Effect Of CNIH4 On Glioma Progression By The PI3K/Akt Signaling Pathway

Objective:By studying the expression of CNIH4 in glioma and its relationship with the prognosis of patients,the influence of CNIH4 expression on glioma related biological function was analyzed.Finally,the relationship between CNIH4 and PI3K/Akt signaling pathway in glioma progression was discussed and verified.Methods:1.The transcriptome data and clinical data of glioma samples were downloaded from the Chinese glioma Genome Atlas Project(CGGA)database,and the data on CNIH4 expression,survival rate and clinical characteristics of glioma patients from the Cancer Genome Atlas Project(TCGA)database were downloaded and applied for verification.2.Detected the expression of CNIH4 in glioma tissue and its para-carcinoma tissue samples by q RT-PCR and Western blot.3.Detected the expression of CNIH4 in NHA,LGG cell lines(SW1088,SW1783,BT142)and HGG cell lines(U87,U251,and LN229)by q RT-PCR and Western blot.4.Selected the cell line with suitable expression of CNIH4 to construct the cell model of knockdown CNIH4.Western blot and q RT-PCR were used to verify and detect the difference of CNIH4 expression levels.5.Used Cell Counting Kit-8,5-ethynyl-29-deoxyuridine,transwell and wound healing experiments,to detect the effects of CNIH4 knockdown on the proliferation,migration and invasion of U251 and LN229 glioma cells.6.The expression levels of PI3 K and Akt in knockdown CNIH4 cells were detected by Western blot.Results:1.The expression of CNIH4 is increased in glioma,and the higher the CNIH4 gene expression,the worse the prognosis.Univariate and multivariate Cox regression analysis showed that CNIH4 expression was an independent prognostic factor in glioma patients.We also found that CNIH4 expression was closely related to immune-related signals,immune cell infiltration,immune response regulation and chemotherapy response in glioma patients.2.The results of q RT-PCR and Western blot showed that the expression of CNIH4 protein in glioma patients was significantly higher than that in paracarcinoma tissues(P < 0.05).The CNIH4 m RNA of cancer tissue was significantly higher than that of para-carcinoma tissues(P < 0.01).3.The results of q RT-PCR and Western blot showed that the expression of CNIH4 protein in many glioma cell lines was significantly higher than that in normal astrocytes(NHA),and the difference was statistically significant.The CNIH4 m RNA of glioma cells was significantly higher than that of NHA,and the difference was statistically significant.The differential expression was more obvious in U251 and LN229 glioma cell lines.4.The proliferation ability of CNIH4 in glioma cells was significantly inhibited after the expression of CNIH4 was down-regulated(p<0.01).Similarly,the ability of metastasis and invasion of glioma cells decreased significantly(p<0.001).5.After down-regulating the expression of CNIH4 in glioma cells,the expression levels of p-PI3 K and p-Akt proteins in glioma cells decreased significantly(P<0.01).Conclusion:In conclusion,this study found that CNIH4 is highly expressed in gliomas,and confirmed that CNIH4 plays a series of cancer-promoting roles through the PI3K/Akt signaling pathway.We believe that this study can not only provide a new idea for revealing the pathogenesis and progression of glioma,but also provide a candidate target molecule for the study of new glioma targeting drugs for the benefit of glioma patients.