Ji-Su Mo1,2, Seol-Hee Han1, Ki-Jung Yun1, Soo-Cheon Chae3,4. 1. Department of Pathology, School of Medicine, Wonkwang University, Iksan, Chonbuk, 54538, Republic of Korea. 2. Digestive Disease Research Institute, Wonkwang University, Iksan, Chonbuk, 54538, Republic of Korea. 3. Department of Pathology, School of Medicine, Wonkwang University, Iksan, Chonbuk, 54538, Republic of Korea. chaesc@wku.ac.kr. 4. Digestive Disease Research Institute, Wonkwang University, Iksan, Chonbuk, 54538, Republic of Korea. chaesc@wku.ac.kr.
Abstract
OBJECTIVE AND DESIGN: MicroRNAs (miRNAs) play an important role in the pathogenesis of human diseases by regulating the expression of target genes in specific cells or tissues. In this study, we analyzed the association between the MIR429 and its target gene, charged multivesicular body protein 5 (CHMP5), in human colon cancer cells and in a DSS-induced colitis mouse model. MATERIALS AND METHODS: A luciferase reporter system was used to confirm the effect of MIR429 on CHMP5 expression. Protein or mRNA expression of the target gene and associated molecules were measured by Western blot or quantitative RT-PCR (qRT-PCR), respectively. Flow cytometry was used to compare cell viability or cell cycle progression. RESULTS: CHMP5 mRNA and protein expression was directly down-regulated by MIR429. We found that MIR429 inhibited colon cancer cell growth and cell cycle progression, and CHMP5 was overexpressed in the DSS-induced colitis mouse model and human ulcerative colitis (UC) tissues. CONCLUSIONS: Our findings show that CHMP5 is a direct target of MIR429 in human colon cancer cell lines and suggest that CHMP5 up-regulation as a result of reduced MIR429 expression in DSS-induced mice colitis tissues and human UC tissues may restrict apoptosis and promote cell proliferation.
OBJECTIVE AND DESIGN: MicroRNAs (miRNAs) play an important role in the pathogenesis of human diseases by regulating the expression of target genes in specific cells or tissues. In this study, we analyzed the association between the MIR429 and its target gene, charged multivesicular body protein 5 (CHMP5), in humancolon cancer cells and in a DSS-induced colitismouse model. MATERIALS AND METHODS: A luciferase reporter system was used to confirm the effect of MIR429 on CHMP5 expression. Protein or mRNA expression of the target gene and associated molecules were measured by Western blot or quantitative RT-PCR (qRT-PCR), respectively. Flow cytometry was used to compare cell viability or cell cycle progression. RESULTS:CHMP5 mRNA and protein expression was directly down-regulated by MIR429. We found that MIR429 inhibited colon cancer cell growth and cell cycle progression, and CHMP5 was overexpressed in the DSS-induced colitismouse model and humanulcerative colitis (UC) tissues. CONCLUSIONS: Our findings show that CHMP5 is a direct target of MIR429 in humancolon cancer cell lines and suggest that CHMP5 up-regulation as a result of reduced MIR429 expression in DSS-induced micecolitis tissues and human UC tissues may restrict apoptosis and promote cell proliferation.