ABSTRACT
Objective To investigate the effect of Nestin gene silencing on the proliferation of human esophageal cancer ECA109 cells and possible mechanism .Methods Lenti-Nestin was constructed and transfected into ECA109 cells to establish a stable Nestin-silencing cell line Lenti-Nestin.Blank group , scrambled group , Lenti-Nestin group were set up .The expressions of Nestin , c-myc and cyclin D1 mRNA and protein levels were detected by qRT-PCR and Western blot .The cell proliferation was analyzed by CCK 8 assay .Results The stable Nestin-si-lencing cell line was successfully established .The expression of Nestin mRNA ( P<0.01 ) and protein ( P<0.05 ) levels were reduced significantly and the downregulation evidently suppressed cell proliferation ( P<0.01 ) and col-ony forming capacity ( P<0.05 ) compare with the scrambled group and blank group .However ,The c-myc and cy-clin D1 expression levels in ECA 109 cells in Lenti-Nestin group were significantly lower than that of scrambled group and blank group ( P<0.05 ) .Conclusions Knockdown Nestin expression significantly inhibits the level of esophageal cancer ECA109 cells proliferation,which may act via influencing the expression of c-myc, cyclin D1.
ABSTRACT
AIM:ToinvestigatetheeffectofsmallinterferenceRNA(siRNA)-mediatedsilencingofnestin gene on the invasion and migration of human esophageal cancer ECA 109 cells and the possible mechanism .METHODS:The esophageal cell line ECA 109 was transfected with siRNA targeting nestin and the cell invasion and migration abilities were observed.The expression of nestin, MMP2, MMP9, VEGF, and total and nuclear β-catenin proteins in the transfec-ted cells were determined by real-time PCR and Western blot .RESULTS:Compared with control group , the expression of nestin at mRNA and protein levels was significantly down-regulated in the ECA109 cells transfected with nestin-siRNA, so was the expression of MMP2, MMP9, VEGF, and total and nuclear β-catenin proteins.The levels of invasion and migra-tion capacities of ECA 109 cells transfected with nestin-siRNA were lower than those in the cells transfected with control-siRNA.CONCLUSION:Knockdown of nestin expression significantly inhibits the invasion and migration of the esophage-al cancer cells , which may act via suppressing β-catenin translocation to the nucleus and influencing the expression of MMP2, MMP9 and VEGF.
ABSTRACT
[ ABSTRACT] AIM:To investigate the effect of DEC1 gene over-expression on the proliferation and invasion abili-ties of human esophageal cancer ECA109 cells.METHODS: ECA109 cells were transfected with plasmid pcDNA3.1 (-)/DEC1 (DEC1 group) or pcDNA3.1 (-) (vector group).The mRNA and protein levels of DEC1, cyclin D1 and MMP-9 were evaluated by real-time PCR and Western blot, respectively.The effects of DEC1 over-expression on the prolif-eration and invasion abilities of the ECA109 cells were evaluated by CCK-8 assay, colony formation assay and Transwell test respectively.RESULTS:The DEC1 expression level in ECA109 cells in DEC1 group was significantly higher than that in vector group (P<0.01), but the levels of MMP9 and cyclin D1 expression were opposite (P<0.01).However, both the proliferation and invasion abilities of ECA109 cells in DEC1 groups decreased significantly as compared with those in vector group (P<0.05).CONCLUSION:The over-expression of DEC1 significantly inhibits the proliferation and invasion of ECA109 cells, which may be involved in the expression of cyclin D1 and MMP9.