Expression And Significance Of KIAA1199 In Non-Small Cell Lung Cancer And Its Regulation Effect On Non-Small Cell Lung Cancer Cells

Background and Objective:KIAA1199 gene is closely related to the occurrence and development of many kinds of tumors.The expression,function and mechanism of KIAA1199 in non-small cell lung cancer(NSCLC)have not been clarified.Therefore,the purpose of this study was to analyze the expression of KIAA1199 in non-small cell lung cancer(NSCLC)and its correlation with the clinical features of patients with non-small cell lung cancer(NSCLC).To further study the relationship between KIAA1199 expression and prognosis in patients with non-small cell lung cancer(NSCLC).At the same time,the relationship between KIAA1199 gene and proliferation,apoptosis,migration and invasion of non-small cell lung cancer cells was studied,to study the effect of KIAA1199 gene on Wnt pathway,and non-small cell lung cancer cells were used to establish an animal model of nude mouse lung cancer,and further determine the role and significance of KIAA1199 in non-small cell lung cancer.Preliminarily Investigated the mechanism of action of KIAA1199 gene in non-small cell lung cancer cells.Methods:The expression of KIAA1199 in lung cancer tissues was analyzed by immunohistochemistry.The correlation between protein expression of KIAA1199 and clinicopathological factors was analyzed.Univariate survival curves and multivariate Cox regression were used to analyze the potential prognostic factors of NSCLC patients.Normal lung epithelial cells(BEAS-2B)non-small cell lung cancer cell lines(NCI-H460,A549,NCI-H1299,SK-MES-1)were cultured in vitro to detect the expression of KIAA1199 gene and protein in cell lines of each group.To detect the effect of KIAA1199 knockout and overexpression on proliferation,apoptosis,migration and invasion of NSCLC cells.The effect of KIAA1199 on Wnt signaling pathway activity and expression of β-Catenin protein was detected,and the expression of Wnt signaling pathway related genes in non-small cell lung cancer cells after KIAA1199 gene knockout was detected.Non-small cell lung cancer cells were used to establish an animal model of lung cancer in nude mice.To detect the effect of KIAA1199 interference on tumor cell proliferation and apoptosis in nude mice.Results:1.The expression of KIAA1199 protein in 76 cases(49.67%)increased in NSCLC patients,Statistical analysis showed that KIAA1199 protein expression was up-regulated in NSCLS tissues than normal(P=0.003)2.High expression of KIAA1199 protein was correlated with differentiation,pathological type,lymph node metastasis and TNM stage in NSCLC lung cancer(P<0.05).But the degree of KIAA1199 protein expression was not related to gender and age in patients with NSCLC(P>0.05).3.The progression-free survival(PFS)and overall survival(OS)of NSCLC patients were not affected by gender and age factors,and were not related to pathological types of patients(P>0.05).Is related to tumor differentiation,protein expression of KIAA1199,lymph node metastasis and TNM stage in NSCLC patients(P<0.05).4.Multivariate analysis showed that the expression of KIAA1199 protein was an independent prognostic factor of PFS in NSCLC patients(HR=0.661,P = 0.027).It is also an independent prognostic factor of OS in NSCLC patients(HR=0.711,P =0.041).5.Patients with NSCLC were divided into groups by the pathological type.The overexpression of KIAA1199 protein was associated with the prognosis of NSCLC patients aged less than 60 years.Prognosis of NSCLC patients with positive lymph node metastasis or advanced stage were affected by expression of KIAA1199.6.Compared with the normal bronchial epithelial cell line(BEAS-2B),the gene expression of KIAAl199 increased in the non-small cell lung cancer cell lines of each group,and the difference was statistically significant(P<0.05).7.After knockdown the expression of KIAA1199 in H460 and A549 cells,the proliferation rate of non-small cell lung cancer cells was significantly decreased,the apoptosis of non-small cell lung cancer cells was promoted,and the invasion and migration of non-small cell lung cancer cells were inhibited.The overexpression of KIAA1199 in H1299 and SK-MES-1 cells could significantly increase the proliferation rate of non-small cell lung cancer cells,inhibit the apoptosis of non-small cell lung cancer cells,and enhance the invasion and migration of non-small cell lung cancer cells.8.The activity of Wnt signaling pathway decreased after knockdown of KIAA1199 expression.The activity of Wnt signaling pathway could be restored by adding Wnt signaling pathway agonist(SKL2001).After overexpression of KIAA1199,the activity of Wnt signaling pathway increased.The addition of Wnt signaling pathway inhibitor(FH535)could inhibit the activity of Wnt signaling pathway,and overexpression of KIAA1199 could not reverse the inhibitory effect of FH535.9.Overexpression or knockdown of KIAA1199 had no significant effect on the total content of β-Catenin protein in lung cancer cells.After overexpression of KIAA1199,the content of β-Catenin protein increased in the nucleus and decreased in the cytoplasm.After knocking down KIAA1199,the content of-catenin protein in the nucleus decreased and increased in the cytoplasm.10.Knockdown of the expression of KIAA1199 in A549 cells can significantly reduce the expression of c-myc,CyclinDl and CD44 in non-small cell lung cancer cells.The expression of c-myc,CyclinDl and CD44 could be recovered by adding Wnt signaling agonist(SKL2001).11.KIAA1199shRNA transfection in nude mice with non-small cell lung cancer could delay the formation of tumor and significantly reduce the diameter of tumor.12.KIAA1199 knockdown significantly reduced the expression of ki-67 protein in tumor tissues of nude mice,and apoptotic cells were significantly increased.Conclusion:1.KIAA1199 gene overexpression was found in NSCLC tissues and cell lines.2.The expression level of KIAA1199 protein was correlated with the pathologic type,histological differentiation,lymph node metastasis and TNM stage in patients with NSCLC3.KIAA1199 protein expression is related to progression-free survival and overall survival of NSCLC patients,and is an independent prognostic factor for progression-free survival and overall survival of NSCLC patients.4.KIAA1199 can participate in the regulation of NSCLC cell proliferation,apoptosis,migration and invasion.5.KIAA1199 regulates the activity of Wnt signaling pathway and the expression of downstream target molecules by regulating the translocation of β-Catenin expression.6.Knockdown of KIAA1199 can inhibit the tumor progress of NSCLC animal model.Inhibit the proliferation of tumor cells in nude mice and promote the apoptosis of tumor cells in nude mice.