Sung Noh Hong1, Sung Jin Kim2, Eun-Ran Kim1, Dong Kyung Chang1, Young-Ho Kim1. 1. Department of Medicine, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea. 2. Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Republic of Korea.
Abstract
BACKGROUND: Genome-wide methylation arrays have revealed aberrant methylation of N-Myc downstream-regulated gene 2 (NDRG2) promoter in colorectal cancer (CRC). This study investigated the role of NDRG2 in colorectal carcinogenesis. METHODS: The aberrant promoter methylation, mRNA, and protein expression of NDRG2 were evaluated in 27 pairs of human CRC and adjacent normal tissues and seven human CRC-derived cell-lines. After stable NDRG2 over-expressed RKO and DLD-1 human CRC cell-lines were constructed, in vitro functional assays, including colony formation, cell viability, proliferation, invasion and migration assays, and in vivo xenograft models were performed. RESULTS: The promoter of NDRG2 was methylated in 89% human CRC tissue compared to adjacent normal colonic mucosa (7.4%; P < 0.001). High-level methylation of NDRG2 promoter was more prevalent in proximal CRC (P = 0.022) and advanced T stage (P = 0.039). NDRG2 mRNA and protein expression was down-regulated in 89% and 100% human CRC tissue, respectively. In human CRC cell-lines, the promoter of NDRG2 was methylated aberrantly and mRNA, and protein expression of NDRG2 was down-regulated. NDRG2 mRNA expression was reactivated by 5-aza-2'-deoxycytidine. Colony formation of NDRG2 over-expressing RKO cells was inhibited (P = 0.012), as was the viability, proliferation, and invasion of NDRG2 over-expressing DLD-1 cells (P < 0.001, P = 0.003, and P = 0.044, respectively). Tumor volume in xenograft mice transplanted with NDRG2 over-expressing RKO and DLD-1 cells was smaller than that in controls (P = 0.002 and P = 0.001, respectively). CONCLUSIONS: Epigenetic silencing of NDRG2 induces proliferation and invasion of CRC and may be associated with proximal CRC and advanced T stage. NDRG2 methylation might serve as novel biomarker of CRC.
BACKGROUND: Genome-wide methylation arrays have revealed aberrant methylation of N-Myc downstream-regulated gene 2 (NDRG2) promoter in colorectal cancer (CRC). This study investigated the role of NDRG2 in colorectal carcinogenesis. METHODS: The aberrant promoter methylation, mRNA, and protein expression of NDRG2 were evaluated in 27 pairs of human CRC and adjacent normal tissues and seven human CRC-derived cell-lines. After stable NDRG2 over-expressed RKO and DLD-1 human CRC cell-lines were constructed, in vitro functional assays, including colony formation, cell viability, proliferation, invasion and migration assays, and in vivo xenograft models were performed. RESULTS: The promoter of NDRG2 was methylated in 89% human CRC tissue compared to adjacent normal colonic mucosa (7.4%; P < 0.001). High-level methylation of NDRG2 promoter was more prevalent in proximal CRC (P = 0.022) and advanced T stage (P = 0.039). NDRG2 mRNA and protein expression was down-regulated in 89% and 100% human CRC tissue, respectively. In human CRC cell-lines, the promoter of NDRG2 was methylated aberrantly and mRNA, and protein expression of NDRG2 was down-regulated. NDRG2 mRNA expression was reactivated by 5-aza-2'-deoxycytidine. Colony formation of NDRG2 over-expressing RKO cells was inhibited (P = 0.012), as was the viability, proliferation, and invasion of NDRG2 over-expressing DLD-1 cells (P < 0.001, P = 0.003, and P = 0.044, respectively). Tumor volume in xenograft mice transplanted with NDRG2 over-expressing RKO and DLD-1 cells was smaller than that in controls (P = 0.002 and P = 0.001, respectively). CONCLUSIONS: Epigenetic silencing of NDRG2 induces proliferation and invasion of CRC and may be associated with proximal CRC and advanced T stage. NDRG2 methylation might serve as novel biomarker of CRC.
Authors: M Lango-Chavarría; G K Chimal-Ramírez; M E Ruiz-Tachiquín; N A Espinoza-Sánchez; M C Suárez-Arriaga; E M Fuentes-Pananá Journal: Int J Oncol Date: 2017-01-05 Impact factor: 5.650
Authors: Kyung Rok Kim; Kyung A Kim; Joon Sung Park; Jun Young Jang; Yuri Choi; Hyung Ho Lee; Dong Chul Lee; Kyung Chan Park; Young Il Yeom; Hyun-Jung Kim; Byung Woo Han Journal: Biomolecules Date: 2020-01-06