Division of Urology, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei 11490, Taiwan.
Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei 11490, Taiwan.
Int J Mol Sci. 2022 Feb 13;23(4):2072. doi: 10.3390/ijms23042072.
Urothelial carcinoma includes upper urinary tract cancer (UTUC) and bladder cancer. Although nephroureterectomy is the standard treatment for UTUC, the recurrence rate is approximately half and the tumor is associated with poor prognoses. Metastases are the most devastating and lethal clinical situation in urothelial carcinoma. Despite its clinical importance, few potential diagnostic biomarkers are suitable for early UC detection. We compared high-stage/high-grade urothelial carcinoma tissues to adjacent normal urothelial tissues using methyl-CpG binding domain protein capture for genome-wide DNA methylation analysis. Based on our findings, inhibin βA (INHBA) might be associated with carcinogenesis and metastasis. Further, clinical UC specimens had significant INHBA hypomethylation based on pyrosequencing. INHBA was detected by real-time PCR and immunohistochemistry staining, and was found to be highly expressed in clinical tissues and cell lines of urothelial carcinoma. Further, INHBA depletion was found to significantly reduce BFTC-909 cell growth and migration by INHBA-specific small interfering RNA. Interestingly, a positive correlation was found between SMAD binding and extracellular structure organization with INHBA using gene set enrichment analysis and gene ontology analysis. Together, these results are the first evidence of INHBA promoter hypomethylation and INHBA overexpression in UTUC. INHBA may affect urothelial carcinoma migration by reorganizing the extracellular matrix through the SMAD pathway.
尿路上皮癌包括上尿路尿路上皮癌（UTUC）和膀胱癌。虽然肾输尿管切除术是 UTUC 的标准治疗方法，但复发率约为一半，且肿瘤预后较差。转移是尿路上皮癌最具破坏性和致命的临床情况。尽管其具有重要的临床意义，但很少有潜在的诊断生物标志物适合早期 UC 检测。我们使用甲基化CpG 结合域蛋白捕获进行全基因组 DNA 甲基化分析，比较了高分期/高级别尿路上皮癌组织与相邻正常尿路上皮组织。根据我们的发现，抑制素βA（INHBA）可能与癌发生和转移有关。此外，基于焦磷酸测序，临床 UC 标本存在显著的 INHBA 低甲基化。通过实时 PCR 和免疫组织化学染色检测到 INHBA，并在临床尿路上皮癌组织和细胞系中发现其高表达。此外，通过 INHBA 特异性小干扰 RNA 发现 INHBA 耗竭可显著降低 BFTC-909 细胞的生长和迁移。有趣的是，使用基因集富集分析和基因本体论分析发现，INHBA 与 SMAD 结合和细胞外结构组织之间存在正相关。总之，这些结果首次证明了 UTUC 中 INHBA 启动子的低甲基化和 INHBA 的过表达。INHBA 可能通过 SMAD 途径重组细胞外基质来影响尿路上皮癌的迁移。
