The Silencing of CCND2 by Promoter Aberrant Methylation in Renal Cell Cancer and Analysis of the Correlation between CCND2 Methylation Status and Clinical Features

PLoS One. 2016 Sep 1;11(9):e0161859. doi: 10.1371/journal.pone.0161859. eCollection 2016.

Abstract

Cyclin D2 (CCND2) is a member of the D-type cyclins, which plays a pivotal role in cell cycle regulation, differentiation and malignant transformation. However, its expression status and relative regulation mechanism remains unclear in renal cell cancer (RCC). In our study, the mRNA expression level of CCND2 is down-regulated in 22/23 paired RCC tissues (p<0.05). In addition, its protein expression level is also decreased in 43/43 RCC tumor tissues compared with its corresponding non-malignant tissues (p<0.001). We further detected that CCND2 was down-regulated or silenced in 6/7 RCC cell lines, but expressed in "normal" human proximal tubular (HK-2) cell line. Subsequently, MSP and BGS results showed that the methylation status in CCND2 promoter region is closely associated with its expression level in RCC cell lines. Treatment with 5-Aza with or without TSA restored CCND2 expression in several methylated RCC cell lines. Among the 102 RCC tumors, methylation of CCND2 was detected in 29/102 (28%) cases. Only 2/23 (8.7%) adjacent non-malignant tissues showed methylation. We then analyzed the correlation of clinical features and its promoter methylation. Collectively, our data suggested that loss of CCND2 expression is closely associated with the promoter aberrant methylation.

MeSH terms

  • Carcinoma, Renal Cell / genetics*
  • Carcinoma, Renal Cell / pathology*
  • Cell Line, Tumor
  • Cell Proliferation
  • Cyclin D2 / genetics*
  • DNA Methylation*
  • Gene Silencing*
  • Genome
  • Humans
  • Kidney Neoplasms / genetics*
  • Kidney Neoplasms / pathology*
  • Promoter Regions, Genetic*

Substances

  • CCND2 protein, human
  • Cyclin D2

Grants and funding

This study was supported by grants from the National Natural Science Foundation (No. 81272290, No. 81572510, No. 81370858 and No. 81570683), Beijing Municipal Science and Technology Commission (No. Z131107002213130) and Beijing Natural Science Foundation (No. 7142161).