LncRNA SNHG7 Promotes Prostate Cancer Progression Via Targeting MiR-503

PurposesProstate cancer is a common malignancy in men in many countries of the world.Millions of people around the world suffer from this disease.The incidence of prostate cancer is closely related to age,and the incidence of elderly men is the highest.1 5%of patients worldwide have a family history of prostate cancer.Prostate cancer is the second leading cause of death among males in the United States.In recent years,the incidence of prostate cancer in China has also increased rapidly.According to the cancer detection data published in 2015,the incidence of prostate cancer in China ranks 7th among male cancers.and the mortality rate ranks 12th among male cancers.In the pathological type of prostate cancer,the incidence of adenocarcinoma accounts for more than half.In order to elucidate the regulation mechanism of lncRNA related to prostate cancer and to find potential clinical signs and potential therapeutic targets for prostate cancer,our research group obtained lncRNASNHG7 through TCGA and Taylor prostate cancer patient database data analysis and gene cluster enrichment analysis.And miR-503 is associated with the development of prostate cancer.As a result,it was found that the expression level of IncRNA SNHG7 and in prostate cancer tissues was increased,and the expression level of miR-503 was decreased in prostate cancer tissues.This study firstly confirmed the expression of lncRNA SNHG7 and miR-503 in prostate cancer and the effect of lncRNA SNHG7 on prostate cancer cell proliferation,cell cycle,apoptosis and cell phenotype.And then we further studied the interaction of lncRNA SNHG7 with miR-503 and the effect of IncRNA SNHG7 on miR-503 target gene.Finally,we explored the molecular mechanism of lncRNA SNHG7 affecting the phenotypic changes of prostate cancer cells.Studying the pathogenesis of prostate cancer can provide sensitive and accurate tumor markers for early clinical diagnosis,and provide effective therapeutic targets for the clinic.Methods1.The expression of IncRNA SNHG7 and miR-503 in clinical prostate cancer tissues of different stages(Phase ?,?,?)was detected by qRT-PCR,and it was verified whether IncRNA SNHG7 and miR-503 were associated with the development of prostate cancer.2.The proliferation,apoptosis and other cell phenotypes of prostate cancer cells from three different sources were compared by the following experimental methods:cell viability was detected by CCK8 cell proliferation assay;cell clone formation rate was detected by plate clone assay;cell invasion ability was detected by Transwell assay;The trace(ibidi plug-in)assay was used to detect cell migration ability;Annexin V-FITC/PI flow double staining was used to detect apoptosis rate;qRT-PCR was used to detect the expression of IncRNA SNHG7 and miR-503.3.Overexpression/interference of IncRNA SNHG7 in PC-3 cells,cell clone formation rate was detected by plate cloning assay;apoptosis was detected by Annexin V-FITC/PI double staining flow cytometry;PI single staining was used to detect cell cycle;K167 and DAPI immunofluorescence double staining method for cell proliferation;tumor cell angiogenesis assay;qRT-PCR detection of IncRNA SNHG7 and miR-503 expression.To investigate the effect of IncRNA SNHG7 on proliferation,cell cycle,apoptosis and cell phenotype of prostate cancer cell line PC-3 in vitro.4.We used IncRNA SNHG7 overexpression and interference with lentivirus to infect PC-3 cells,and used puromycin to screen stably expressed cell lines.A stable cell strain was transplanted subcutaneously into nude mice to establish an animal model of subcutaneous tumor formation.The effect of IncRNA SNHG7 on tumor proliferation was investigated by measuring the volume and weight of tumor tissue and the expression and distribution of ki67 by immunohistochemistry.5.Through RIP experiments to study the possible interaction of IncRNA SNHG7,miR-503,further study the interaction of IncRNA SNHG7 and miR-503 by dual luciferase assay.According to the literature,PI3K is a target protein of miR-503,and we further verified this conclusion by double luciferase assay.In order to verify that IncRNA SNHG7 affects the proliferation,apoptosis and cell phenotype of prostate cancer by inhibiting the target gene miR-503,we restored the expression of miR-503 in prostate cancer cell line PC-3,respectively,by plate cloning.The cell clone formation rate was detected by the experiment;apoptosis was detected by flow cytometry by Annexin V-FITC/PI double staining;cell cycle was detected by PI single staining;cell proliferation was detected by ki67 and DAPI immunofluorescence double staining;cell scratches(ibidi plug-in)Experimental detection of cell migration ability;transwell assay to detect cell invasion ability;determination of tumor cell angiogenesis experiments.In order to study the molecular mechanism of IncRNA affecting the phenotypic changes of prostate cancer cells,we changed the expression levels of IncRNA SNHG7 and miR-503,and detected the PI3-K/Akt pathway through WB with cell proliferation,apoptosis,migration and invasion-related proteins,expression.Results1.The expression of lncRNA SNHG7 was up-regulated in prostate cancer tissues,with the most up-regulated phase ? being the most significant,followed by phase ?;miR-503 was down-regulated in prostate cancer tissues,with phase ? being the most significant,followed by phase ?..2.LncRNA SNHG7 has the highest expression in PC-3 cells,and miR-503 has the lowest expression in PC-3 cells.At the same time,PC-3 cells had the strongest proliferation,migration and invasion ability,and the lowest level of apoptosis.3.In vitro,up-regulation of IncRNA SNHG7 promotes proliferation,migration and invasion of prostate cancer cell line PC-3,and inhibits apoptosis.4.1n vivo,up-regulation of lncRNA SNHG7 promotes prostate cancer tumor proliferation and inhibits its apoptosis.5.LncRNA SNHG7 and miR-503 bind to the AG02 protein to form a ternary complex.lncRNA SNHG7 regulates the expression of its target gene PI3K by targeting inhibition of miR-503.6.The effect of IncRNA SNHG7 on tumor cell proliferation,apoptosis and cell phenotype was achieved by inhibiting miR-503.1.LncRNA SNHG7 targets the inhibition of miR-503 and thus affects tumor cell proliferation,apoptosis and cell phenotype by activating PI3K/AKT-related pathways.Conclusions1.Abnormal expression of lncRNA SNHG7 and miR-503 in prostate cancer is associated with the development of prostate cancer.The more severe the prostate cancer,the higher the expression of IncRNA SNHG7 and the lower the expression of miR-503.2.LncRNA SNHG7 promotes prostate cancer proliferation,migration and invasion,inhibition of apoptosis.3.LncRNA SNHG7 regulates the expression of its target gene PI3K by targeting inhibition of miR-503.The effect of IncRNA SNHG7 on tumor cell proliferation.apoptosis and cell phenotype was achieved by inhibiting miR-503.4.LncRNA SNHG7 targeting inhibition of miR-503 and activation of PI3K/AKT signaling pathway affect tumor cell proliferation,apoptosis and cell phenotype.