The Pseudogene DUXAP10 Promotes An Aggressive Phenotype Through Binding With LSD1 And Repressing LATS2 And RRAD In Non-small Cell Lung Cancer

Background:A recent study found that pseudogenes available to regulating the the homologous gene expression in transcription and transcription level,and participate in the process of cell biology,thus influence the proliferation,invasion and metastasis of tumor biological behavior.Pseudogenes are highly similar to encoding proteins genes,but cannot express of gene inside the cell,widely distributed in the genome of each species,particularly in most mammals.Pseudogenes DUXAP10 in 14 q11.2,the length is 2398 nt.Recent research reports,pseudogenes in tumor can exercise the function of oncogenes or tumor suppressor genes,however,DUXAP10 biological functions and the underlying mechanism in NSCLC is unclear.Objective:The aim of this study is to detect DUXAP10 expression in NSCLC and to evaluate the role and potential mechanisms of DUXAP10 in the development and progression of NSCLC.Methods:(1)The lncRNAs expression profiles of NSCLC patient from Gene Expression Omnibus(GEO)were analyzed.(2)Real-time quantitative PCR(qRT-PCR)assays was performed to detect the expression of DUXAP10 in NSCLC tissues and cells.(3)MTT and clone formation assays were used to evaluate the viability of NSCLC cells transfected with si-DUXAP10.(4)Transwell experiment found that after interference DUXAP10 expression can inhibit cell invasion and migration ability.(5)Nude mouse experiment was performed to observe the effect of A549 cell transfected with si-DUXAP10 on tumor ability in vivo.(6)QRT PCR and Western blot experiments show that after interference DUXAP10,the mRNA and protein expression level of LATS2 and RRAD are raised.(7)RIP and CHIP assays were performed to evaluate the potential mechanism that DUXAP10 how to regulate the downstream target gene.(8)Build LATS2 and RRAD expression plasmid,and by saving experiment DUXAP10 could regulate the expression of downstream target genes.Results:(1)Analysis of GEO database found that DUXAP10 was high expressed in NSCLC cancer organizations.(2)The results revealed that DUXAP10 expression was upregulated in NSCLC tissues and cells by qRT-PCR assay.Statistical analyses revealed that DUXAP10 expression levels in NSCLC were significantly correlated with tumor size,advanced pathological stage,and lymph node metastasis,also were correlated with poor prognosis of NSCLC.(3)Determined by MTT,clone forming experiments and EdU results,we found that interference DUXAP10 expression can inhibit tumor cell proliferation;Flow cytometry analysis revealed that knockdown of DUXAP10 expression induced cell cycle arrest at G1-G0 phase in NSCLC cells.(4)Transwell experiment found that after interference DUXAP10 expression can inhibit cell invasion and migration ability.(5)By building nude mice transplantation tumor model,we find that the inhibition of DUXAP10 expression can reduce A549 cells in vivo tumor ability.(6)qRT-PCR and Western blot experiments show that the after reducding DUXAP10,mRNA and protein expression level of LATS2 and RRAD are raised.(7)The RIP and Pulldown experiments showed that the enzyme LSD1 DUXAP10 can bind to methylated histones;ChIP experiments show that DUXAP10 could recruit LSD1 at promoter region of LATS2 and RRAD thus inhibiting LATS2 and RRAD transcription.(8)Rescuing experiment results show that raising expression of LATS2 and RRAD can save the effect of DUXAP10 promote proliferation.Conclusion:Our results firstly demonstrated that DUXAP10 contributed to NSCLC cell proliferation,partly though LSD1-medicated suppression of the LATS2 and RRAD.Our study may provide a novel strategy for targeting the DUXAP10/LSD1/LATS2,RRAD axis as a new therapeutic application for NSCLC patients.