Department of Cell Biology and Biophysics, Faculty of Biology, National and Kapodistrian University of Athens, 15784 Athens, Greece.
Department of Biochemistry and Molecular Biology, Faculty of Biology, National and Kapodistrian University of Athens, 15701 Athens, Greece.
Int J Mol Sci. 2023 Jan 30;24(3):2609. doi: 10.3390/ijms24032609.
Bladder cancer (BlCa) represents the sixth most commonly diagnosed type of male malignancy. Due to the clinical heterogeneity of BlCa, novel markers would optimize treatment efficacy and improve prognosis. The small heat shock proteins (sHSP) family is one of the major groups of molecular chaperones responsible for the maintenance of proteome functionality and stability. However, the role of sHSPs in BlCa remains largely unknown. The present study aimed to examine the association between and expression and BlCa progression in patients, and to investigate their role in BlCa cells. For this purpose, a series of experiments including reverse transcription-quantitative PCR, Western blotting, MTT assay and flow cytometry were performed. Initial analyses revealed increased vs. human transitional carcinoma cells, expression levels of the and genes and proteins in high grade BlCa cell lines. Therefore, we then evaluated the clinical significance of the and genes expression levels in bladder tumor samples and matched adjusted normal bladder specimens. Total RNA from 100 bladder tumor samples and 49 paired non-cancerous bladder specimens were isolated, and an accurate SYBR-Green based real-time quantitative polymerase chain reaction (qPCR) protocol was developed to quantify and mRNA levels in the two cohorts of specimens. A significant downregulation of the and genes expression was observed in bladder tumors as compared to matched normal urothelium; yet, increased and levels were noted in muscle-invasive (T2-T4) vs. superficial tumors (TaT1), as well as in high-grade vs. low-grade tumors. Survival analyses highlighted the significantly higher risk for post-treatment disease relapse in TaT1 patients poorly expressing and genes; this effect tended to be inverted in advanced disease stages (muscle-invasive tumors) indicating the biphasic impact of , genes in BlCa progression. The pro-survival role of and in advanced tumor cells was also evident by our finding that , genes expression silencing in high grade BlCa cells enhanced doxorubicin toxicity. These findings indicate that the , chaperone genes have a likely pro-survival role in advanced BlCa; thus, they can be targeted as novel molecular markers to optimize treatment efficacy in BlCa and to limit unnecessary interventions.
膀胱癌（BlCa）是男性最常见的第六种恶性肿瘤。由于 BlCa 的临床异质性，新的标志物将优化治疗效果并改善预后。小分子热休克蛋白（sHSP）家族是负责维持蛋白质组功能和稳定性的主要分子伴侣之一。然而，sHSPs 在 BlCa 中的作用在很大程度上仍是未知的。本研究旨在探讨 和 在患者 BlCa 进展中的表达与 BlCa 进展之间的关系，并探讨它们在 BlCa 细胞中的作用。为此，进行了一系列实验，包括逆转录-定量 PCR、Western blot、MTT 测定和流式细胞术。初步分析显示，在高级别 BlCa 细胞系中， 和 基因和蛋白质的表达水平升高。因此，我们评估了 和 基因在膀胱肿瘤样本和配对的调整正常膀胱标本中的表达水平的临床意义。从 100 例膀胱肿瘤样本和 49 例配对的非癌性膀胱标本中分离总 RNA，并开发了一种准确的 SYBR-Green 基于实时定量聚合酶链反应（qPCR）方案，以定量两个队列标本中的 和 mRNA 水平。与匹配的正常尿路上皮相比，在膀胱肿瘤中观察到 和 基因表达显著下调；然而，在肌肉浸润性（T2-T4）与浅表性肿瘤（TaT1）以及高级别与低级别肿瘤中， 和 水平增加。生存分析突出表明，在 TaT1 患者中， 和 基因表达水平低的患者在治疗后疾病复发的风险明显更高；这种效应在晚期疾病阶段（肌肉浸润性肿瘤）中趋于反转，表明 ，在 BlCa 进展中， 基因具有双相影响。在高级别 BlCa 细胞中， 和 基因表达沉默增强阿霉素毒性的发现也表明了 和 基因在高级别肿瘤细胞中的促生存作用。这些发现表明， ， 伴侣基因在晚期 BlCa 中可能具有促生存作用；因此，它们可以作为新的分子标志物，以优化 BlCa 的治疗效果，并限制不必要的干预。
