Expression Of PRR11 In Colorectal Cancer And Its Effect On Progression Of Colorectal Cancer

Objective:To detect the expression of proline-rich protein 11(PRR11)in colorectal cancer and adjacent tissues,colorectal cancer cells and normal colorectal epithelial cells,and to detect the impact of PRR11 overexpression and silencing on the invasion and metastasis of colorectal cancer cells,as well as the related signals of epithelial-mesenchymal transition(EMT).The relationship between the expression level and the clinicopathological factors of colorectal cancer and the relationship between the expression level and the invasion and metastasis of colorectal cancer were studied.Methods:The expression of PRR11,?-catenin,E-cadherin and vimentin in colorectal cancer and its adjacent tissues were detected by immunohistochemistry SP method.We use western blot and RT-PCR to detect the expression of PRR11 in three kinds of colorectal cancer(SW480,HT29,HCT116)and one kind of normal colorectal epithelial cell(HCoEpiC).After overexpression and silencing of PRR11 in colorectal cancer cell HT29,Western blot method was used to detect the effect of overexpression and silencing of PRR11 on the expression of PRR11,?-catenin,E-cadherin and vimentin.RT-PCR was used to detect the expression of?-catenin at the RNA level,immunofluorescence was used to detect the nuclear translocation of?-catenin in colorectal cancer cells,and cell scratch test and transwell chamber method were used to test the migration and invasion of colorectal cancer cells.Results:PRR11 was located in the cytoplasm/cell membrane of colorectal cancer.In colorectal cancer,PRR11 was highly expressed in colorectal cancer.The positive expression rate of cancer group was 60.92%(82/136),and that of adjacent group was 1.47%(1/68).There was a significant difference between cancer group and adjacent group(x~2=65.000,P<0.01).The expression of PRR11 in colorectal cancer was correlated with its clinicopathological factors,including distant metastasis,tumor size and tumor stage(P<0.05),but not with age,gender,tumor differentiation,tumor invasion depth and lymph node metastasis(P>0.05).The ectopic expression rate of?-catenin was 69.85%(95/136)in colorectal cancer and 0%(0/68)in adjacent tissues,and there was significant difference between the two groups(x~2=88.899,P<0.01).The ectopic expression rate of?-catenin was correlated with tumor differentiation,distant metastasis,tumor size and stage(P<0.05),but not with age,gender,tumor invasion depth and lymph node metastasis(P>0.05).The expression of E-cadherin was mainly located in the cell membrane.The deletion rate of E-cadherin was 76.47%(104/136)in colorectal cancer and 0%(0/68)in paracancerous tissue,and the difference between the two groups was statistically significant(x~2=106.080,P<0.01).The loss rate of E-cadherin was related to the degree of tumor differentiation,lymph node metastasis and stage(P<0.05),but not to age,gender,depth of tumor invasion,distant metastasis and tumor size(P>0.05).The expression rate of vimentin was 31.62%(43/136)in colorectal cancer and 2.94%(2/68)in adjacent tissues,and there was a significant difference between the two groups(x~2=21.683,P<0.01).The expression of vimentin was related to distant metastasis and stage(P<0.05),but not to age,sex,tumor differentiation,tumor invasion depth,lymph node metastasis and tumor size(P>0.05).PRR11 was highly expressed in colorectal cancer cells(SW480,HT29,HCT116)at RNA level and protein level,but low in normal colorectal epithelial cells(HCoEpiC),and the difference was statistically significant(P<0.05).The results of RT-PCR showed that PRR11was the most expressed in HT29 cells at RNA level(P<0.05),and PRR11 was the most expressed in SW480 and HT29 cells at protein level(P<0.05).The overexpression and silencing model of PRR11 was established by HT29 cells with good growth and relatively high PRR11 expression.The results showed that the expression of PRR11 in HT29 cells in overexpression group was up-regulated(P<0.05),and the ability of cell migration and invasion in the overexpression group was significantly increased(P<0.05).The expression of PRR11 in HT29 cells in silencing group was down regulated(P<0.05),and the ability of cell migration and invasion in the silencing group was significantly decreased(P<0.05).Western blot,immunofluorescence and RT-PCR were used to study the effects of PRR11 overexpression and silencing on the expression of EMT related signal molecules(?-catenin,E-cadherin,vimentin)at protein level and RNA level respectively.The results showed that?-catenin and vimentin in PRR11 overexpression group were significantly up-regulated,E-cadherin was significantly down regulated(P<0.05),and?-catenin had significantly nuclear translocated in cells.While in PRR11 silence group,?-catenin and vimentin were significantly down regulated,E-cadherin was significantly up-regulated(P<0.05),and?-catenin had not significantly nuclear translocated in cells.At the RNA level,the expression of?-catenin in PRR11 overexpression group was significantly up-regulated compared with the control group,while the expression of?-catenin in PRR11 silence group was significantly down regulated compared with the control group(P<0.05).Conclusion:PRR11 is highly expressed in colorectal cancer,and has a certain relationship with invasion and metastasis of colorectal cancer.PRR11may promote the progression of colorectal cancer by regulating Wnt/?-catenin signal pathway and promoting?-catenin nuclear translocation.