Down-regulation of EZH2 by RNA Interference Inhibits Proliferation and Invasion of ACHN Cells via the Wnt/β-catenin Pathway

  • Yuan, Jun-Bin (Department of Urology, Xiangya Hospital, Central South University) ;
  • Yang, Luo-Yan (Department of Urology, The Second Xiangya Hospital, Central South University) ;
  • Tang, Zheng-Yan (Department of Urology, Xiangya Hospital, Central South University) ;
  • Zu, Xiong-Bing (Department of Urology, Xiangya Hospital, Central South University) ;
  • Qi, Lin (Department of Urology, Xiangya Hospital, Central South University)
  • Published : 2012.12.31


Although enhancer of zeste homolog 2 (EZH2) has been reported as an independent prognostic factor in renal cell carcinoma (RCC), little is known about the exact mechanism of EZH2 in promoting the genesis of RCC. However, several studies have shown that dysregulation of the Wnt/${\beta}$-catenin signaling pathway plays a crucial role. Therefore, we determined whether EZH2 could affect ACHN human RCC cell proliferation and invasion via the Wnt/${\beta}$-catenin pathway. In the present study, we investigated the effects of short interfering RNA (siRNA)-mediated EZH2 gene silencing on Wnt/${\beta}$-catenin signaling in ACHN cells. EZH2-siRNA markedly inhibited the proliferation and invasion capabilities of ACHN, while also reducing the expression of EZH2, Wnt3a and ${\beta}$-catenin. In contrast, cellular expression of GSK-$3{\beta}$ (glycogen synthase kinase-$3{\beta}$), an inhibitor of the Wnt/${\beta}$-catenin pathway, was conspicuously higher after transfection of EZH2 siRNA. These preliminary findings suggest EZH2 may promote proliferation and invasion of ACHN cells via action on the Wnt/${\beta}$-catenin signaling pathway.


EZH2;invasion;proliferation;renal cell carcinoma;RNA interference;Wnt/${\beta}$-catenin signaling


  1. Wagener N, Macher-Goeppinger S, Pritsch M, et al (2010). Enhancer of zeste homolog 2 (EZH2) expression is an independent prognostic factor in renal cell carcinoma. BMC Cancer, 10, 524-33.
  2. Wang JS, Ji AF, Wan HJ, et al (2012). Gene silencing of $\beta$-catenin by RNAi inhibits proliferation of human esophageal cancer cells by inducing G0/G1 cell cycle arrest. Asian Pac J Cancer Prev, 13, 2527-32.
  3. Wan X, Liu J, Lu JF, et al (2012). Activation of $\beta$-catenin signaling in androgen receptor-negative prostate cancer cells. Clin Cancer Res, 18, 726-36.
  4. Ciarapica R, Miele L, Giordano A, Locatelli F, Rota R (2011). Enhancer of zeste homolog 2 (EZH2) in pediatric soft tissue sarcomas: first implications. BMC Med, 9, 63-71.
  5. Clevers H (2006). Wnt/beta-catenin signaling in development and disease. Cell, 127, 469-80.
  6. Herold S, Herkert B, Eilers M (2009). Facilitating replication under stress: an oncogenic function of MYC? Nat Rev Cancer, 9, 441-4.
  7. Kleer CG, Cao Q, Varambally S, et al (2003). EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells. Proc Natl Acad Sci USA, 100, 11606-11.
  8. Li X, Gonzalez ME, Toy K, et al (2009). Targeted overexpression of EZH2 in the mammary gland disrupts ductal morphogenesis and causes epithelial hyperplasia. Am J Pathol, 175, 1246-54.
  9. Li Y, Pawlik B, Elcioglu N, et al (2010). LRP4 mutations alter Wnt/beta-catenin signaling and cause limb and kidney malformations in Cenani-Lenz syndrome. Am J Hum Genet, 86, 696-706.
  10. Liu C, Li Y, Semenov M, et al (2002). Control of beta-catenin phosphorylation/degradation by a dual-kinase mechanism. Cell, 108, 837-47.
  11. Miyata Y, Iwata T, Ohba K, et al (2006). Expression of matrix metalloproteinase-7 on cancer cells and tissue endothelial cells in renal cell carcinoma: prognostic implications and clinical significance for invasion and metastasis. Clin Cancer Res, 12, 6998-7003.
  12. Montgomery ND, Yee D, Montgomery SA, Magnuson T (2007). Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation. J Mol Biol, 374, 1145-57.
  13. Sher F, Rossler R, Brouwer N, et al (2008). Differentiation of neural stem cells into oligodendrocytes: involvement of the polycomb group protein Ezh2. Stem Cells, 26, 2875-83.
  14. Shi B, Liang J, Yang X, et al (2007). Integration of estrogen and Wnt signaling circuits by the polycomb group protein EZH2 in breast cancer cells. Mol Cell Biol, 27, 5105-19.
  15. Smalley MJ, Dale TC (1999). Wnt signalling in mammalian development and cancer. Cancer Metastasis Rev, 18, 215-30.
  16. Stocker G, Ott K, Henningsen N, et al (2009). CyclinD1 and interleukin-1 receptor antagonist polymorphisms are associated with prognosis in neoadjuvant-treated gastric carcinoma. Eur J Cancer, 45, 3326-35.
  17. Sudo T, Utsunomiya T, Mimori K, et al (2005). Clinicopathological significance of EZH2 mRNA expression in patients with hepatocellular carcinoma. Br J Cancer, 92, 1754-8.
  18. Thiel U, Pirson S, Müller-Spahn C, et al (2011). Specific recognition and inhibition of Ewing tumour growth by antigen-specific allo-restricted cytotoxic T cells. Br J Cancer, 104, 948-56.
  19. Wagener N, Holland D, Bulkescher J, et al (2008). The enhancer of zeste homolog 2 gene contributes to cell proliferation and apoptosis resistance in renal cell carcinoma cells. Int J Cancer, 123, 1545-50.
  20. Andersson ER, Bryjova L, Biris K, et al (2010). Genetic interaction between Lrp6 and Wnt5a during mouse development. Dev Dyn, 239, 237-45.
  21. Banumathy G, Cairns P (2010). Signaling pathways in renal cell carcinoma. Cancer Biol Ther, 10, 658-64.
  22. Bracken AP, Pasini D, Capra M, et al (2003). EZH2 is downstream of the pRB-E2F pathway, essential for proliferation and amplified in cancer. EMBO J, 22, 5323-35.
  23. Bryant RJ, Cross NA, Eaton CL, Hamdy FC, Cunliffe VT (2007). EZH2 promotes proliferation and invasiveness of prostate cancer cells. Prostate, 67, 547-56.
  24. Cardoso C, Mignon C, Hetet G, et al (2000). The human EZH2 gene: genomic organisation and revised mapping in 7q35 within the critical region for malignant myeloid disorders. Eur J Hum Genet, 8, 174-80.
  25. Chen AE, Ginty DD, Fan CM (2005). Protein kinase A signalling via CREB controls myogenesis induced by Wnt proteins. Nature, 433, 317-22.
  26. Cheng AS, Lau SS, Chen Y, et al (2011). EZH2-mediated concordant repression of Wnt antagonists promotes $\beta$-catenin-dependent hepatocarcinogenesis. Cancer Res, 71, 4028-39.
  27. Choi H, Chun YS, Kim TY, Park JW (2010). HIF-2alpha enhances beta-catenin/TCF-driven transcription by interacting with beta-catenin. Cancer Res, 70, 10101-11.

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