抑制CDKN2B提高前列腺癌细胞对恩杂鲁胺的化学敏感性

An Evaluation of the Suppression of CDKN2B Contributes to the Recovery ofChemosensitivity of Enzalutamide-resistant Prostate Cancer Cells

目的探究前列腺癌(prostate cancer,PCa)患者对恩杂鲁胺(Enzalutamide,Enz)化疗抵抗的机制。方法从GEO数据库中选定GSE78201表达谱,利用生物信息学筛选出导致恩杂鲁胺抵抗的候选基因。利用PCa细胞系C4-2建立恩杂鲁胺抵抗细胞系Enz-R。使用shRNA和反义寡核苷酸(ASO)在Enz-R细胞系中对CDKN2B进行敲低,通过克隆形成、MTT和细胞毒性实验检测抑制CDKN2B后Enz-R细胞对恩杂鲁胺的抗性。动物实验检测敲低CDKN2B后,恩杂鲁胺对前列腺肿瘤的治疗效果。结果CDKN2B在候选基因中表达上调最为显著,且它的上调与PCa患者的不良预后相关。敲低CDKN2B的Enz-R细胞经恩杂鲁胺处理后集落形成数目减少,生长速率减慢,对恩杂鲁胺的耐药性减弱。体内实验证明,敲低CDKN2B并使用恩杂鲁胺进行治疗,可显著抑制PCa肿瘤的生长。结论抑制CDKN2B可提高PCa细胞对恩杂鲁胺的敏感性,CDKN2B可作为PCa的潜在治疗靶点。

Objective The aim of this study was to investigate the underlying mechanism of resistance to enzalutamide(Enz)chemotherapy in patients with prostate cancer(PCa).Methods In this study,we selected GSE78201 expression profiles from GEO database and used bioinformatics approach to screen for candidate genes that could cause Enz resistance.We developed enzalutamide-resistant cell line Enz-R using PCa cell line C4-2.CDKN2B was knocked down in Enz-R cell lines using shRNA and antisense oligonucleotides(ASO),and Enz-R cells were tested for the resistance to Enz after inhibition of CDKN2B by clone formation,MTT and cytotoxicity assays.Animal assays were performed to detect the therapeutic effect of Enz on prostate tumors after knockdown of CDKN2B.Results CDKN2B expression was most significantly upregulated among the candidate genes and its upregulation was associated with the poor prognosis in PCa patients.Enz-R cells with knockdown CDKN2B showed reduced number of colony formation,slower growth rate and weakened resistance to Enz after Enz treatment.In vivo experiments demonstrated that knockdown of CDKN2B and treatment with Enz significantly inhibited the growth of PCa tumors.Conclusion Inhibition of CDKN2B could improve the sensitivity of PCa cells to Enz,and CDKN2B could serve as a potential therapeutic target for PCa.