OBJECTIVETo detect miR-200a expression in human colorectal carcinnoma (CRC) cell lines and explore the role of miR-200a in regulating the biological behavior of CRC cells.

METHODSReal-time quantitative RT-PCR (qRT-PCR) was used to detect miR-200a expression levels in 6 CRC cell lines (HCT116, HT29, LS174T, SW480, SW620 and LoVo). miR-200a mimics were transiently transfected into LoVo, and the changes in cell proliferation, apoptosis, migration, and cell-cell adhesion were assessed using CCK-8 assay, TUNEL assay, transwell migration assay, and homogenous adhesion experiment, respectively.

RESULTSThe expression of miR-200a was down-regulated in the 6 CRC cell lines, among which the highly metastatic LoVo cell line showed the lowest expression and the tumorigenic but non-metastatic CRC cell line HCT116 had the highest expression. Overexpression of miR-200a depressed cell proliferation and migration but promoted cell apoptosis and cell-cell adhesion in LoVo cells.

CONCLUSIONmiR-200a plays a role in regulating the invasiveness and metastasis of CRC, and overexpression of miR-200a causes a significant reduction of cell proliferation and migration and promotes apoptosis and cell-cell adhesion in LoVo cells.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; metabolism ; Transfection

Objective To study the effect of pirfenidone (PFD) on the transformation of rat corneal stromal cells into fibroblasts in vitro and further explore the anti-fibrotic effect of PFD. Methods The corneal stromal cells from SD rat was isolated and cultured ,and was determined by vimentin stain. The experiment was divided into control group(DMEM+10%FBS),TGF-β1 group(2 ng/mL TGF-β1+DMEM+10%FBS)and PFD group(1 mg/mL PFD+ 2 ng/mL TGF-β1+DMEM+ 10%FBS). Cell proliferation was detected by CCK-8 assay. Collagen Ⅰ,Collagen Ⅲ,Keratocan and CD99 expression were detected by Western blot. Results Compared with control group and TGF-β1 group,the cell proliferation were significantly decreased in PFD group(P<0.05). Western blot showed that PFD can up-regulated Collagen Ⅰand Keratocan but down-regulated Collagen ⅢCD90 expression(P < 0.05). The ratio of Collagen Ⅲ/Collagen Ⅰ in PFD group was lowest in all groups(P < 0.05). Conclusion PFD can resistant fibration in corneal stromal cells may through the inhibition of TGF-β ,which affect the collagen synthesis and Keratocan,CD90 expression.