| Background Prostate cancer is the second most common cancer in men worldwide and the fifth leading cause of cancer death.Although the diagnostic methods and treatments for prostate cancer have significantly evolved comparing with the previous decades,intermediate and advanced prostate cancer as well as depression-resistant prostate cancer remain important causes of leading death.Statin is competitive inhibitor of hydroxymethylglutaryl coenzyme A,which inhibits the de novo synthesis of cholesterol.Since their discovery,they have quickly become important drugs for lowering plasma cholesterol level,and especially after their successful synthesis,statins have quickly become the most widely used drugs for lowering cholesterol levels worldwide,with atorvastatin being the most representative.In epidemiological observation,some researchers found that statins can prolong the survival of patients who are suffering from the prostate cancer,but the mechanism of this phenomenon has not been explored clearly.Lnc RNAs are a class of non-coding RNA,which are transcribed by the genome with a length greater than 200 nt.They cannot be used as a template for the translation of proteins.The mechanism of action of lnc RNAs is that they function as a ce RNA to competitively inhibit the interaction between mi RNA and m RNA,and then play the role of regulating m RNA.In tumorigenesis and development,lnc RNAs also play a critical role.Studies related to lnc RNAs,and malignancies can be used as potential targets for tumor therapy,and the basis for prognosis determination,and tumor markers for early diagnosis.ObjectivesIn this study,the effect of atorvastatin on prostate cancer PC-3 cell line was used as an entry point to verify the inhibitory effect of atorvastatin on prostate cancer in vitro and in vivo.To further search for differences in lnc RNA expression in atorvastatin-acting PC-3 cells and to verify its effect.Then to investigate the mechanisms underlying the inhibitory effect of atorvastatin on prostate cancer.Methods and material1)The proliferation inhibitory effect of atorvastatin on PC-3 cells was observed by CCK-8 and clone formation assay.2)Atorvastatin induced apoptosis in PC-3 cells as verified by flow cytometry assay and Western blot for apoptosis key protein content.3)Atorvastatin inhibited migration and invasion of PC-3 cells as verified by wound healing assay and Transwell assay.(4)The key protein levels of E-cadherin,N-cadherin,Vimentin,MMP-2,and MMP-9 were measured by Western blot to verify that atorvastatin inhibited the EMT in PC-3 cells.6)Use high-throughput sequencing to identify lnc RNA differentially expressed in PC-3 cells before and after atorvastatin treatment,then use bioinformatics databases to screen for effective target genes.7)Use si RNA transfection to reduce the expression of target lnc RNA in PC-3 cells,and observe them by CCK-8,clone formation,flow cytometry,and Transwell assays.changes in biological behavior.8)The expression of lnc RNA target genes and their proteins in PC-3 cells was verified by q PCR and Western blot.Results1)CCK-8 and clone formation assays showed that atorvastatin could inhibit the proliferation of PC-3 cells in vitro,and this effect was dose-dependent.2)Flow cytometry and Hoechst 33342 staining results showed that atorvastatin could promote the apoptosis of PC-3 cells.The higher the concentration of atorvastatin,the more significant the pro-apoptotic effect.3)Wound healing assay and Transwell assay showed that atorvastatin could inhibit the migration and invasion ability of PC-3 cells in vitro.4)Atorvastatin can reduce the content of MMP-2 and MMP-9 in PC-3 cells,inhibiting the EMT of PC-3 cells.5)Atorvastatin can inhibit the proliferation and migration of PC-3 cells in vivo as seen in subcutaneous tumorigenesis assay in nude mice.6)High-throughput sequencing screened the differentially expressed lnc RNA of the C-3 cell line before and after the effect of atorvastatin as SNHG4.7)We use the bioinformatics database screening the SNHG4/mi R-206/E2F5 interaction pathway as the target pathway for the following study.8)The expression of mi R-206 is significantly increased after interfering with SNHG4 expression,while the expression of E2F5 was significantly decreased,the E2F5 protein content in PC-3 cells was significantly decreased,and the effect was more significant after adding atorvastatin.9)The proliferation of PC-3 cells was significantly inhibited after the inhibition of SNHG4 expression,and the inhibitory effect was even more significant after the addition of atorvastatin on this basis.10)The apoptosis numbers of PC-3 cells were significantly increased after inhibition of SNHG4 expression,while the apoptosis of tumor cells was more obvious after adding atorvastatin.11)The migration and invasion ability of PC-3 cells was significantly decreased after the inhibition of SNHG4 expression,while the inhibition effect was more significant after adding atorvastatin.Conclusions1.Atorvastatin inhibited the proliferation,migration,and invasive ability of prostate cancer PC-3 cell lines and promoted apoptosis of cancer cells in both in vitro and in vivo assays.2.Atorvastatin affects the EMT of cancer cells and thus the migration and invasion of prostate cancer cells by regulating the expression of MMP-2 and MMP-9.3.Atorvastatin inhibits the expression of SNHG4 in PC-3 cells,and SNHG4 affects the expression of E2F5,the target gene of mi R-206,by competitively inhibiting the expression of mi R-206.4.Atorvastatin can function through the SNHG4/mi R-206/E2F5 interaction axis to inhibit the proliferation,migration,and invasion of PC-3 cells. |
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