Expression Of WNK1 In Colorectal Cancer And Its Influence On Prognosis

Background:The incidence of colorectal cancer(CRC)from high to low is rectum,sigmoid colon,cecum,ascending colon,descending colon and transverse colon,which is a relatively common gastrointestinal malignant tumor.The incidence of colorectal cancer is closely related to lifestyle,eating habits,genetics,etc.,and as people's lifestyle changes and quality of life improves,the incidence of colorectal cancer is getting younger and younger.Due to the intestinal anatomy and functional characteristics,early colorectal cancer is often atypical or asymptomatic,and it is easy to be overlooked.The patient is relatively late in the disease when it is diagnosed.Even patients who can be treated with surgery should receive further chemotherapy and biological Treatment,etc.,has brought great financial burden and mental pressure to patients and their families.Even if CRC patients are actively treated,due to the occurrence of recurrence and metastasis after surgery,drug resistance,etc.,most CRC patients have poor overall prognosis and poor quality of life.In order to diagnose or evaluate CRC,tumor markers commonly used in clinical work include carcinoembryonic antigen(CEA)and carbohydrate antigen(CA19-9),but the specificity of these tumor markers is not ideal,and the indicators are susceptible to many other Due to various physical factors,the function of disease prediction and evaluation is limited.Therefore,it is urgent to find new tumor markers and more sensitive indicators to evaluate the prognosis of patients,so as to find new targets and directions for the diagnosis and treatment of CRC patients.WNK1[lysine-free(K)-1]is a 250 kDa serine/threonine protein kinase,which is widely expressed in the body,and more research is currently performed on WNK1 expressed in the nephron of the collecting duct.Studies have found that WNK1 plays an important role in maintaining the balance of water and electrolytes in cells,and is involved in the occurrence and development of hereditary hypertension.WNK1 can regulate the functions of different cells and organs through a variety of mechanisms,and plays a role in various pathophysiological processes.MONIZ S and others have found that WNK1 has an important effect in neoplastic diseases,mainly through regulating tumor cell proliferation,metabolic adaptation,apoptosis escape,invasion and metastasis.Down-regulation of WNK1 and WNK2 in HEK293T cells can significantly reduce the expression of developing core Wnt signaling genes,indicating that knocking out WNK1 and WNK2 genes can reduce the activity of HEK293T cells.Vegf/Vegfr signaling pathway can regulate angiogenesis,the mechanism is through the regulation of Akt kinase-mediated phosphorylation and WNK1 activation.ZHANG et al.found that WNK1,as the second cytoplasmic messenger,articipates in the Cl-channel-induced sipnalingg cascade and romotes angiogenesis when studying the related influencing factors of angiogenesis.The formation of new blood vessels is very important for tumor growth and metastasis.Combined with previous studies on WNK1,the expression of WNK1 gene may have a certain effect on the aggressive behavior of tumor cells.According to reports,the copy number of the WNK1 gene in CRC is abnormal,but the specific impact of WNK1 on the clinical characteristics of CRC and its mechanism are still unclear.In this study,by examining the expression of WNK1 in CRC,the relationship between WNK1 and clinicopathological characteristics and prognostic indicators of CRC patients was analyzed,and the mechanism of WNK1 on CRC cells was further studied to clarify the effect of WNK1 on CRC.Objective:To study the expression of WNK1 in CRC tissue and its adjacent tissues and its effect on the migration of CRC cells;to analyze the effect of WNK1 on the clinical effect indexes of CRC patients.Method:1.The 68 CRC tissue samples used in this research were provided by the specimen bank organized by Shanghai Xinchao Biotechnology Co.,Ltd.According to the eighth edition of TNM staging criteria for colorectal cancer,the cases were clinically pathologically staged.There were 41 males and 27 females,with an average age of 55 years.Each research specimen provides cancer tissue and adjacent tissue,and the adjacent tissue is 1.5 cm away from the cancer tissue.The expression of WNK1 gene in CRC tissue and its adjacent tissues was detected by immunohistochemistry.2.Data analysis is performed with SPSS 21.0 statistical software.?2 test was used to evaluate the relationship between the expression of WNK1 in cancer tissues and adjacent tissues and the relationship between the expression of WNK1 and the clinicopathological features of CRC.Log-rank test and Kaplan-Meier survival curve were used to analyze the relationship between WNK1 expression level and overall survival(OS)and progression-free survival(PFS).The measurement data is indicated by(x±s),and the independent sample t test is used for comparison between the two groups.Setting P<0.05 means that the difference is obvious and statistically significant.3.Evaluate the effect of WNK1 on the migration ability of CRC cells through Scratch test and Transwell test.Result:1.The expression of WNK1 in CRC cancer tissues and adjacent tissues,and the correlation analysis between WNK1 expression and the clinicopathological characteristics of CRC patients showed that the positive expression rate of WNK1 in CRC tissues[(55.9%(38/68)]was higher than that in CRC tissues[14.7%(10/68)],with statistically significant difference(P<0.05).The expression of WNK1 was correlated with the number of lymph node metastasis and TNM stage in CRC patients(all P<0.05).WNK1 expression was not correlated with age,sex,tumor diameter and tumor infiltration depth of CRC patients(all P>0.05).2.Log-rank test and Kaplan-Meier survival curve analysis showed that the overall survival rate of colorectal cancer patients with high WNK1 expression was lower than that of patients with low WNK1 expression(P<0.05),and the difference was statistically significant.The progression-free survival rate of colorectal cancer patients with high WNK1 expression was lower than that of patients with low WNK1 expression(P<0.05),and the difference was statistically significant.3.Transwell results:In HCT116 cells,the number of WNK1 gene inhibited by siRNA and migrated to the lower compartment through the membrane was(57±4.36)less than that of the empty vector control group(230±8.89),with statistically significant difference(P<0.05).In HCT15 cells,the number of WNK1 gene inhibited by siRNA and migrated to the lower compartment through the membrane was(61±3.61)less than that of the empty vector control group(241±5.52),and the difference was statistically significant(P<0.05).4.Scratch test results:The scratch healing rate(10.7%)in HCT116 cells 48 h after the WNK1 gene was down-regulated by siRNA was lower than that in the empty vector control group(60.3%),with statistically significant difference(P<0.05).After the WNK1 gene was down-regulated by siRNA in HCT115 cells at 48 h,the wound healing rate(7.8%)was lower than that of the empty vector control group(45.8%),the difference was statistically significant(P<0.05).Conclusion:1.WNK1 expression in CRC tissues was significantly higher than that in paracancer tissues;WNK1 expression was significantly correlated with advanced TNM stage and lymph node metastasis in CRC patients,suggesting that WNK1 may be an indicator of poor prognosis in CRC patients.2.The OS and PFS of CRC patients with high WNK1 expression were shorter,suggesting that the prognosis of CRC patients with high WNK1 expression was poor.3.Inhibition of WNK1 gene can inhibit the migration of CRC cells,and WNK1 may play a pro-oncogenic role in CRC.