Effect Of ART1on Stress Fiber Play A Role In CT26Cellular Adhesion And Migration Ability

Objective: To explore the effects of mono(ADP-ribosyl)transferase-1（ART1）genesilencingandoverexpressiononstress fiber and theabilityofmouse colon carcinoma CT26cellular matrix adhension, migration andinvasion and its mechanism.Methods: mouse colon carcinoma CT26cells were infected by therecombinant Lentivirus of ART1-shRNA and ART1-overexpression.TheUntreated group and the Negative control group(NC-shRNA group) wereused as control groups, the ART1-shRNA group and theART1-overexpression group were used as experimental groups,Theexpression of ART1mRNA was detected by RT-PCR.The microfilamentmorphological changes was detected by laser scanning confocalmicroscopy.The matrix adhesion, migration and invasion potencies ofvarious group CT26cells were observed by cell matrix adhesion,migration,invasion analysis. Liver metastasis models of BALB/c mice wereestablished by intrasplenic inoculation of Untreated,group CT26cells,ART1-shRNA group cells, ART1-overexpression group cells and NC-shRNA group cells respectively.The changes of splenic transplantationtumors, liver metastasis and survival time were observed。The expressions ofART1, RhoA, integrin β1, FAK in the CT26cells and splenic transplantationtumors were measured by Western blot analysis.Results:1The CT26cells with ART1gene silencing, ART1-overexpressions, Negative control-shRNA were obtained. RT-PCR andWestern blotting showed: the expressions of ART1mRNA and ART1inART1-shRNA group were significantly lower than Untreated group andNC-shRNA group(p<0.05), the expressions of ART1-overexpression groupwere significantly higher than Untreated group and NC-shRNAgroup(p<0.05), the difference between the Untreated group and NC-shRNAgroup was unsignificantly（P>0.05）.2The obviously difference of cell stress fiber formation was displayedby laser scanning confocal microscopy,The stress fiber formation ofART1-shRNA group were significantly lower than Untreated group andNC-shRNA group(p<0.05), the stress fiber formation of ART1-overexpression group were significantly higher than Untreated group andNC-shRNA group(p<0.05), the difference between the Untreated group andNC-shRNA group was unsignificantly（P>0.05）.The results of cell adhesion,migration and invasion assaies showed that the adhesion, migration andinvasion abilities of ART1-shRNA group were reduced, the adhesion,migration and invasion abilities of ART1-overexpression group were increased, when compared with Untreated group(P<0.05),the differencebetween the Untreated group and NC-shRNA group was unsignificantly（P>0.05）.3Succeeded to construct experimental metastasis model.The splenictransplantation tumors were observed in four groups,the average weight,volume of splenic transplantation tumors and the quantity of liver metastasisnodules of ART1-shRNA group were reduced, ART1-overexpression groupwere increased, when compared with Untreated group(P<0.05). and nosignificant difference between the Untreated group and NC-shRNA group (P>0.05).The Kaplan-Meier curves showed that the survival time ofART1-shRNA group mice was significantly prolonged (P<0.05), thesurvival time of ART1-overexpression group mice was significantly shorter(P<0.05), and no significant difference between the Untreated group andNC-shRNA group (P>0.05).4Western blotting results showed that the expressions of ART1, RhoA,integrin β1, FAK in the ART1-shRNA group CT26cells and splenictransplantation tumors were significantly lower than Untreatedgroup(P<0.05), The expressions of ART1-overexpression group weresignificantly higher than Untreated group (P<0.05), and no significantdifference between the Untreated group and NC-shRNA group (P>0.05).Conclusion: the experimental results showed that the ART1genesilencing can reduce the formation of stress fiber and then reduce the adhesion, movement abilities of CT26cells and inhibit the formation ofmouse spleen transplantation tumor and liver metastatic tumor nodules.ART1gene overexpression can promote the formation of stress fiber and theadhesion, movement abilities of CT26cells and promote the formation ofmouse spleen transplantation tumor and liver metastatic tumornodules.Suggesting that ART1may play a regulatory role in tumor cellinvasion process, which may be related to the expression of RhoA, integrin β1and FAK and the morphology and function of the microfilament...