The Effect Of SHRNA Interferes PARG In Mouse Colon Carcinoma CT26 Cell On The Adhesion Of CT26 And Platelet

Objective: The aim is to discuss the effect of shRNA interferes PARG in mouse colon carcinoma CT26 cells on the adhesion of CT26 and platelets and it's possible mechanism.Methods: Lentivirus that carried PARG-shRNA section was used for transfecting CT26 cells, then CT26 cells with silenced PARG were perpetually selected. CT26 cells without any treatment served as blank control group;CT26 cells that treated with empty viral particles served as empty viral particles control group;CT26 cells transfected with PARG-shRNAi insert lentivirus served as PARG-shRNAi group. Real-time PCR was used for testing the effeciency of transfected experiment by semi-quantity method. Regarding the influence of silencing PARG in CT26 cells on the ability of adhesion of CT26 and platelets,viable CT26 cells and platelets were dyed for the test.Western blot assayed the expression of PARG,PARP,Pi-akt,NF-κB,P-selectin and ICAM-1,moreover, to further certify Pi-akt was the signal pathway in CT26 cells,the Pi-akt inhibitor LY294002 was used to treat the PARG-shRNA CT26 cells.Results:1.Both real time-PCR and Western Blotting results showed that the expression of PARG in the PARG-shRNAi group were obviously lower than that in the control group(P<0.05)and there was no distinct between blank control group and empty viral particles control group(P>0.05) .2. In the adhesion assay. The adhesion rate of platelets to CT26 cells in PARG-shRNAi groups was reduced, when compared with blank control group(P<0.05). The adhesion rate was not obvious changed when transfected with empty viral particles control group compared with blank control group(P>0.05). The adhesion rate was a statistically valid decrease in Pi-akt restrained group when compared with PARG-shRNAi group(P<0.05).3. The expressions of PARG,PARP,NF-κB,P-selectin and ICAM-1 were reduced in PARG-shRNAi group compared to blank control group. Simultaneously, Pi-akt was up-regulated (P<0.05); There were no evident change when transfected with empty viral particles control group compared with blank control group(P>0.05). The exression of Pi-akt in Pi-akt restrained group was reduced and the down-stream factors NF-κB,ICAM-1,P-selectin were up-regulated when compared to PARG-shRNAi group (P<0.05).Conclusion:1. The PARG-silenced CT26 cells could suppress the CT26 cells adhere to platelets,and lentvirus itself didn't influence CT26 cells on adhesion ability. It suggests that PARG may play an important role in the process of cancer embolus forming.2. Decrease of PARG,PARP, NF-κB,ICAM-1,P-selectin and rise of Pi-akt in PARG-silenced CT26 cells suggested that PARG suppression could decrease the adhesion rate of platelets to CT26 cells,it probably relates to the following process: down-regulate PARP,up-regulate Pi-akt,then restrain NF-κB and finally decrease ICAM-1,P-selectin.