| Objective: To investigate the difference of the expression levels of RHBDD1,DLC1 andβ-catenin in colorectal cancer and para-carcinoma tissue,and to analyze the correlation of the expressions of RHBDD1,DLC1 and β-catenin in colorectal cancer and their relationship with clinicopathologic features.To elucidate the differences in the expression of CD3,CD8,PD-1and PD-L1 in the parenchyma and interstitium of colorectal cancer.To further understand the relationship between RHBDD1,DLC1,β-catenin and PD-L1 in colorectal cancer microenvironment.Method:1.qRT-PCR and western-blot were used to detect the expression of RHBDD1,DLC1 and β-catenin in colorectal cancer and para-carcinoma tissue,and spearman rank sum test was used to analyze the correlation of the expressions of Rhb DD1,DLC1 and β-Catenin in colorectal cancer and their relationship with clinicopathologic features.2.The differences in the expressions of CD3,CD8,PD-1 and PD-L1 in the parenchyma and interstitium of colorectal cancer were detected by multiple fluorescence immunohistochemistry.3.The expression of RHBDD1,DLC1,β-catenin and PD-L1 in colorectal cancer tissues were detected by immunohistochemistry,and the relationship between the three and PD-L1 expression in tumor tissue parenchyma and stroma were analyzed.Results:1.m RNA expression levels of RHBDD1,β-catenin and DLC1 in colorectal cancer and para-carcinoma tissue were(1.1783±0.1720)vs(1.0009±0.0005)(P<0.001),respectively;(1.3162±0.4262)vs(1.0010±0.0005)(P<0.001),(0.7919±0.2821)vs(1.0095±0.0060)(P<0.001);The expression levels of the three proteins in colorectal cancer and para-carcinoma tissue were(1.0497±0.2073)vs(0.81530.1637)(P<0.001),respectively.(1.1873±0.3667)vs(0.8533±0.2188)(P<0.001),(0.8933±0.1899)vs(1.1300±0.1992)(P<0.001);The expression levels of RHBDD1 and β-catenin in colorectal cancewere higher than those in para-carcinoma tissue.The expression level of DLC1 in colorectal cancer was lower than that in para-carcinoma tissue.2.m RNA and protein expressions of RHBDD1 and β-catenin in colorectal cancer were positively correlated(r=0.691,P<0.001),(r=0.679,P<0.001);The expression of RHBDD1 and DLC1 was negatively correlated(r=-0.655,P<0.001),(r=-0.743,P<0.001).There was a negative correlation between β-catenin and DLC1 expression(r=-0.776,P<0.001),(r=-0.656,P<0.001),with statistical difference.3.The expression levels of RHBDD1,DLC1 and β-catenin in colorectal cancer were not correlated with gender,age and tumor size(P>0.05).The expression levels of RHBDD1 and β-catenin were negatively correlated with the degree of tissue differentiation(r=-0.661,P<0.001),(r=-0.459,P=0.011).The expression level of DLC1 was positively correlated with the degree of tissue differentiation(r=0.653,P<0.001).The expression levels of RHBDD1 and β-catenin were positively correlated with TNM staging(r=0.644,P<0.001),(r=0.468,P=0.009).The expression level of DLC1 was negatively correlated with TNM staging(r=-0.461,P=0.01).The expression levels of RHBDD1 and β-catenin were positively correlated with lymph node metastasis(r=0.704,P<0.001),(r=0.587,P=0.001).The expression of DLC1 was negatively correlated with lymph node metastasis(r=-0.591,P=0.001).4.The expressions of CD3,CD8,PD-1 and PD-L1 in the tissue parenchyma of colorectal cancer were(8.3613±4.1383),(5.4683±2.7619),(1.0387±0.4838)and(8.5127±4.1616).The expressions in the interstitial of corresponding cancer were(12.7760±3.6597),(9.9123±5.3869),(1.3925±0.3762),(11.9420±5.4473),respectively.The expression of the patients in the parenchyma of colorectal cancer was lower than that in the interstitial of corresponding cancer,the difference was statistically significant(P<0.05).5.PD-L1 was positively correlated with the expression of RHBDD1 and β-catenin in the parenchyma and interstitium of colorectal cancer(r>0,P<0.05).PD-L1 expression was negatively correlated with DLC1 expression in tumor parenchyma and interstitium(r<0,P<0.05).Conclusion:1.The expression of RHBDD1 and β-catenin is abnormally increased in colorectal cancer,while the expression of DLC1 is decreased in colorectal cancer,suggesting that RHBDD1,DLC1 and β-catenin are involved in the occurrence of colorectal cancer,and the combined detection of the three may provide theoretical basis for the diagnosis of colorectal cancer.2.The expression of RHBDD1 was positively correlated with β-catenin in colorectal cancer.The expression of DLC1 was negatively correlated with β-catenin and RHBDD1.These results suggest that the interaction of the three factors in the occurrence and development of colorectal cancer is related to the progression and prognosis of the disease.3.The abnormal expressions of RHBDD1,DLC1 and β-catenin are correlated with the degree of tissue differentiation,TNM stage and lymph node metastasis of colorectal cancer,but not with gender,age and mass size.Therefore,the three expressions may affect the invasion,metastasis and prognosis of colorectal cancer.4.Higher expressions of CD3,CD8,PD-1 and PD-L1 in the interstitial tissue of colorectal cancer than in the parenchyma may lead to immunosuppression of the tumor microenvironment,thus affecting the anti-tumor effect of immune cells.5.RHBDD1,β-catenin were positively correlated with PD-L1 in parenchyma and stroma with abnormal expression of colorectal cancer tissues,and DLC1 was negatively correlated with PD-L1 in parenchyma and stroma,revealing that genes abnormally expressed in Wnt signaling pathway may participate in the regulation of tumor microenvironment in colorectal cancer. |
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