CDCA8通过调控增殖促进前列腺癌发生的作用机制研究

Study on the mechanism of CDCA8 promoting the occurrence of prostate cancer by regulating proliferation

目的 探讨细胞分裂周期相关蛋白 8(CDCA8)在前列腺癌(PCa)中的表达及作用机制。方法 通过生物 信息学方法分析正常前列腺组织和 PCa组织中 CDCA8 mRNA水平差异。利用癌症基因组图谱(TCGA)数据库中 RNA表达测序数据分析 CDCA8表达相关的 PCa患者无病生存期(DFS)。免疫组织化学方法检测根治性前列腺切除 术后 56例 PCa组织和癌旁组织中 CDCA8蛋白表达差异,并依据染色指数将 PCa患者分为 CDCA8蛋白高表达组(31 例)和低表达组(25例),分析患者临床特征与 CDCA8表达水平的相关性。利用 siRNA沉默 PCa细胞 CDCA8基因,实 时荧光定量聚合酶链反应(qPCR)检测 CDCA8 mRNA的表达水平,细胞克隆形成实验检测 PCa细胞增殖能力,蛋白 免疫印迹法检测 CDCA8、Ki67、增殖细胞核抗原(PCNA)以及磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶 B(AKT)信号通路 蛋白表达的变化。结果 TCGA数据库中 PCa组织中 CDCA8 mRNA水平明显高于正常组织,CDCA8 mRNA高表达的 患者预后更差(P<0.01)。PCa组织 CDCA8的蛋白表达水平明显高于癌旁组织,且表达水平与患者总前列腺特异性 抗原(tPSA)、Gleason评分、T分期和术后切缘情况呈正相关(P<0.01)。CDCA8蛋白高表达组tPSA≥10 μg/L、T3期、术 后切缘阳性患者比例大于 CDCA8 蛋白低表达组(P<0.05)。沉默 CDCA8 基因后 PCa 细胞增殖能力减弱,Ki67、 PCNA、p-PI3K、p-AKT蛋白表达水平均下调(P<0.01)。结论 CDCA8通过调控增殖促进 PCa发生,其作用机制可 能与 PI3K/AKT信号通路有关。

Objective To explore the expression and mechanism of cell division cycle associated protein 8(CDCA8)in the occurrence of prostate cancer(PCa).Methods The difference of CDCA8 mRNA expression levels between normal prostate tissue and PCa tissue was analyzed by bioinformatics method.The disease-free survival(DFS)associated with CDCA8 expression in PCa patients was analyzed using RNA sequencing data from cancer Genome Atlas(TCGA)database.Immunohistochemical method was used to detect the expression of CDCA8 in 56 pairs of PCa tissues and adjacent tissues after radical prostatectomy.Patients were divided into the CDCA8 high expression group(n=31)and the CDCA8 low expression group(n=25)according to staining results.The potential correlation between clinical characteristics and CDCA8 expression level was further analyzed.The CDCA8 gene was silenced by siRNA,and the expression level of CDCA8 mRNA was detected by qPCR.The proliferation of PCa cells was detected by clonal formation assay,and the changes of CDCA8,Ki67,proliferating cell nuclear antigen(PCNA)and phosphatidylinositol-3-kinase(PI3K)/protein kinase B(AKT)signaling pathway proteins were detected by Western blot assay.Results In TCGA database,CDCA8 mRNA level was significantly higher in PCa tissues than that in normal tissues,and patients with high expression of CDCA8 mRNA had a worse prognosis(P<0.01).The staining index of expression level of CDCA8 protein was significantly higher in PCa tissue than that in para cancer tissue,and the expression level was positively correlated with total prostate-specific antigen(tPSA),Gleason score,T stage and surgical margin(P<0.01).The proportion of patients with tPSA≥10μg/L,T3 stage and positive postoperative resection margin was significantly higher in the high CDCA8 expression group than that in the low CDCA8 expression group(P<0.05).After CDCA8 gene was silenced,the proliferation of PCa cells was decreased,and the protein expression levels of Ki67,PCNA,p-PI3K and p-AKT were down-regulated(P<0.01).Conclusion CDCA8 promotes the occurrence of prostate cancer by regulating proliferation,and its mechanism may be related to PI3K/AKT signaling pathway.