Tollip Promotes The Growth And Metastasis Of Colon Cancer

Objective:To investigate the regulatory role and significance of Tollip in colon cancer.Methods:30 specimens of colon cancer and adjacent tissues were collected.The expressions of Tollip and related factors TGF-?1,Smad,E-cadherin and Vimentin were detected by immunohistochemistry(IHC),real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blotting respectively.the target RNA(Tollip-RNAi and Tollip-RNA)was transfected into colon cancer HT29 cells through lentiviral vectors to construct differentially expressed Tollip cells.KD,OE,NC and CON groups were set up.MTT,CCK-8 and clone formation experiments were used to detect the cell proliferation rate of each group under the effect of Tollip gene.Scratch test was used to determine the cell migration ability of each group.HT29 colon cancer cells differentially expressed in Tollip were injected subcutaneously into nude mice to construct tumor model,and the growth curve of tumors was drawn.At the 4th week,all nude mice were sacrificed and the average mass and volume of tumors in each group were compared,and HE staining was performed.Results:Tollip was diffusely expressed in the cell membrane and cytoplasm of colon cancer and adjacent intestinal mucosa,but not in the nucleus.The results of immunoblotting showed that the expression of Tollip in colon cancer group was significantly higher than that in adjacent group(P<0.05);RT-qPCR showed that the expression of Tollip in colon cancer group was significantly higher than that in adjacent group(P<0.05);Tollip was positively correlated with the expression of TGF-?1,Smad2 and Smad3(r=0.692,P<0.0001;r=0.480,P=0.032;r=0.520,P=0.006);There was no correlation with the expression of Vimentin and E-cadherin(P>0.05).There was no significant difference in the expression of Tollip in TNM stage(P>0.05).MTT,CCK-8 method and clone formation test after RNA transfection showed that KD group was lower than CON group in cell proliferation characteristics(P<0.05),KD group and NC group lacked statistical difference(P>0.05);OE group was higher than CON group and NC group(P<0.05).Scratch experiment showed that the mobility of cells in KD group was significantly lower than that in CON group(P<0.05).There was no statistical difference between KD group and NC group(P>0.05).The mobility of cells in OE group was significantly higher than that in CON group and NC group(P<0.05).tumor volume was measured by tumorigenesis in nude mice,but there was no statistical difference between KD group and CON group and NC group(P>0.05).Compared with CON group and NC group,the volume of OE group increased significantly in 3 and 4 weeks(P<0.05).The weight of tumors in KD group was significantly lower than that in CON group and NC group(P<0.05),while that in OE group was significantly higher than that in CON group and NC group(P<0.05).Conclusion:Tollip promotes the occurrence and development of colon tumors.Tollip is correlation with TGF-?1/Smad signaling pathway.Tollip promotes the regulation of colon tumors.