Study On The Role Of NOX1 In The Pathogenesis And Development Of Colorectal Cancer

Objective:To clarify the expression of NADPH oxidase 1(NOX1)in colorectal cancer and its relationship with clinical characteristics.To explore whether NOX1 can cause the proliferation and migration of colorectal cancer cells through oxidative stress imbalance,and promote the development of colorectal cancer,which provides experimental basis for the clinical diagnosis and treatment of colorectal cancer patients.Method:From January 2019 to December 2020,91 cases of patients with primary colorectal cancer were collected,and the expression of NOX1 protein in human colorectal cancer tissue was detected by immunohistochemistry(IHC).Small interfering RNA(siRNA)was used to interfere with the expression of NOX1 in human colorectal cancer cell line(DLD1).The expression of NOX1 mRNA in DLD1 cells was detected by fluorescence quantitative PCR.The expression level of NOX1 protein in DLD1 cells was detected by Western blotting.CCK-8 assay was used to detect the growth and proliferation of DLD1 cells.Transwell chamber was used to evaluate the migration ability of DLD1 cells.The content of reactive oxygen species(ROS)in DLD1 cells was detected by fluorescent probe DCFH-DA.Result:1.The results of immunohistochemistry showed that the expression of NOX1 was increased in human colorectal cancer tissues.According to different clinical indicators,the expression differences of NOX1 in each group were statistically analyzed.The results showed that the expression of NOX1 was related to the degree of tumor differentiation,tumor diameter and distant metastasis(P<0.05),but was related to gender,age,tumor location,vascular and nerve invasion,and lymph node metastasis,there was no significant correlation with metastasis(P>0.05).2.The results of fluorescence quantitative PCR and Western blotting showed that the mRNA and protein levels of NOX1 siRNA intervention were significantly lower than those of negative control group(NC)and blank control group(P<0.05);there was no significant difference in the expression level of NOX1 between NC group and blank control group(P>0.05).3.CCK-8 results showed that compared with NC group and blank group,there was no difference in cell proliferation level and proliferation rate of NOX1 siRNA intervention group(P> 0.05)The cell proliferation and cell proliferation control rate of siRNA intervention for 48 h,72h and 96 h were significantly decreased(P<0.05).There was no significant difference between NC group and blank group(P>0.05).4.Transwell migration experiment showed that the number of cell migration in NOX1 siRNA intervention group was significantly lower than that in NC and blank groups(P<0.05),but there was no significant difference between NC group and blank group(P>0.05).5.Fluorescence probe DCFH-DA was used to measure the content of ROS in DLD1 cells.The results showed that the level of ROS in NOX1 siRNA intervention group was lower than that in NC and blank group,and the difference was statistically significant(P<0.05).There was no significant difference between NC group and blank group(P>0.05).Conclusion:1.This study confirmed that the expression of NOX1 was increased in colorectal cancer tissues,and its expression was related to tumor differentiation,tumor diameter and distant metastasis,suggesting that the high expression of NOX1 may be involved in the pathogenesis of colorectal cancer.2.Knockout of NOX1 expression in human colorectal cancer cell line(DLD1)significantly inhibited the proliferation and migration of DLD1 cells,and knockdown of NOX1 expression inhibited the generation of ROS in DLD1 cells,suggesting that NOX1,as one of the sources of ROS generation,may participate in the pathogenesis and development of rectal cancer through mediating oxidative stress.Therefore,NOX1 is expected to become a target for biological therapy of colorectal cancer.