背景:ZEB1是一种参与上皮-间质转化(EMT)的核心转录因子,与侵袭性癌细胞行为有关,治疗抗性,以及各种肿瘤类型的不良预后。同样,CD73的表达和活性,CD73是一种与腺苷生成有关的外核苷酸酶,是肿瘤恶性程度的重要标志。越来越多的证据表明,EMT和腺苷途径是错综复杂的联系,并在癌症发展中起关键作用。因此,考虑CD73和ZEB1对肿瘤进展的影响,本研究重点探讨CD73和ZEB1之间的相关性.
方法:我们采用CRISPR/Cas9技术沉默甲状腺乳头状癌细胞系中CD73的表达。这些相同的细胞经历ZEB1非编码RNA调控的报道分子的慢病毒转导。我们使用划痕测定和细胞速度和极性分析对细胞迁移进行了研究。此外,我们通过流式细胞术和免疫细胞化学检查了ZEB1报告表达,通过蛋白质印迹分析补充蛋白质定量。对于进一步的见解,我们将代表不同EMT状态的基因签名应用于来自癌症基因组图谱的甲状腺乳头状癌样本的RNA-seq表达分析中.
结果:沉默CD73表达导致甲状腺乳头状癌衍生细胞系中ZEB1非编码RNA调控报告表达减少。此外,它还减轻ZEB1蛋白表达。此外,CD73和ZEB1的表达与对细胞迁移至关重要的细胞形态特征的改变相关,促进细胞极性指数和细胞迁移速度的增加。RNA-seq分析显示NT5E(CD73)在有BRAF突变的样品中表达较高,伴随着部分EMT/杂合状态签名表达的流行。
结论:总的来说,我们的发现表明CD73表达和/或活性与非编码RNA对ZEB1的转录后调控之间存在关联,表明其缺席的减少。需要进一步的研究来阐明CD73和ZEB1之间的关系,并有可能在不久的将来将其作为癌症治疗的治疗替代方案。
ZEB1, a core transcription factor involved in epithelial-mesenchymal transition (EMT), is associated with aggressive cancer cell behavior, treatment resistance, and poor prognosis across various tumor types. Similarly, the expression and activity of CD73, an ectonucleotidase implicated in adenosine generation, is an important marker of tumor malignancy. Growing evidence suggests that EMT and the adenosinergic pathway are intricately linked and play a pivotal role in cancer development. Therefore, this study focuses on exploring the correlations between CD73 and ZEB1, considering their impact on tumor progression.
We employed CRISPR/Cas9 technology to silence CD73 expression in cell lines derived from papillary thyroid carcinoma. These same cells underwent lentiviral transduction of a reporter of ZEB1 non-coding RNA regulation. We conducted studies on cell migration using scratch assays and analyses of cellular speed and polarity. Additionally, we examined ZEB1 reporter expression through flow cytometry and immunocytochemistry, complemented by Western blot analysis for protein quantification. For further insights, we applied gene signatures representing different EMT states in an RNA-seq expression analysis of papillary thyroid carcinoma samples from The Cancer Genome Atlas.
Silencing CD73 expression led to a reduction in ZEB1 non-coding RNA regulation reporter expression in a papillary thyroid carcinoma-derived cell line. Additionally, it also mitigated ZEB1 protein expression. Moreover, the expression of CD73 and ZEB1 was correlated with alterations in cell morphology characteristics crucial for cell migration, promoting an increase in cell polarity index and cell migration speed. RNA-seq analysis revealed higher expression of NT5E (CD73) in samples with BRAF mutations, accompanied by a prevalence of partial-EMT/hybrid state signature expression.
Collectively, our findings suggest an association between CD73 expression and/or activity and the post-transcriptional regulation of ZEB1 by non-coding RNA, indicating a reduction in its absence. Further investigations are warranted to elucidate the relationship between CD73 and ZEB1, with the potential for targeting them as therapeutic alternatives for cancer treatment in the near future.