miR-125靶向SMAD4抑制脂多糖诱导的人结肠上皮细胞向肿瘤细胞转化

目的 观察脂多糖(LPS)对正常结肠上皮细胞向肿瘤细胞转化的影响,探索miR-125和SMAD4在细胞转化过程中的作用和机制.方法 以人正常结肠上皮细胞系HCoEpiCs为研究对象,用CCK-8法探索LPS的最佳刺激浓度;用LPS连续刺激HCoEpiCs细胞40代并检测细胞周期相关基因cyclinD1、癌基因c-myc...

miR-125 attenuates LPS-induced transition of colorectal epithelial cells into cancer cells through targeting SMAD4

Objective To investigate potential roles of miR-125 and SMAD4 in lipopolysaccharide(LPS)-induced colorectal epithelial cells transformed to cancer cells.Methods Cell viability of HCoEpiCs cells exposed to LPS for 24 h was evaluated by CCK-8 assay.CyclinD1,c-myc,Bcl-2,miR-125,and SMAD4 were measured by q-PCR in LPS-induced colorectal epithelial cells malignant transformation.Plane cloning and soft agar colony formation assay were used to identify the result of malignant transformation.Synthetic miR-125 mimic and inhibitor were used to mediate the expression of miR-125 in the 40th passage of LPS-induced HCoEpiCs cells.The activity of miR-125 in the region of SMAD43′-UTR was detected by the double luciferase gene report.Results Cell viability was signifi-cantly repressed by 10μg/mL LPS.The expression of miR-125 was increased in the 40th passage of LPS-induced HCoEpiCs cells,but the level of SMAD4 was down-regulated.The level of SMAD4 and c-myc were decreased when miR-125 was over-expressed in HCoEpiCs cells.Meanwhile,compared to the control group,the activity of SMAD43′-UTR-wild was repressed by miR-125 mimic,but no significant change was observed in the 3′-UTR-mut cells.Conclusions LPS-induced colorectal epithelial cells might be inhibited to transform to cancer cells through miR-125 binding to SMAD4.