Koukourikis Periklis, Papaioannou Maria, Georgopoulos Petros, Apostolidis Ioannis, Pervana Stavroula, Apostolidis Apostolos
2nd Department of Urology, Aristotle University of Thessaloniki, General Hospital 'Papageorgiou', 56403 Thessaloniki, Greece.
Department of Biological Chemistry, Medical School, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Biology (Basel). 2023 Aug 12;12(8):1126. doi: 10.3390/biology12081126.
Bladder cancer (BCa) in patients suffering from neurogenic lower urinary tract dysfunction (NLUTD) is a significant concern due to its advanced stage at diagnosis and high mortality rate. Currently, there is a scarcity of specific guidelines for BCa screening in these patients. The development of urine biomarkers for BCa seems to be an attractive non-invasive method of screening or risk stratification in this patient population. DNA methylation is an epigenetic modification, resulting in the transcriptional silencing of tumor suppression genes, that is frequently detected in the urine of BCa patients. We aimed to investigate DNA hypermethylation in five gene promoters, previously associated with BCa, in the urine of NLUTD patients, and in comparison with healthy controls. This was a prospective case-control study that recruited neurourology outpatients from a public teaching hospital who had suffered from NLUTD for at least 5 years. They all underwent cystoscopy combined with biopsy for BCa screening following written informed consent. DNA was extracted and DNA methylation was assessed for the RASSF1, RARβ, DAPK, TERT and APC gene promoters via quantitative methylation-specific PCR in urine specimens from the patients and controls. Forty-one patients of mixed NLUTD etiology and 35 controls were enrolled. DNA was detected in 36 patients' urine specimens and in those of 22 controls. In the urine specimens, DNA was hypermethylated in at least one of five gene promoters in 17/36 patients and in 3/22 controls (47.22% vs. 13.64%, respectively, = 0.009). RASSF1 was hypermethylated in 10/17 (58.82%) specimens with detected methylation, APC in 7/17 (41.18%), DAPK in 4/17 (23.53%), RAR-β2 in 3/17 (17.56%) and TERT in none. According to a multivariate logistic regression analysis, NLUTD and male gender were significantly associated with hypermethylation (OR = 7.43, = 0.007 and OR = 4.21; = 0.04, respectively). In the tissue specimens, histology revealed TaLG BCa in two patients and urothelial squamous metaplasia in five patients. Chronic bladder inflammation was present in 35/41 bladder biopsies. DNA hypermethylation in a panel of five BCa-associated genes in the urine was significantly more frequent in NLUTD patients than in the controls. Our results warrant further evaluation in longitudinal studies assessing the clinical implications and possible associations between DNA hypermethylation, chronic inflammation and BCa in the NLUTD population.
神经源性下尿路功能障碍（NLUTD）患者的膀胱癌（BCa）因其诊断时处于晚期且死亡率高而备受关注。目前，针对这些患者的BCa筛查缺乏具体指南。开发用于BCa的尿液生物标志物似乎是该患者群体中一种有吸引力的非侵入性筛查或风险分层方法。DNA甲基化是一种表观遗传修饰，会导致肿瘤抑制基因转录沉默，在BCa患者尿液中经常被检测到。我们旨在研究NLUTD患者尿液中先前与BCa相关的五个基因启动子中的DNA高甲基化情况，并与健康对照进行比较。这是一项前瞻性病例对照研究，招募了一家公立教学医院的神经泌尿外科门诊患者，这些患者患有NLUTD至少5年。在获得书面知情同意后，他们均接受了膀胱镜检查并结合活检进行BCa筛查。从患者和对照的尿液标本中提取DNA，并通过定量甲基化特异性PCR评估RASSF1、RARβ、DAPK、TERT和APC基因启动子的DNA甲基化情况。纳入了41例病因混合的NLUTD患者和35例对照。在36例患者的尿液标本和22例对照的尿液标本中检测到了DNA。在尿液标本中，17/36例患者的五个基因启动子中至少有一个发生了DNA高甲基化，3/22例对照中也有这种情况（分别为47.22%和13.64%，P = 0.009）。在检测到甲基化的17个标本中，10/17（58.82%）的RASSF1发生了高甲基化，7/17（41.18%）的APC发生了高甲基化，4/17（23.53%） 的DAPK发生了高甲基化，3/17（17.56%）的RAR-β2发生了高甲基化，TERT未发生高甲基化。根据多因素逻辑回归分析，NLUTD和男性性别与高甲基化显著相关（OR = 7.43，P = 0.007和OR = 4.21；P = 0.04）。在组织标本中，组织学检查显示2例患者为TaLG BCa，5例患者为尿路上皮鳞状化生。41例膀胱活检中有35例存在慢性膀胱炎症。NLUTD患者尿液中一组五个与BCa相关基因的DNA高甲基化明显比对照组更常见。我们的结果值得在纵向研究中进一步评估，以评估DNA高甲基化、慢性炎症和NLUTD人群中BCa之间的临床意义和可能的关联。
