Lindner Andrea Katharina, Furlan Tobias, Orme Jacob J, Tulchiner Gennadi, Staudacher Nina, D'Andrea David, Culig Zoran, Pichler Renate
Department of Urology, Medical University of Innsbruck, Anichstrasse 35, 6020 Innsbruck, Austria.
Division of Medical Oncology, Mayo Clinic, First Street SW, Rochester, MN 55905, USA.
J Clin Med. 2022 Apr 15;11(8):2208. doi: 10.3390/jcm11082208.
Platinum-based chemotherapy is the standard of care with concern to first-line systemic therapy for metastatic disease in urothelial cancer (UC). Resistance to chemotherapy despite an initial response is linked with the ability to remove platinum-based DNA adducts and to repair chemotherapy-induced DNA lesions by various DNA repair proteins. The Rad9-Rad1-HUS1 complex that is loaded onto DNA at sites of damage is involved in checkpoint activation as well as DNA repair. Here, we addressed for the first time the potential influence of HUS1 expression in urothelial carcinogenesis (using two human basal urothelial cancer cell lines UM-UC-3 and HT1197) and its role as a potential therapeutic target for predicting responses to platinum-based chemotherapy. Specific inhibition of HUS1 expression in both cell lines was achieved by specific siRNA and validated by Western blot. In order to define the possible importance of HUS1 in the regulation of cellular proliferation, parental and resistant cells were treated with increasing concentrations of either control or HUS1 siRNA. HUS1 protein expression was observed in both human basal urothelial cancer cell lines UM-UC-3 and HT1197. In cisplatin-sensitive cells, knock-down of HUS1 inhibited cellular proliferation in the presence of cisplatin. On the contrary, knock-down of HUS1 in resistant cells did not result in a re-sensitization to cisplatin. Finally, RNAseq data from the Cancer Genome Atlas provided evidence that HUS1 expression is a significant prognostic factor for poor survival in UC patients. In summary, HUS1 may acts as an oncogene in UC and might be a key determinant of the cellular response to cisplatin-based chemotherapy.
对于尿路上皮癌(UC)转移性疾病的一线全身治疗,铂类化疗是标准的治疗方法。尽管最初有反应,但对化疗的耐药性与去除铂类DNA加合物以及通过各种DNA修复蛋白修复化疗诱导的DNA损伤的能力有关。在损伤部位加载到DNA上的Rad9-Rad1-HUS1复合物参与检查点激活以及DNA修复。在这里,我们首次探讨了HUS1表达在尿路上皮癌发生中的潜在影响(使用两个人类基底尿路上皮癌细胞系UM-UC-3和HT1197)及其作为预测铂类化疗反应的潜在治疗靶点的作用。通过特异性siRNA实现了对两种细胞系中HUS1表达的特异性抑制,并通过蛋白质免疫印迹进行了验证。为了确定HUS1在细胞增殖调节中的可能重要性,用递增浓度的对照或HUS1 siRNA处理亲本细胞和耐药细胞。在两个人类基底尿路上皮癌细胞系UM-UC-3和HT1197中均观察到HUS1蛋白表达。在顺铂敏感细胞中,敲低HUS1可在顺铂存在的情况下抑制细胞增殖。相反,在耐药细胞中敲低HUS1并未导致对顺铂重新敏感。最后,来自癌症基因组图谱的RNAseq数据提供了证据,表明HUS1表达是UC患者生存不良的一个重要预后因素。总之,HUS1可能在UC中作为一种癌基因起作用,并且可能是细胞对基于顺铂化疗反应的关键决定因素。