背景:肾上腺皮质癌(ACC)是一种极其罕见的,侵袭性肿瘤,几乎没有有效的治疗选择或药物。Mitotane(Mtn),这是唯一授权的治疗药物,1970年问世,尽管有严重的不良反应和高复发率,但仍然是ACC的唯一一线治疗方法。
方法:通过对癌症基因组图谱(TCGA)中ACC数据集的计算机模拟分析,我们确定细胞周期蛋白依赖性激酶-1(CDK1)的高表达水平与ACC的不良临床结局显著相关.进行体外和体内实验以通过ACC细胞中功能的获得和丧失测定来评估CDK1在ACC进展中的作用。筛选CDK1抑制剂以鉴定用于治疗ACC的潜在候选物。RNA测序,免疫共沉淀,和免疫荧光分析用于阐明机制。
结果:在ACC细胞系中过表达CDK1促进增殖并诱导上皮-间质转化(EMT),而CDK1表达的敲除抑制ACC细胞系的生长。CDK1抑制剂,葫芦素E(CurE),在体内和体外均具有最佳的抑制作用,具有良好的时间和剂量依赖性活性。CurE对裸鼠ACC异种移植物的抑制作用大于米托坦,无明显不良反应。最重要的是,CurE和米托坦联合治疗几乎完全消除了ACC肿瘤。关于机制,CDK1通过Slug和Twist促进ACC细胞的EMT,并通过与UBE2C和AURKA/B的相互作用将ACC细胞锁定到G2/M检查点。CDK1还调节焦亡,凋亡,和ACC细胞的坏死(PANoptosis)通过与PANopsome以ZBP1依赖性方式结合。
结论:CDK1可以通过调节EMT作为ACC的重要治疗靶点,G2/M检查点,和全景。因此,CurE可能是ACC治疗的潜在候选药物,具有良好的安全性和有效性。这将满足ACC患者的巨大需求。
BACKGROUND: Adrenocortical carcinoma (ACC) is an extremely rare, aggressive tumor with few effective therapeutic options or drugs.
Mitotane (Mtn), which is the only authorized therapeutic drug, came out in 1970 and is still the only first-line treatment for ACC in spite of serious adverse reaction and a high recurrence rate.
METHODS: By in silico analysis of the ACC dataset in the cancer genome atlas (TCGA), we determined that high expression levels of cyclin-dependent kinase-1 (CDK1) were significantly related to the adverse clinical outcomes of ACC. In vitro and in vivo experiments were performed to evaluate the role of CDK1 in ACC progression through gain and loss of function assays in ACC cells. CDK1 inhibitors were screened to identify potential candidates for the treatment of ACC. RNA sequencing, co-immunoprecipitation, and immunofluorescence assays were used to elucidate the mechanism.
RESULTS: Overexpression of CDK1 in ACC cell lines promoted proliferation and induced the epithelial-to-mesenchymal transition (EMT), whereas knockdown of CDK1 expression inhibited growth of ACC cell lines. The CDK1 inhibitor, cucurbitacin E (CurE), had the best inhibitory effect with good time-and dose-dependent activity both in vitro and in vivo. CurE had a greater inhibitory effect on ACC xenografts in nude mice than
mitotane, without obvious adverse effects. Most importantly, combined treatment with CurE and
mitotane almost totally eliminated ACC tumors. With respect to mechanism, CDK1 facilitated the EMT of ACC cells via Slug and Twist and locked ACC cells into the G2/M checkpoint through interaction with UBE2C and AURKA/B. CDK1 also regulated pyroptosis, apoptosis, and necroptosis (PANoptosis) of ACC cells through binding with the PANoptosome in a ZBP1-dependent way.
CONCLUSIONS: CDK1 could be exploited as an essential therapeutic target of ACC via regulating the EMT, the G2/M checkpoint, and PANoptosis. Thus, CurE may be a potential candidate drug for ACC therapy with good safety and efficacy, which will meet the great need of patients with ACC.