumour Biol. 2014;35(4):3819–26.CrossRefPubMedGoogle Scholar
  33. 33.
    Sato A, Nagase H, Obinata D, Fujiwara K, Fukuda N, Soma M, et al. Inhibition of MMP-9 using a pyrrole-imidazole polyamide reduces cell invasion in renal cell carcinoma. Int J Oncol. 2013;43(5):1441–6.PubMedGoogle Scholar
  34. 34.
    Hu Z, Zhou Z, Xiong G, Wang Y, Lai Y, Deng L, et al. Cyclin D1 G870A polymorphism and the risk of hepatocellular carcinoma in a Chinese population. Tumour Biol. 2014;35(6):5607–12.CrossRefPubMedGoogle Scholar
  35. 35.
    Lima MS, Pereira RA, Costa RS, Tucci S, Dantas M, Muglia VF, et al. The prognostic value of cyclin D1 in renal cell carcinoma. Int Urol Nephrol. 2014;46(5):905–13.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Jin Sima
    • 1
    • 2
  • Bao Zhang
    • 1
    • 2
  • Yuanzi Yu
    • 3
  • Xinyuan Sima
    • 4
  • Yanxin Mao
    • 5
  1. 1.Department of UrologyAerospace Central HospitalBeijingChina
  2. 2.Key Laboratory of Oncogenesis and Related Genes ResearchAerospace Clinical Medical College of Peking UniversityBeijingChina
  3. 3.Department of GastroenterologyProvincial Hospital Affiliated to Shandong UniversityJinanChina
  4. 4.Public Health Care CenterGeneral Hospital of Beijing Military AreaBeijingChina
  5. 5.Department of PharmacyJingxi Hospital of Beijing Military AreaBeijingChina

Personalised recommendations