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Objective: To investigate shRNA interference PARG mouse colorectal cancer CT26 cells after expression of platelet adhesion of cancer cells and its mechanism. METHODS: The fragment carrying PARG-shRNA lentivirus interference (lentivirus) vector transfected murine colon carcinoma CT26 strains of PARG implementation silence interference. Have not been treated with CT26 cells were used as negative control group (blank control group), transfected with the empty vector lentiviral vector CT26 cells as a control group (transfected with empty viral particles control group), transfected with PARG-shRNAi lentivirus CT26 cells as PARG silent group (PARG-shRNAi group), Real time PCR assay after transfection PARG-shRNA CT26 cells PARG mRNA expression to determine the silencing effect. Staining using living cells, the morphological observation CT26 cells and platelet adhesion. CT26 cells by Western blot in PARG, PARP, Pi-akt, NF-κB, P-selectin and ICAM-1 expression was confirmed PARG by Pi-akt signaling pathway regulates downstream effectors, with the specific inhibition of phosphorylation of akt PARG silencing agent LY294002 treatment group CT26 cells, as inhibition of phosphorylation of akt group (Pi-akt restrain group). Results: In real time PCR and western blot experiments, compared with the control group, PARG PARG CT26 cells expressing silent group were significantly lower (P lt; 0.05), while the negative control group PARG expression with the empty vector control group ratio did not change significantly (P gt; 0.05) 2. in adhesion experiments, with the negative control group compared to the vector control CT26 cells and platelet adhesion was no significant change (P gt; 0.05). Compared with negative control group, PARG silencing group CT26 cells and platelet adhesion was significantly lower (P lt; 0.05). Compared with PARG silence, phosphorylated akt inhibited group CT26 cells and platelet adhesion rate increased significantly (P lt; 0.05). 3 In the WB experiment, with the negative control group compared to the vector control CT26 cells PARG, PARP, Pi-akt, NF-κB, ICAM-1, P-selectin expression did not change significantly (P gt; 0.05) . Compared with the negative control group, PARG silencing group CT26 cells PARG, PARP, NF-κB, ICAM-1, P-selectin was significantly lower (P lt; 0.05), Pi-akt significantly higher (P lt; 0.05). PARG silencing cells and compared akt phosphorylation inhibitor treated CT26 cells Pi-akt protein expression was significantly decreased (P lt; 0.05), and NF-κB, ICAM-1, P-selectin was significantly increased (P lt; 0.05). Conclusions: 1. PARG-shRNA Lentiviral transfection of mouse CT26 colorectal cancer cells, CT26 cells successfully silenced the expression of PARG, wait a minute viral vector itself within the protein expression of CT26 cells, biological behavior is not affected. 2, inhibition of PARG can significantly inhibit colorectal cancer CT26 cells and platelet adhesion. Tip: PARG thrombus formation may play an important role. 3. PARG inhibition of CT26 cells, the PARG, PARP, NF-κB, ICAM-1, P-selectin decreased, Pi-akt increased expression, suggesting: PARG inhibition of CT26 cells and platelets reduces the adhesion between, probably by downward PARP, increased Pi-akt signaling pathway, inhibition of NF-κB, thereby reducing ICAM-1, P-selectin expression and implementation.
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