ABSTRACT
Objective To explore the effects and regulation mechanism of miR-489 on the stem cell phenotype and epithelial-mesenchymal transition of colon cancer cells in vitro.Methods The miR-489 expression of colon cancer cell lines ( HT29, SW480 and SW620, HCT116) and normal intestinal epithelial cells HIEC were detec-ted by RT-qPCR, the miR-489 expression in colon cancer cells was raised by gene transfection technology, the colon cancer stem cell markers CD133, CD44, EpCAM, ALDH1 and epithelial marker E-cadherin, mesenchy-mal cells markers vimentin and N-cadherin were detected by Western blot.The expression of TWIST1 mRNA and protein were detected by RT-qPCR and Western blot.Results The miR-489 relative expression in four kinds of colon cancer cells ( HT29, SW480 and SW620, HCT116) were significantly lower than that of normal HIEC in-testinal epithelial cells, the difference was statistically significant ( P<0.05) .Up-regulation of miR-489 expres-sion in HT29 and HCT116 cells leaded to lower expression of colon cancer stem cell marker CD133, CD44, EpCAM ALDH1 and mesenchymal cells markers Vimentin , N-cadherin, higher expression of epithelial markers ;E-cadherin (P<0.05).Also, up-regulation of miR-489 expression in HT29 and HCT116 cells leaded to lower ex-pression of TWIST 1 mRNA and protein ( P<0.05 ) .Conclusions miR-489 is down-regulated in colon cancer cells and up-regulation of miR-489 expression inhibits stem cell phenotype and epithelial-mesenchymal transition through targeting TWIST1 in colon cancer .
ABSTRACT
Objective@#To explore the potential therapeutic role of miR-489 in silica-induced pulmonary fibrosis mouse models.@*Methods@#A total of 32 C57BL/6 male mice were randomly divided into four groups: saline, silica, silica plus miRNA control and silica plus miR-489 agomir (n=8 in each group) . The mice were instilled with silica particles suspended in saline or sterile saline intratracheally. Subsequently, miR-489 agomir or miRNA control was injected via the tail vein into each mouse at days 28, 35, 42 and 49, the miR-489 levels, histological examination, collagen deposition, fibrotic biomarkers (E-cadherin, α-SMA, Vimentin, Fibronectin) and transforming growth factor-β1 (TGF-β1) protein levels in mouse lung tissues were measured.@*Results@#miR-489 levels in silica plus miR-489 group were significantly increased in lung tissues compared with silica plus miRNA control group (P<0.05) . Histological examination showed attenuated inflammation, less severe fibrotic foci and less destruction of alveolar architecture in the silica plus miR-489 group. Additionally, both the severity and distribution of lung lesions were ameliorated in silica plus miR-489 group compared with the silica plus miRNA control group (P<0.05) . The collagen deposition and hydroxyproline levels in silica plus miR-489 group were significantly decreased compared with the silica plus miRNA control group (P<0.05) . These changes were supported by decreased protein levels of α-SMA, Vimentin, Fibronectin, TGF-β1 along with increased protein levels of E-cadherin in silica plus miR-489 group (P<0.05) .@*Conclusion@#Our data indicate that the upregulation of miR-489 has potential therapeutic role in silica-induced pulmonary fibrosis in vivo, which may be associated with the depression of TGF-β1 release.