The Effect And Mechanism Of SLERT On Metastasis And Invasion Of Endmetrial Carcinoma

BackgroundEC is one of the main types of malignant tumor of female reproductive system.At present,the incidence of EC is increasing year by year and shows a trend of younger age.In some western countries and some economically developed areas of our country,the incidence of EC even exceeds that of cervical cancer,becoming the most common reproductive system malignant tumor affecting women.Although most patients with EC have a good prognosis,the mortality rate in advanced EC is high,especially when metastasis occurs.Activation of BDNF/Trk B signaling pathway can promote EMT and increase EC invasion and metastasis.Recent studies have shown that a novel Lnc RNA named SLERT plays a key role in gene regulation and may be related to the occurrence and development of tumors.Previous experimental studies have confirmed that SLERT is highly expressed in EC tumor tissues and plasma of EC patients,and is significantly correlated with FIGO stage,invasion depth,lymphatic vascular invasion and lymphatic metastasis of EC,which suggests that SLERT may be related to the occurrence and development of EC.Here,we explore the effects of SLERT on EC metastasis and invasion.ObjectiveBy investigating the role of SLERT in EC,this experiment is expected to reveal the molecular mechanism of EC metastasis and invasion from a new perspective,providing a theoretical basis for the clinical diagnosis,treatment,prediction and prognosis of EC.Method1.We collected survival information from 92 patients and analyzed the relationship between SLERT expression and survival using SPSS 22.0.We also analyzed the correlation between SLERT expression levels and clinical features such as tumor stage,grade,and lymph node metastasis.2.Total RNA was extracted from EC cells using Trizol reagent,followed by c DNA synthesis and gene amplification and quantification using q RT-PCR.KLE and AN3 CA cells infected with SLERT sh RNA and control cells were cultured in medium containing 2μg/ml puromycin,and stable SLERT sh RNA-expressing cell lines were screened for subsequent experiments.To investigate the effect of SLERT on the biological functions of EC cells,the CCK-8 assay kit was used to detect cell proliferation and evaluate the effect of SLERT on in vitro proliferation of EC cells.The cell scratch test was used to investigate the effect of SLERT on the in vitro migration ability of EC cells.Furthermore,the Transwell cell invasion assay was used to study the effect of SLERT on the in vitro invasion of EC cells.3.Western blotting will be used to detect the expression of BDNF,Trk B,and EMT-related proteins in EC cells with SLERT knockout or high expression.The correlation between SLERT and E-cadherin,Ncadherin,Vimentin,and BDNF will be analyzed using the TCGA database to verify the experimental hypothesis.4.The BDNF promoter will be synthesized and inserted into the PGL3-basic vector.Lipofectamine3000 will be used to transfect 293 T,KLE,and AN3 CA cells,and luciferase reporter assays will be performed to detect promoter activity and explore the interaction between SLERT and BDNF m RNA.Result1.After statistical analysis,it was found that SLERT was positively correlated with FIGO stage,depth of invasion,lymphovascular invasion,and lymph node metastasis,indicating that high expression of SLERT is associated with poor prognosis in EC patients.2.CCK-8 experiments showed that downregulation of SLERT did not affect cell viability,while scratch and Transwell assays showed that downregulation of SLERT weakened the migration and invasion of EC cells.3.Detection of EMT-related protein levels in KLE and AN3 CA cells after SLERT knockdown showed a negative correlation between SLERT and E-cadherin,and a positive correlation between SLERT and Ncadherin and Vimentin,suggesting that SLERT knockdown may inhibit the EMT process in EC cells.4.Analysis of the TCGA-EC database showed a significant negative correlation between SLERT and Ecadherin,and a significant positive correlation between SLERT and Vimentin,N-cadherin,and BDNF,suggesting that SLERT may promote EC cell migration and invasion by controlling EMT through BDNF.5.The dual-luciferase reporter assay showed that knocking down SLERT had no effect on BDNF promoter activity,but after treatment with actinomycin D,the levels of BDNF m RNA were significantly reduced in KLE and AN3 CA cells with SLERT knockdown,indicating that SLERT may inhibit the degradation of BDNF m RNA.On the other hand,overexpression of SLERT resulted in an increase in BDNF m RNA expression levels,suggesting that SLERT may affect the transcription process of BDNF m RNA.6.After overexpression of SLERT resulted in a decrease in E-cadherin levels,an increase in BDNF and its downstream Trk B protein levels,enhanced invasion ability in KLE and AN3 CA cells,and a decrease in invasion ability after BDNF knockout.These findings suggest that SLERT induces EMT and enhances EC cell invasion ability by regulating the BDNF/Trk B signaling pathway.ConclusionSLERT,as a lnc RNA promoting metastasis,may promote EMT process by increasing BDNF m RNA and activating BDNF/Trk B signal transduction pathway,thus enhancing the invasion and metastasis ability of EC cells.These results provide important clues to further explore the role of SLERT in the occurrence and development of EC,and provide experimental reference for finding new therapeutic targets and clinical intervention.