APOBEC1 Complementation Factor Facilitates Cell Migration By Promoting Nucleus Translocation Of SMAD3 In Renal Cell Carcinoma Cells And High Dose Dexamethasone Injection Dysregulated Multiple-protein Kinases Expression In The Mouse Kidney

Metastasis is inevitable in about 30% of patients with primary clear cell renal cell carcinoma after nephrectomy treatment.Exploring molecular mechanism of renal cell carcinoma metastasis conduces to developing new therapeutic targets and improving prognosis.A1 CF,an RNA binding protein,participates in tumor progressions such as growth,apoptosis,differentiation,and invasion.However,it remains unclear whether A1 CF exerts an effect on migration in renal cell carcinoma cells.Here,we explored effects of A1 CF in clear cell renal cell carcinoma.A1 CF overexpression and knockdown stable cell lines were constructed by lentiviral transfection.Wound healing assay was conducted to detect migration in stable cell lines.Quantitative PCR and Western blot assays were utilized to test transcriptional and translation levels of A1 CF and SMAD3 in A1 CF overexpression and knockdown stable cell lines.Nuclear and cytoplasmic protein separation assays were conducted to evaluate the subcellular distribution of A1 CF and SMAD3.Immunoprecipitation assay was conducted to detect the interaction between A1 CF and SMAD3 in HEK293 T cells transfected with A1 CF and SMAD3 overexpression plasmids.Rescue assay was performed to confirm whether SMAD3 participated in cell migration induced by A1 CF overexpression.Our study demonstrated A1 CF overexpression facilitated cell migration in 786-O renal carcinoma cells.SMAD3 expression is positively correlated with A1 CF in renal carcinoma from TCGA database.Elevated expression of A1 CF raised SMAD3 expression and depletion of A1 CF correspondingly downregulated SMAD3 expression in transcriptional and translation levels.A1 CF deficiency downregulated expression of Snail1 and N-cadherin.In addition,A1 CF promoted nucleus translocation of SMAD3 and interacted with SMAD3.SMAD3 knockdown attenuated cell migration induced by A1 CF overexpression.Our study suggested A1 CF facilitated cell migration by promoting nucleus translocation of SMAD3 in renal cell carcinoma cells.Glucocorticoids have been used to treat inflammatory and immune disorders and shock in clinical cure.However,the effect of Glucocorticoids in immune response and metabolism of mice kidneys remains unclear.Here,we profiled the gene expression of kidney from mice with dexamethasone treatment via RNA-seq.Gene Ontology and KEGG pathway analysis showed ECM-receptor interaction,cell adhesion molecules signaling pathway were significantly enriched in differentially expressed genes,and that vast majority of differentially expressed genes were involved in monocarboxylic acid metabolic process,leukocyte cell-cell adhesion and fatty acid metabolic process.Furthermore,in differentially expressed genes,we found multiple protein kinase expression was dysregulated greatly in mice kidney after dexamethasone treatment.These protein kinases are involved in multiple signaling pathways in mice kidney,such as MAPK and PI3K/Akt signaling pathway.Our study provided important information for further study on the mechanism of side effect of glucocorticoids in clinical therapy.