Cortisol-producing adrenocortical adenomas (CPAs) are associated with ACTH-independent Cushing’s syndrome and histologically composed of two cellular subtypes: compact (lipid-poor) and clear (lipid-rich) tumor cells. However, the details of hormonal and biological activities of these tumor cells have remained unknown, especially in CPAs. CPAs frequently harbored unique histological features different from those of aldosterone-producing adenomas (APAs) including a senescent phenotype. Therefore, we explored the association between morphological features and the immunoreactivity of steroidogenic enzymes in CPAs with different genotypes and compared them with cellular senescence markers as well as clinicopathological factors of the cases. Hormonal activities (3βHSD, CYP21A, CYP17A1, CYP11B1 and DHEA-ST) and cellular senescence markers (p16, p21 and Ki-67) within different morphological features (clear and compact) were evaluated in 40 CPAs. CPA genotypes (PRKACA, GNAS and CTNNB1) were examined by Sanger sequencing and then compared them with the factors above. p21 immunoreactivity was significantly positively correlated with that of CYP21A (p = 0.0110), CYP17A1 (p = 0.0356) and DHEA-ST (p = 0.0420) but inversely with tumor size (p = 0.0015). CYP21A (p = 0.0016), CYP11B1 (p = 0.0001), CYP17A1 (p < 0.0001) and p16 (p = 0.0137) immunoreactivity were all significantly higher in compact cells than those in clear cells. CYP17A1 (p = 0.0056) and 3βHSD (p = 0.0437) immunoreactivity was significantly higher in PRKACA-mutated than wild type CPAs. p16 immunoreactivity and serum DHEA-S level were both significantly higher in GNAS-mutated than PRKACA-mutated (p = 0.0250) and wild type (p = 0.0180) CPAs. Results of our present study did demonstrate that compact tumor cells were hormonally active and more senescent than clear tumor cells in CPAs. PRKACA- and GNAS-mutated tumor cells were more hormonally active and senescent than those without mutations despite the similar morphological features. We herein proposed a novel histological classification of the tumor cell subtypes based on in situ cortisol excess, genotypes and the status of cell senescence in CPAs.

产生皮质醇的肾上腺皮质腺瘤（CPA）与不依赖ACTH的库欣氏综合征相关，并且在组织学上由两种细胞亚型组成：致密（贫脂）和透明（富脂）肿瘤细胞。但是，这些肿瘤细胞的激素和生物学活性的细节仍然未知，尤其是在CPA中。CPA经常具有与包括衰老表型的醛固酮产生腺瘤（APA）不同的独特组织学特征。因此，我们探讨了不同基因型的CPAs的形态特征与类固醇生成酶的免疫反应性之间的关联，并将其与细胞衰老标记以及病例的临床病理因素进行了比较。激素活性（3βHSD，CYP21A，CYP17A1，CYP11B1和DHEA-ST）和细胞衰老标记（p16，在40个CPA中评估了不同形态特征（清晰和紧凑）中的p21和Ki-67）。CPA基因型（通过Sanger测序检查PRKACA，GNAS和CTNNB1），然后将它们与上述因素进行比较。p21免疫反应性与CYP21A（p = 0.0110），CYP17A1（p = 0.0356）和DHEA-ST（p = 0.0420）呈显着正相关，与肿瘤大小呈负相关（p = 0.0015）。紧密细胞中的CYP21A（p = 0.0016），CYP11B1（p = 0.0001），CYP17A1（p <0.0001）和p16（p = 0.0137）的免疫反应性均显着高于透明细胞。在PRKACA突变后，CYP17A1（p = 0.0056）和3βHSD（p = 0.0437）的免疫反应性显着高于野生型CPA。GNAS突变的p16免疫反应性和血清DHEA-S水平均显着高于PRKACA突变（p = 0.0250）和野生型（p = 0.0180）CPA。我们当前研究的结果确实表明，与CPA中的透明肿瘤细胞相比，致密肿瘤细胞具有荷尔蒙活性，并且衰老程度更高。尽管具有相似的形态特征，但PRKACA和GNAS突变的肿瘤细胞比没有突变的肿瘤细胞具有更强的激素活性和衰老作用。我们在此提出了一种基于原位皮质醇过量，基因型和CPA中细胞衰老状态的肿瘤细胞亚型的新型组织学分类方法。