miR-129用于前列腺癌诊治及预测临床预后的研究

miR-129 in the diagnosis,treatment and prediction of clinical prognosis of prostate cancer

目的:通过观察miR-129与前列腺癌(PCa)进展的相关关系,探究miR-129在PCa诊治及预测临床预后中的价值。方法:回顾性分析2014年1月至2022年1月于东部战区总医院泌尿外科就诊的310例男性患者。其中健康对照组80例,BPH组80例,PCa组150例(均接受根治性前列腺切除术,未接受化疗、放疗或雄激素剥夺治疗)。留取受试者的血液标本,采用实时荧光定量PCR检测310名受试者的血清miR-129表达水平并分析其有无统计学差异。收集150例PCa患者的原发性PCa组织和对应的癌旁前列腺组织(距离病灶≥3 cm)样本,所有PCa组织的病理学分级均由经验丰富的病理学家完成。记录每位患者的人口学数据和临床参数,包括年龄、体质指数(BMI)、临床分期、Gleason评分、手术边缘状态、血管淋巴浸润状态、精囊浸润状态和生化复发(BCR)情况。采用RT-qPCR测定组织样本中的miR-129含量,分析其与上述人口学数据和临床参数的关系。选用PC-3和DU-145细胞系并分别转染miR-129 precursor或miR-129 scramble。通过细胞增殖测定和蛋白质印迹分析评估各组细胞系的增殖能力及相关蛋白的表达水平,比较分析其结果。结果:与健康男性和BPH患者相比,PCa患者的血清miR-129表达水平显著降低(P<0.01);与癌旁前列腺组织相比,PCa组织中miR-129表达水平显著降低(P均<0.01)。miR-129的表达水平与术前血清特异性抗原(PSA)水平(P<0.001)、组织学分级(P<0.001)、病理分期(P<0.001)、Gleason评分(P<0.001)、淋巴结转移(P=0.02)、血管淋巴侵袭(P=0.021)和生化复发(P=0.001)均呈负相关。Kaplan-Meier分析表明,PCa组织中低miR-129表达水平与较低的无BCR生存率密切相关。多变量生存分析表明,miR-129表达水平可能是PCa患者无BCR生存率的独立预后指标(P<0.001)。高水平的miR-129可能通过调控细胞周期调节蛋白的表达显著抑制PCa细胞的生长。结论:miR-129水平在PCa患者的血清及PCa组织中均显著下调。miR-129水平的降低在PCa细胞的增殖、肿瘤进展中有重要作用,并与低无BCR生存率密切相关。因此,miR-129可被认为是一种新的PCa诊治及预测临床预后的生物标志物。

Objective:To explore the value of miR-129 in the diagnosis,treatment and prediction of the clinical prognosis of PCa by observing the correlation between miR-129 and the progression of the malignancy.Methods:This retrospective analysis included 310 male patients who visited the Department of Urology of the General Hospital of Eastern Theater Command from January 2014 to January 2022,80 as normal healthy men,80 with BPH,and the other 150 with PCa treated by radical prostatectomy without chemotherapy,radiotherapy or androgen-deprivation therapy.We determined the miR-129 expression in the serum and prostatic tissue of all the subjects by real-time quantitative PCR(RT-qPCR),performed pathological grading of the primary cancerous and pericancerous(≥3 cm from the focus)prostate tissues from the 150 PCa patients,collected the demographic and clinical data on all the subjects,and analyzed the correlation of their demographic data with the clinical parameters.We selected and transfected PC-3 and DU-145 cell lines with miR-129 precursor or miR-129 scramble,assessed the proliferation ability of each cell line and detected the expression levels of related proteins by cell proliferation assay and Western blot.Results:The expression of miR-129 was significantly decreased in the serum of the PCa patients compared with that in the normal healthy men and BPH patients(P<0.01),so was it in the PCa tissue in comparison with that in the pericancerous prostatic tissue(P<0.01),and it was negatively correlated with the preoperative serum PSA level(P<0.001),histological grade(P<0.001),pathological stage(P<0.001),Gleason score(P<0.001),lymph node metastasis(P=0.02),angiolymphatic invasion(P=0.021)and biochemical recurrence(BCR)(P=0.001).Kaplan-Meier analysis showed a strong correlation of down-regulated miR-129 expression with a lower BCR-free survival rate,while multivariate survival analysis indicated that the expression of miR-129 was an independent prognostic indicator of BCR-free survival of the PCa patients(P<0.001).Highly expressed miR-129 significantly inhibited the proliferation of PCa cells by regulating the expressions of cell cycle-regulatory proteins.Conclusion:The expression of miR-129 is significantly down-regulated in PCa tissues,which plays an important role in inhibiting the proliferation of PCa cells and tumor progression and improving BCR-free survival.Therefore,miR-129 can be considered as a new independent biomarker for the diagnosis,treatment and prediction of the clinical prognosis of PCa.