背景:诊断方面的进步,治疗,对去势抵抗前列腺癌(CRPC)的监测已经取得了很大进展,但是结合现有临床和病理数据的新生物标志物可能有助于更精确的诊断和预后。一些研究发现,细胞外囊泡(EV)衍生的miRNA在各种类型的恶性肿瘤中起着至关重要的作用。本研究的目的是探索EVmiRNA并确定其作为CRPC诊断和预后生物标志物的生物学功能。
方法:从五个健康供体收集血浆样本(对照,CT)和17例CRPC患者,根据内分泌治疗反应分为两组:部分反应(PR;n=10)和进行性疾病(PD;n=7)。使用miRNA微阵列和RT-qPCR鉴定候选细胞外囊泡(EV)miRNA。选择的miRNA的生物学功能使用MTT分析进行评估,伤口愈合试验,trans-well分析,和瞬时miRNA表达后CRPC细胞中的RNA测序。
结果:微阵列分析显示,与CT和PR样品相比,PD样品中EV-miR-6880-5p显著下调(p<0.01)。使用RT-qPCR,与CT组相比,CRPC患者中EV-miR-6880-5p的表达降低(p=0.0336)。在PR组中,EV-miR-6880-5p在随访时与基线相比增加(p=0.2803),而在PD组,与基线样本相比,随访时下降(p=0.4356).此外,miR-6880-5p的过表达阻碍了细胞增殖,迁移,和入侵,与肿瘤进展相关的下调途径,同时上调CRPC细胞中与细胞生长和凋亡相关的通路。
结论:EV-miR-6880-5p有望作为CRPC患者的预后生物标志物。Further,前瞻性验证对于评估这些候选miRNA的潜力是必要的。
BACKGROUND: Advancements in the diagnosis, treatment, and surveillance of castration-resistant prostate cancer (CRPC) have progressed considerably, but a new biomarker that combines existing clinical and pathological data could be useful for a more precise diagnosis and prognosis. Some investigations have found that extracellular vesicle (EV)-derived miRNAs play crucial roles in various types of malignant tumors. The objective of this study was to explore EV miRNA and identify its biologic function as a biomarker for the diagnosis and prognosis of CRPC.
METHODS: Plasma samples were collected from five healthy donors (Control, CT) and 17 CRPC patients, categorizing into two groups based on their endocrine treatment response: partial response (PR; n = 10) and progressive disease (PD; n = 7). Candidate extracellular vesicle (EV) miRNAs were identified using miRNA microarray and RT-qPCR. The biological functions of the selected miRNAs were evaluated using the MTT assay, wound healing assay, trans-well assay, and RNA sequencing in CRPC cells after transient miRNA expression.
RESULTS: Microarray analysis revealed a significant downregulation of EV-miR-6880-5p in the PD samples compared to both CT and PR samples (p < 0.01). The expression of EV-miR-6880-5p in CRPC patients was decreased compared with that CT group (p = 0.0336) using RT-qPCR. In the PR group, EV-miR-6880-5p was increased at follow-up compared with the baseline (p = 0.2803), while in the PD group, it decreased at follow-up compared with the baseline samples (p = 0.4356). Furthermore, overexpression of miR-6880-5p hampered cell proliferation, migration, and invasion, downregulated pathways associated with tumor progression, and simultaneously upregulated pathways associated with cell growth and apoptosis in CRPC cells.
CONCLUSIONS: EV-miR-6880-5p shows promise as a prognostic biomarker in patients with CRPC. Further, prospective validations are necessary to evaluate the potential of these candidate miRNAs.