| Objective: To analyze the expression of ZNF281 in colorectal cancer and adjacent tissues and its correlation with pathological parameters and prognosis of patients;To explore the effect of silencing ZNF281 on 5-Fu chemotherapy sensitivity in colorectal cancer cells.Methods: Download and analyze the expression level of ZNF281 m RNA in colorectal cancer tissues and adjacent tissues from The Cancer Genome Atlas(TCGA)database;use RT-PCR to further verify ZNF281 in colorectal cancer tissues and adjacent normal tissues The expression level of ZNF281 m RNA;Western blot was used to further verify the expression level of ZNF281 protein in colorectal cancer tissues and matched normal tissues adjacent to the cancer of the center;ZNF281 expression in tumor tissue specimens of 150 colorectal cancer patients using immunohistochemistry Amount to be tested.At the same time,the clinical data of patients were collected,the correlation between ZNF281 and clinical pathological parameters was analyzed by statistical methods,survival curves were drawn,and Cox proportional hazards models were used to perform univariate and multivariate analysis of overall survival and relapse-free survival.Impact of prognosis in patients with colorectal cancer.RT-PCR and Western blot were used to detect the expression level of ZNF281 in colorectal cancer cells,and si RNA transient transfection method was used to down-regulate the expression level of ZNF281 in colorectal cancer cell lines.CCK-8experiment and flow cytometry were used to detect silent ZNF281 The effect of colorectal cancer cells on5-Fu chemotherapy sensitivity.Results: 1.Analysis of the downloaded TCGA data showed that ZNF281 showed high expression inunpaired colorectal cancer tissues,and low expression in normal tissues adjacent to cancer(P <0.001);compared to normal tissues adjacent to cancer,ZNF281 was found in matched colorectal tissues Cancer tissue also showed high expression(P <0.001);RT-PCR confirmed the high expression of ZNF281 m RNA in colorectal cancer tissues in our center;Western blot confirmed the high expression of ZNF281 in colorectal cancer tissues in our center.2.Immunohistochemical results showed that the positive and negative expression rates of ZNF281 in colorectal cancer tissue were 79.33%(119/150)and 20.67%(31/150),respectively,the difference was statistically significant(P <0.05).3.Clinical and pathological correlation analysis showed that the expression of ZNF281 was significantly correlated with tumor T stage,N stage,M stage,and TNM stage(P <0.05).Survival analysis showed that patients with high expression of ZNF281 had significantly shorter overall survival time and relapse-free survival time(P <0.05),and poor prognosis.COX regression analysis showed that: ZNF281 is an independent risk factor affecting the prognosis of patients with colorectal cancer.4.The results of CCK-8 experiment showed that the survival rate of colorectal cancer cells after silencing ZNF281 treated with 5-Fu was lower than that of the control group(P<0.05).5.Flow cytometry Annexin V / PI double staining method was used to detect apoptosis,and the apoptosis rate was compared: after 5-Fu treatment was used to silence ZNF281,the apoptosis rate was increased compared with the single-silence ZNF281 and 5-Fu treatment groups only There are statistical differences(P <0.01).6.Using flow cytometry to detect the cell cycle,knock down ZNF281.After 5-Fu treatment,the cell cycle changes were found to increase the proportion of G 1 + S phase cells,and the cell ratio was lowerthan that of single knockdown ZNF281 and 5-Fu treatment The group increased significantly with statistical difference(P <0.05).Conclusions: ZNF281 is up-regulated in colorectal cancer tissues,and ZNF281 expression is an independent risk factor that affects the prognosis of colorectal cancer patients;ZNF281 plays an important role in regulating the resistance of colorectal cancer cells and may provide a new molecular target for the precise treatment of colorectal cancer point. |
PDF Full Text Request