目的?探讨肾透明细胞癌(clear cell renal cell carcinoma，ccRCC)患者血清细胞外囊泡(extracellular vesicles，EVs)中miR-99b-3p的水平变化，评估其作为ccRCC非侵入性辅助诊断分子标志物的临床价值。方法?收集2016年12月至2018年10月东部战区总医院泌尿外科收治且未经治疗的65例ccRCC患者，另收集75例年龄、性别匹配的健康人对照血清标本，分离血清EVs并提取RNA。采用实时荧光定量PCR(qRT-PCR)检测ccRCC患者及健康人对照血清EVs miR-99b-3p的水平，并采用Mann-Whitney?U检验、ROC曲线及Logistic回归分析EVs miR-99b-3p水平对ccRCC的诊断效能，Pearson相关分析EVs miR-99b-3p水平与ccRCC患者临床病理参数的相关性。结果?Mann-Whitney?U检验结果显示，ccRCC患者血清EVs miR-99b-3p的表达水平较健康人对照组明显上调(P<0.01)；ROC曲线分析显示，EVs miR-99b-3p筛查ccRCC患者与健康人对照的曲线下面积(AUCROC)为0.757(95%CI：0.674～0.840)，敏感性为69.2%，特异性为77.3%；区分Ⅰ/Ⅱ级ccRCC患者的AUCROC为0.754(95%CI：0.663～0.846)，敏感性和特异性分别为69.2%、77.3%；Logistic回归分析结果显示，血清EVs miR-99b-3p是ccRCC独立危险因素(OR=7.745，95%CI：3.596～16.682，P<0.01)；Pearson相关分析显示，血清EVs miR-99b-3p与肿瘤分级、大小、有无远处转移的相关性均无统计学意义(P均>0.05)。结论?ccRCC患者血清EVs miR-99b-3p水平较健康人对照显著升高，是ccRCC潜在的液体活检辅助诊断分子标志物。
Objective?To investigate the expression of miR-99b-3p in serum extracellular vesicles (EVs) of the patients with clear cell renal cell carcinoma (ccRCC) and evaluate its clinical value as a noninvasive diagnostic marker for ccRCC.?Methods?A total of 65 ccRCC patients from Department of Urology and 75 age-sex matched healthy individuals from Health Examination Center of General Hospital of Eastern Theater Command from December 2016 to October 2018 were enrolled in this study. Serum EVs was isolated from the patients and healthy individuals, and the level of EVs miR-99b-3p was determined by qRT-PCR. The clinical value of serum EVs miR-99b-3p was further assessed by Mann-Whitney?U?test, receiver operating characteristic curve (ROC) and logistics analyses. Pearson correlation analysis method was used to analyze the correlation between EVs miR-99b-3p level and ccRCC clinicopathological parameters.?Results?According to the Mann-Whitney?U?test, the relative expression level of serum EVs miR-99b-3p in the ccRCC patients was significantly higher than that in the healthy individuals (P<0.01). The level of receive operating characteristic curve (ROC) analysis demonstrated that the efficiency of area under curve (AUCROC) for discriminating the ccRCC patients from healthy individuals was 0.757 (95%CI: 0.674 to 0.840) with sensitivity of 69.2% and specificity of 77.3% at the optimal cutoff value. The AUCROC?for differentiating early stage ccRCC patients from healthy individuals was 0.754 (95%CI: 0.663 to 0.846) with sensitivity of 69.2% and specificity of 77.3% at the same cutoff value. Logistic regression analyses showed that serum EVs miR-99b-3p was an independent risk factor for ccRCC (OR=7.745, 95%CI: 3.596 to 16.682,?P<0.01). Pearson correlation analysis revealed that no statistically significant correlations between serum EVs miR-99b-3p and the stage and size of tumor as well as the distant metastasis (P>0.05).?Conclusion?Serum EVs miR-99b-3p levels may significantly increased in ccRCC patients, and could be served as a novel non-invasive liquid biopsy diagnostic marker for ccRCC.