Turai Péter István, Nyirő Gábor, Borka Katalin, Micsik Tamás, Likó István, Patócs Attila, Igaz Peter
Department of Endocrinology, ENS@T Research Center of Excellence, Faculty of Medicine, Semmelweis University, H-1083 Budapest, Hungary.
Department of Internal Medicine and Oncology, Faculty of Medicine, Semmelweis University, H-1083 Budapest, Hungary.
Cancers (Basel). 2022 Sep 2;14(17):4313. doi: 10.3390/cancers14174313.
Differentiation of adrenocortical adenoma (ACA) and carcinoma (ACC) is often challenging even in the histological analysis. Circular RNAs (circRNAs) belonging to the group of non-coding RNAs have been implicated as relevant factors in tumorigenesis. Our aim was to explore circRNA expression profiles in adrenocortical tumors by next-generation sequencing followed by RT-qPCR validation. Archived FFPE (formalin-fixed, paraffin embedded) including 8 ACC, 8 ACA and 8 normal adrenal cortices (NAC) were used in the discovery cohort. For de novo and known circRNA expression profiling, a next-generation sequencing platform was used. CIRI2, CircExplorer2, AutoCirc bioinformatics tools were used for the discovery of circRNAs. The top five most differentially circRNAs were measured by RT-qPCR in an independent validation cohort (10 ACC, 8 ACA, 8 NAC). In silico predicted, interacting microRNAs potentially sponged by differentially expressed circRNAs were studied by individual RT-qPCR assays. We focused on overexpressed circRNAs here. Significantly differentially expressed circRNAs have been revealed between the cohorts by NGS. Only could be confirmed to be significantly overexpressed in ACC, ACA vs. NAC samples by RT-qPCR. We could not observe microRNA expression changes fully corresponding to our sponging hypothesis. To the best of our knowledge, our study is the first to investigate circRNAs in adrenocortical tumors. Further studies are warranted to explore their biological and diagnostic relevance.
即使在组织学分析中，肾上腺皮质腺瘤（ACA）和癌（ACC）的鉴别也常常具有挑战性。属于非编码RNA组的环状RNA（circRNA）已被认为是肿瘤发生中的相关因素。我们的目的是通过下一代测序，随后进行RT-qPCR验证，来探索肾上腺皮质肿瘤中的circRNA表达谱。发现队列中使用了存档的FFPE（福尔马林固定、石蜡包埋）样本，包括8例ACC、8例ACA和8例正常肾上腺皮质（NAC）。对于从头和已知的circRNA表达谱分析，使用了下一代测序平台。使用CIRI2、CircExplorer2、AutoCirc生物信息学工具来发现circRNA。在一个独立的验证队列（10例ACC、8例ACA、8例NAC）中，通过RT-qPCR测量了差异最大的前五种circRNA。通过单独的RT-qPCR分析研究了在计算机上预测的、可能被差异表达的circRNA海绵化的相互作用微小RNA。我们在此关注过表达的circRNA。通过NGS在队列之间发现了显著差异表达的circRNA。通过RT-qPCR仅能确认在ACC、ACA与NAC样本中显著过表达。我们没有观察到微小RNA表达变化完全符合我们的海绵化假说。据我们所知，我们的研究是首次调查肾上腺皮质肿瘤中的circRNA。有必要进行进一步研究以探索它们的生物学和诊断相关性。
