Role Of LncRNA SChLAP1 Impacting On The Metastasis And Apoptosis Of Prostate Cancer And Its Mechanism

Research background and purpose:Prostate cancer(PC)is the first malignant tumor in men in western countries and the second most common malignancy in men worldwide.In China,the incidence of prostate cancer,with mainly diagnosed in advanced,is temporarily unable to compare with Western countries.Although there are many kinds of treatments,especially drugs,we still have a long way to go in the diagnosis and treatment of prostate cancer.With the constant development of molecular biology,long noncoding RNAs(LncRNAs)have involved various mechanisms in gene expression,such as regulation of transcription,translation,protein modification,and RNA-protein or protein-protein complexes.Especially in cancer,lncRNAs are also involved in regulating important cell signaling pathways.Although Lnc SChLAP1 was found to antagonize the effects of tumor suppressors,the specific mechanism of its action still needs further study.Methods:1.QPCR and fluorescence in situ hybridization detection of SChLAP1 in prostate cancer tissues.QPCR detection of SChLAP1 from PC-3,LNCaP,DU145 and WPMY-1 cell lines.2.Knockdown of SChLAPl by lentivirus,CCK-8 detection,Transwell assay,flow cytometric detection of apoptosis,flow cytometric detection of cell cycle and WB detection in prostate cancer cell lines3.The location of SChLAP1 in the localized cells was detected by fluorescence in situ hybridization and targeted miRNAs were searched by bioinformatics analysis and verified by dual luciferase targeting assays.4.Functional verification of target miRNAs in cell lines,obtaining ideal subjects through lentivirus overexpression and performing functional experiments including:CCK-8 assay,Transwell assay,flow cytometry(cell cycle and apoptosis),and WB Testing.Finally,in vivo verification was performed in nude mice.Results:1.The expression of SChLAPl in prostate cancer tissues was significantly higher than that in paracancerous tissues by qPCR(P<0.05).Fluorescence in situ hybridization showed that the tissues of prostate cancer was significantly higher than that in adjacent tissues,and the expression was correlated with Gleason score.There was a positive correlation,and PC-3 cells had the highest expression of SChLAP1 in cell lines.2.The expression of SChLAPl in PC-3 cells was reduced by transfection with lentivirus.After downregulation of SChLAPl in PC-3 cells,CCK-8 suggested that cell growth was significantly inhibited(P<0.05);Transwell assay showed that the invasive ability was significantly inhibited(P<0.001);flow cytometry suggested that the proportion of apoptosis was significantly increased(P<0.001);Flow cytometry suggested that the cell cycle was significantly blocked(P<0.01);WB results showed that knockdown of SChLAPl attenuated the expression of MMP-9,bcl-2 and caspase-3 without affecting the expression of E-cadherin.3.Fluorescence in situ hybridization results showed that SChLAP1 was mainly expressed in the cytoplasm of prostate cancer cells;bioinformatics detection showed that miR-101-5p was a possible target of SChLAPl;qPCR detection of PC-3 cells after lentivirus infection showed a clear mutual antagonistic relationship between them;dual luciferases show that the SChLAP1 3'UTR targets mir-101-5P directly.4.Upregulation of miR-101-5p expression in PC-3 cells by lentivirus transfection.After miR-101-5p was up-regulated in PC-3 cells,the growth of PC-3 cells was significantly inhibited by CCK-8(P<0.001);Transwell assay showed that the invasion ability was significantly inhibited(P<0.01);flow cytometry suggested that the cells withered.The percentage of death increased significantly(P<0.001);flow cytometry showed no significant changes in the cell cycle;inhibition of MMP-9,bcl-2 did not affect the expression of caspase-3.5.In nude mice tumorigenesis,nude mice transplanted with SChLAP1 downregulated or miR-101-5p upregulated tumors were smaller and grew more slowly than NC mice.Conclusions:SChLAPl can affect prostate cancer cell proliferation and metastasis by regulating miR-101-5p expression.MiR-101-5p may be one of the potential biomarkers for prostate cancer.