Department of Pathology, School of Medicine, Wonkwang University, Iksan, Chonbuk, 54538, Republic of Korea.
Digestive Disease Research Institute, Wonkwang University, Iksan, Chonbuk, 54538, Republic of Korea.
Inflamm Res. 2018 Dec;67(11-12):985-996. doi: 10.1007/s00011-018-1194-z. Epub 2018 Oct 17.
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.
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.
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.
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.
微小 RNA（miRNAs）通过在特定细胞或组织中调节靶基因的表达，在人类疾病的发病机制中发挥重要作用。在这项研究中，我们分析了 MIR429 及其靶基因电荷多泡体蛋白 5（CHMP5）在人结肠癌细胞和 DSS 诱导的结肠炎小鼠模型中的关联。
使用荧光素酶报告系统来确认 MIR429 对 CHMP5 表达的影响。通过 Western blot 或定量 RT-PCR（qRT-PCR）分别测量靶基因和相关分子的蛋白或 mRNA 表达。通过流式细胞术比较细胞活力或细胞周期进程。
CHMP5 mRNA 和蛋白表达被 MIR429 直接下调。我们发现 MIR429 抑制结肠癌细胞生长和细胞周期进程，并且在 DSS 诱导的结肠炎小鼠模型和人类溃疡性结肠炎（UC）组织中 CHMP5 过表达。
我们的研究结果表明，CHMP5 是人类结肠癌细胞系中 MIR429 的直接靶标，并表明由于 DSS 诱导的小鼠结肠炎组织和人类 UC 组织中 MIR429 表达减少，CHMP5 的上调可能限制细胞凋亡并促进细胞增殖。
