miR-93 suppresses proliferation and colony formation of human colon cancer stem cells.

摘要: AIM: To identify differentially expressed microRNAs (miRNAs) in human colon cancer stem cells (SW1116csc) and study their function SW1116csc proliferation. METHODS: were isolated from the cell line, SW1116 cultured serum-free medium. A miRNA microarray was used to detect differential expression profiles of miRNAs cells. Real-time quantitative polymerase chain reaction (PCR) performed verify candidate obtained microarray. Target mRNAs predicted with target prediction tools. plasmids transfected into using Lipofectamine 2000 reagent. Cell proliferation curves generated trypan blue staining, colony formation rate measured soft agar assay. Expression proteins detected reverse transcription (RT)-PCR western blotting. RESULTS: Compared cells, 35 (including hsa-miR-192, hsa-miR-29b, hsa-miR-215, hsa-miR-194, hsa-miR-33a hsa-miR-32) upregulated more than 1.5-fold, 11 hsa-miR-93, hsa-miR-1231, hsa-miRPlus-F1080, hsa-miR-524-3p, hsa-miR-886-3p hsa-miR-561) downregulated SW1116csc. The results further validated RT-PCR. miR-93 downregulated, its mRNA targets included BAMBI, CCND2, CDKN1A, HDAC8, KIF23, MAP3K9, MAP3K11, MYCN, PPARD, TLE4 ZDHHC1. Overexpressed significantly inhibited by Furthermore, negatively regulated protein levels HDAC8 TLE4. CONCLUSION: Some during differentiation may inhibit formation.