The Mechanism Of LncRNA SNHG1 Interacting With HnRNPL To Promote The Progression Of Prostate Cancer

Background and purposeProstate cancer(PCa)is one of the common malignant cancer in urinary system.Recent studies revealed that LncRNA play critical roles in cancer progression.Small nuclear host gene 1(SNHG1),a novel lncRNA,was aberrantly expressed in multiple cancers and act as an oncogene.We previously discovered that SNHG1 is upregulated in PCa,however,the biological function and mechanisms of SNHG1 in PCa remained unknown.This study aims to further investigate the clinical relevance and functions of SNHG1 in PCa,and illustrate the detailed mechanisms.MethodsTCGA database was used to detect the expression and clinical significance of SNHG1;FISH was used to validate the SNHG1 expression and clinical relevance in 67 PCa and 14 normal prostate tissues.We constructed SNHG1 knockdown and overexpression PCa cell lines via lentiviral vector,CCK-8,cloning formation assays and EdU staining were subsequently used to detect the cell proliferation;Transwell migration assays and wound healing assays were utilized to detect the migration ability of PCa cells.Nude mice subcutaneous tumor formation test was used to detect the tumor formation ability.RNA sequence was used in the SNHG1 silencing PCa cells,and the specific molecules and pathways were validated by RT-qPCR and western blot.RNA pulldown combined with mass spectrometry was utilized to screen the potential proteins interacting with SNHG1,then bio-information prediction and RIP validated their binding.To further confirm the significance of SNHG1/hnRNPL complex,RIP assay was performed in SNHG1 silencing PCa cells after transfected with wild-type and binding site mutation SNHG1 overexpression plasmid.TCGA database was used to analyze the clinical relevance between SNHG1 and hnRNPL.Finally,hnRNPL silencing and mRNA stability assays were used to investigate the downstream molecules regulated by SNHGl/hnRNPL complex.ResultsSNHG1 was overexpressed in PCa tissues and cell lines,and positively correlated with Gleason grades,high expression of SNHG1 reduce the overall survival and disease-free survival.Knockdown of SNHG1 in PCa inhibited cell proliferation and migration ability in vitro,while overexpression showed the opposite effect.Besides,knockdown of SNHG1 reduced the in vivo tumorigenesis.RNA sequence revealed that SNHG1 silencing mainly affect the cell adhesion molecules(CAMs);RT-qPCR and western blot were validated that SNHG1 knockdown upregulates the expression of E-cadherin and inhibits the EMT progression in PCa,while SNHG1 overexpression showed the opposite effect.RNA pulldown and RIP assays validated the binding of SNHG1 and hnRNPL.Mutant SNHG1 could not bind to hnRNPL protein,hnRNPL also lost the ability to capture E-cadherin mRNA compared with wide-type SNHG1.TCGA database indicated that patients both overexpressing SNHG1 and hnRNPL have lower overall survival and disease-free survival.Finally,hnRNPL could stabilize the mRNA of E-cadherin.ConclusionSNHG1 is abnormally expressed in PCa,and its expression level is correlated with survival rate and progression in PCa.SNHG1 promotes the EMT process of PCa via competitively binding to hnRNPL,and inhibiting the expression of E-cadherin,leading to tumor proliferation and metastasis.Finally,this study demonstrates that SNHG1 and hnRNPL complex play a significant role in regulating the PCa progression,which could serve as a potential target for PCa therapy.