C19orf10 Promotes Malignant Behaviors Of Human Bladder Carcinoma Cells Via The PI3K/AKT Pathway

Bladder cancer(BC)is a common malignant tumor of the genitourinary system.In China,the incidence and mortality of BC rank first among the malignant tumors of the male genitourinary system.Therefore,it is necessary to get a better understanding of the molecular mechanism in BC development,so that we can improve the prognosis and effect of treatment.The protein encoded by the Chromosome 19 open reading frame 10(C19orf10)gene is a paracrine protein secreted by monocyte and macrophage.By consulting the literature,it is found that there are few studies on C19orf10,mainly focusing on acute myocardial infarction.Synthetic C19orf10 has been used to treat acute myocardial infarction(AMI)in mice,which can significantly reduce the area of ventricular infarction,improve ventricular remodeling and systolic dysfunction,and benefit the treatment and prognosis of AMI.There are few studies on C19orf10 in tumorigenesis and development.It has been reported that overexpression of C19orf10 promotes the proliferation of liver cancer cells,directly affects the self-renewal of liver cancer stem cell,and indirectly aggravates the inflammatory microenvironment to accelerate the development of liver cancer.However,its function and clinical value in BC have not been reported yet.This study aims to investigate the expression and function of C19orf10 in BC and explore its underlying mechanism.The main research contents and results are as follows:(1)Expression of C19orf10 and its relationship with clinicopathological characteristics of BCThe expression of C19orf10 in human BC tissues and cell lines was analyzed by reverse transcription-quantitative polymerase chain reaction(RT-q PCR)and Western blot.The results showed that the expression of C19orf10 in BC tissues and cell lines was significantly up-regulated compared with adjacent tissues and normal bladder urothelial cells.BC tissue chip was used to analyze the correlation between the protein level of C19orf10 and patient’s clinicopathological characteristics,and it showed that the expression of C19orf10 was related to the histological grade of tumor,suggesting that C19orf10 has a certain relationship with the malignant degree of BC,and C19orf10 may be useful for the diagnosis of BC.(2)The effect of C19orf10 on the malignant behaviors of BC cellsC19orf10 silence was achieved by small interfering RNA(si RNA)transfection and C19orf10 overexpression was obtained by plasmid transfection.Cell proliferation was detected by CCK-8,colony forming assay,and Ed U,while cell migration and invasion were detected by Transwell.The results showed that silencing C19orf10 significantly inhibited bladder cancer cell proliferation,migration and invasion.The overexpression of C19orf10 promoted cell proliferation,migration and invasion.A xenograft mouse model was used to detect the effect of small hairpin RNA(sh RNA)mediated stable silence of C19orf10 on the tumor-forming ability of BC cells in vivo.The results showed that compared with control mice,silencing C19orf10 significantly slowed down the tumor growth in xenograft mice,decreased the tumor volume,and reduced the tumor weight.The markers of proliferating cells including Ki67(MKI67)and PCNA(Proliferating cell nuclear antigen)were detected by immunohistochemistry,and it demonstrated that the number of Ki67 and PCNA positive cells was significantly decreased by C19orf10 silencing.These results indicate that silencing C19orf10 can inhibit tumor growth in xenograft mice,suggesting that C19orf10 may has a cancer-promoting effect.(3)The role of PI3K/AKT signaling pathway in C19orf10-driven malignant behaviors of BC cellsThe components of PI3K/AKT(Phosphatidylinositide 3-kinase/Protein kinase B)signaling pathway were examined by Western blot.The results showed that compared with the control group,C19orf10 silence led to a decrease in the protein levels of PI3 K and p-AKT,as well as an increase in the expression of AKT targets,including the expression of cell cycle inhibitors p21 and p27.Overexpression of C19orf10 resulted in the opposite.Furthermore,treatment with SC-79,a compound that can induce AKT phosphorylation,significantly increased the proliferation,migration and invasion of BC cells with C19orf10 silence comopared to those of the control group,indicating that upregulated AKT phosphorylation by SC-79 can restore the inhibitory effects of silencing C19orf10,which suggests that C19orf10 promotes the malignant behaviors of BC cells through the PI3K/AKT signaling pathway.In summary,C19orf10 expression is elevated in BC,which promotes the proliferation and metastasis of BC cells through the PI3K/AKT signaling pathway,suggesting that C19orf10 is a potential BC oncogene.This study provides new clues for the targeted therapy of BC.