Breyer Johannes, Wirtz Ralph M, Otto Wolfgang, Erben Philipp, Kriegmair Maximilian C, Stoehr Robert, Eckstein Markus, Eidt Sebastian, Denzinger Stefan, Burger Maximilian, Hartmann Arndt
Department of Urology, University of Regensburg, Caritas St. Josef Medical Center Landshuter Str. 65, 93053, Regensburg, Germany.
STRATIFYER Molecular Pathology GmbH, Cologne, Germany.
Virchows Arch. 2017 Mar;470(3):267-274. doi: 10.1007/s00428-017-2064-8. Epub 2017 Jan 10.
Differential expression of cytokeratins (CK) is a characteristic feature of chemoresistant luminal (KRT20) and chemosensitive intrinsic aggressive basal (KRT5) subtypes in muscle-invasive bladder cancer (MIBC). We investigated mRNA expression of KRT5 and KRT20 and its predictive value in stage pT1 bladder cancer. In retrospective analysis of clinical data and formalin-fixed paraffin-embedded tissues (FFPE) of patients with stage pT1 NMIBC who underwent transurethral resection of the bladder, a single-step RT-qPCR was used to measure mRNA expression. Furthermore, immunohistochemical (IHC) staining of CK20, panCK, and MIB1 was performed. Valid measurements were obtained from 231 samples out of a series of 284 patients. Spearman correlation revealed significant associations between mRNA and protein expression of KRT20/CK20 (ρ 0.6096, p < 0.0001) and MKI67/MIB1 (ρ 0.5467, p < 0.0001). A positive correlation was found between MKI67 and KRT20 expression (ρ 0.3492, p < 0.0001), while MKI67 and KRT5 were negatively correlated (ρ -0.1693, p = 0.01). High KRT20 expression (≥40.26) was significantly associated with worse recurrence free survival (RFS) (p = 0.001), progression-free survival (PFS) (p = 0.0003), and cancer specific survival (CSS) (p = 0.0414). The combination of high KRT20 expression and low KRT5 expression (<36.83) was associated with unfavorable RFS (p = 0.0038) and PFS (p = 0.0003) and proved to be the only independent predictor for RFS (p = 0.0055) and PFS (p = 0.0023) in multivariate analysis. KRT20 mRNA determination was superior to CK20 protein estimation with regard to RFS and PFS prediction. KRT20 and KRT5 mRNA quantification can predict recurrence and progression of stage pT1 NMIBC reflecting basal and luminal subtypes of MIBC and is superior to CK20 protein expression determined by IHC.
细胞角蛋白（CK）的差异表达是肌肉浸润性膀胱癌（MIBC）中化疗耐药的管腔型（KRT20）和化疗敏感的侵袭性基底型（KRT5）亚型的一个特征。我们研究了KRT5和KRT20的mRNA表达及其在pT1期膀胱癌中的预测价值。在对接受经尿道膀胱切除术的pT1期非肌层浸润性膀胱癌（NMIBC）患者的临床资料和福尔马林固定石蜡包埋组织（FFPE）进行回顾性分析时，采用单步RT-qPCR来测量mRNA表达。此外，还进行了CK20、泛CK和MIB1的免疫组织化学（IHC）染色。在一系列284例患者中，从231个样本中获得了有效的测量结果。Spearman相关性分析显示KRT20/CK20的mRNA和蛋白表达之间存在显著关联（ρ = 0.6096，p < 0.0001）以及MKI67/MIB1之间存在显著关联（ρ = 0.5467，p < 0.0001）。发现MKI67与KRT20表达呈正相关（ρ = 0.3492，p < 0.0001），而MKI67与KRT5呈负相关（ρ = -0.1693，p = 0.01）。高KRT20表达（≥40.26）与无复发生存期（RFS）较差（p = 0.001）、无进展生存期（PFS）较差（p = 0.0003）以及癌症特异性生存期（CSS）较差（p = 0.0414）显著相关。高KRT20表达和低KRT5表达（<36.83）的组合与不良的RFS（p = 0.0038）和PFS（p = 0.0003）相关，并且在多变量分析中被证明是RFS（p = 0.0055）和PFS（p = 0.0023）的唯一独立预测因子。在RFS和PFS预测方面，KRT20 mRNA测定优于CK20蛋白评估。KRT20和KRT5 mRNA定量可以预测pT1期NMIBC的复发和进展，反映MIBC的基底型和管腔型亚型，并且优于通过IHC测定的CK20蛋白表达。
