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Tumor Biology

, Volume 36, Issue 4, pp 2393–2401 | Cite as

miR-194 targets RBX1 gene to modulate proliferation and migration of gastric cancer cells

  • Xiaonan Chen
  • Yuanyuan Wang
  • Wenqiao Zang
  • Yuwen Du
  • Min Li
  • Guoqiang Zhao
Research Article

Abstract

RING box protein1 (RBX1), an essential component of SCF E3 ubiquitin ligases, plays an important role in gastric cancer. In the study, miR-194 and RBX1 expression was evaluated in 76 pairs of gastric tumor and non-tumor tissue samples by qRT-PCR, and clinicopathological characteristics were analyzed. CCK8, transwell assay, wound healing assay, and flow cytometry assay were performed to evaluate the effect of miR-194 on gastric cancer (GC) cellular proliferation, invasion, migration, apoptosis, and cell cycle, respectively. Luciferase reporter assays and Western blotting were used to evaluate whether RBX1 is a direct target of miR-194. The Kaplan-Meier method and log-rank test were used to evaluate the correlation between miR-194 or RBX1 expression and patient survival. Then, we found that miR-194 was significantly downregulated and RBX1 upregulated in GC tissues; both of which showed significant association with tumor size, location, invasion, and tumor node metastasis. Cell proliferation, invasion, and migration were significantly restricted with miR-194 overexpression. miR-194 downregulated RBX1 protein expression, and luciferase assays showed that binding sites in the RBX1 3′UTR were required for miR-194-mediated repression of RBX1, indicating that RBX1 was a direct target of miR-194. Transfection of RBX1 without the 3′UTR restored the miR-194-inhibiting migration function. miR-194 overexpression or RBX1 lowexpression was associated with prolonged survival of GC patients. In conclusion, upregulation of miR-194 can inhibit proliferation, migration, and invasion of GC cells, possibly by targeting RBX1. Aberrant expression of miR-194 and RBX1 is correlated to GC patient survival time.

Keywords

miR-194 RBX1 Migration Proliferation Survival time Gastric cancer 

Notes

Acknowledgments

This study was supported by the National Natural Science Foundation of China (No. 81272188 and No. 81301726) and the Education Department of Henan province science and technology research key projects (14A310002).

Conflicts of interest

None

References

  1. 1.
    Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 2010;127:2893–917.CrossRefPubMedGoogle Scholar
  2. 2.
    Jemal A, Siegel R, Ward E, Murray T, Xu J, Thun MJ. Cancer statistics, 2007. CA Cancer J Clin. 2007;57:43–66.CrossRefPubMedGoogle Scholar
  3. 3.
    Crew KD, Neugut AI. Epidemiology of gastric cancer. World J Gastroenterol. 2006;12:354–62.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Iravani O, Tay BW, Chua PJ, Yip GW, Bay BH. Claudins and gastric carcinogenesis. Exp Biol Med. 2013;238:344–9.CrossRefGoogle Scholar
  5. 5.
    Jang BG, Kim WH. Molecular pathology of gastric carcinoma. Pathobiology. 2011;78:302–10.CrossRefPubMedGoogle Scholar
  6. 6.
    Maurel M, Jalvy S, Ladeiro Y, Combe C, Vachet L, Sagliocco F, et al. A functional screening identifies five microRNAs controlling glypican-3: role of miR-1271 down-regulation in hepatocellular carcinoma. Hepatology. 2013;57:195–204.CrossRefPubMedGoogle Scholar
  7. 7.
    Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell. 1993;75:843–54.CrossRefPubMedGoogle Scholar
  8. 8.
    Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116:281–97.CrossRefPubMedGoogle Scholar
  9. 9.
    Garzon R, Calin GA, Croce CM. MicroRNAs in cancer. Annu Rev Med. 2009;60:167–79.CrossRefPubMedGoogle Scholar
  10. 10.
    Zhang S-J, Feng J-F, Wang L, Guo W, Du Y-W, Ming L, et al. miR-1303 targets claudin-18 gene to modulate proliferation and invasion of gastric cancer cells. Dig Dis Sci. 2014;59:1754–63.CrossRefPubMedGoogle Scholar
  11. 11.
    Yao Y, Suo AL, Li ZF, Liu LY, Tian T, Ni L, et al. MicroRNA profiling of human gastric cancer. Mol Med Rep. 2009;2:963–70.PubMedGoogle Scholar
  12. 12.
    Calin GA, Croce CM. MicroRNA signatures in human cancers. Nat Rev Cancer. 2006;6:857–66.CrossRefPubMedGoogle Scholar
  13. 13.
    Kerscher O, Felberbaum R, Hochstrasser M. Modification of proteins by ubiquitin and ubiquitin-like proteins. Annu Rev Cell Dev Biol. 2006;22:159–80.CrossRefPubMedGoogle Scholar
  14. 14.
    Skowyra D, Koepp DM, Kamura T, Conrad MN, Conaway RC, Conaway JW, et al. Reconstitution of G1 cyclin ubiquitination with complexes containing SCFGrr1 and Rbx1. Science. 1999;284:662–5.CrossRefPubMedGoogle Scholar
  15. 15.
    Frescas D, Pagano M. Deregulated proteolysis by the F-box proteins SKP2 and beta-TrCP: tipping the scales of cancer. Nat Rev Cancer. 2008;8:438–49.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Masuda TA, Inoue H, Sonoda H, Mine S, Yoshikawa Y, Nakayama K, et al. Clinical and biological significance of S-phase kinase-associated protein 2 (Skp2) gene expression in gastric carcinoma: modulation of malignant phenotype by Skp2 overexpression, possibly via p27 proteolysis. Cancer Res. 2002;62:3819–25.PubMedGoogle Scholar
  17. 17.
    Welcker M, Clurman BE. FBW7 ubiquitin ligase: a tumour suppressor at the crossroads of cell division, growth and differentiation. Nat Rev Cancer. 2008;8:83–93.CrossRefPubMedGoogle Scholar
  18. 18.
    Bai J, Zhou Y, Chen G, Zeng J, Ding J, Tan Y, et al. Overexpression of Cullin1 is associated with poor prognosis of patients with gastric cancer. Hum Pathol. 2011;42:375–83.CrossRefPubMedGoogle Scholar
  19. 19.
    Birner P, Schoppmann A, Schindl M, Dinhof C, Jesch B, Berghoff AS, et al. Human homologue for Caenorhabditis elegans CUL-4 protein overexpression is associated with malignant potential of epithelial ovarian tumours and poor outcome in carcinoma. J Clin Pathol. 2012;65:507–11.CrossRefPubMedGoogle Scholar
  20. 20.
    Song Y, Zhao F, Wang Z, Liu Z, Chiang Y, Xu Y, et al. Inverse association between miR-194 expression and tumor invasion in gastric cancer. Ann Surg Oncol. 2012;19:S509–17.CrossRefPubMedGoogle Scholar
  21. 21.
    Migita K, Takayama T, Matsumoto S, Wakatsuki K, Tanaka T, Ito M, et al. Prognostic impact of RING box protein-1 (RBX1) expression in gastric cancer. Gastric Cancer. 2014;17:601–9.CrossRefPubMedGoogle Scholar
  22. 22.
    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative pcr and the 2 (delta delta c (T)) method. Methods. 2001;25:402–8.CrossRefPubMedGoogle Scholar
  23. 23.
    Wang CJ, Zhou ZG, Wang L, Yang L, Zhou B, Gu J, et al. Clinicopathological significance of microRNA-31, -143 and -145 expression in colorectal cancer. Dis Markers. 2009;26:27–34.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Yan LX, Huang XF, Shao Q, Huang MY, Deng L, Wu QL, et al. MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis. RNA. 2008;14:2348–60.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Petroski MD, Deshaies RJ. Function and regulation of cullin-RING ubiquitin ligases. Nat Rev Mol Cell Biol. 2005;6:9–20.CrossRefPubMedGoogle Scholar
  26. 26.
    Yi Sun MT, Duan H, Swaroop M. SAG/ROC/Rbx/Hrt, a zinc RING finger gene family: molecular cloning, biochemical properties, and biological functions. Antioxid Redox Signal. 2001;3:635–50.CrossRefGoogle Scholar
  27. 27.
    Tan M, Davis SW, Saunders TL, Zhu Y, Sun Y. RBX1/ROC1 disruption results in early embryonic lethality due to proliferation failure, partially rescued by simultaneous loss of p27. Proc Natl Acad Sci. 2009;106:6203–8.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Wei D, Sun Y. Small RING finger proteins RBX1 and RBX2 of SCF E3 ubiquitin ligases: the role in cancer and as cancer targets. Gene Cancer. 2010;1:700–7.CrossRefGoogle Scholar
  29. 29.
    Jia L, Sun Y. SCF E3 ubiquitin ligases as anticancer targets. Curr Cancer Drug Targets. 2011;11:347–56.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Hino K, Tsuchiya K, Fukao T, Kiga K, Okamoto R, Kanai T, et al. Inducible expression of microRNA-194 is regulated by HNF-1alpha during intestinal epithelial cell differentiation. RNA. 2008;14:1433–42.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Meng Z, Fu X, Chen X, Zeng S, Tian Y, Jove R, et al. MiR-194 is a marker of hepatic epithelial cells and suppresses metastasis of liver cancer cells in mice. Hepatology. 2010;52:2148–57.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Xiaonan Chen
    • 1
  • Yuanyuan Wang
    • 1
  • Wenqiao Zang
    • 1
  • Yuwen Du
    • 1
  • Min Li
    • 1
  • Guoqiang Zhao
    • 1
  1. 1.College of Basic Medical SciencesZhengzhou UniversityZhengzhouChina

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