Table_2_Epidermal Growth Factor Is Essential for the Maintenance of Novel Prostate Epithelial Cells Isolated From Patient-Derived Organoids.DOCX

Prostate cancer (PCa) is the second leading cause of cancer-related mortality and morbidity among males worldwide. Deciphering the biological mechanisms and molecular pathways involved in PCa pathogenesis and progression has been hindered by numerous technical limitations mainly attributed to the limited number of cell lines available, which do not recapitulate the diverse phenotypes of clinical disease. Indeed, PCa has proven problematic to establish as cell lines in culture due to its heterogeneity which remains a challenge, despite the various in vitro and in vivo model systems available. Growth factors have been shown to play a central role in the complex regulation of cell proliferation among hormone sensitive tumors, such as PCa. Here, we report the isolation and characterization of novel patient-derived prostate epithelial (which we named as AUB-PrC) cells from organoids culture system. We also assessed the role of epidermal growth factor (EGF) in culturing those cells. We profiled the AUB-PrC cells isolated from unaffected and tumor patient samples via depicting their molecular and epithelial lineage features through immunofluorescence staining and quantitative real-time PCR (qRT-PCR), as well as through functional assays and transcriptomic profiling through RNA sequencing. In addition, by optimizing a previously established prostate organoids culture system, we were able to grow human prostate epithelial cells using growth medium and EGF only. With these data collected, we were able to gain insight at the molecular architecture of novel human AUB-PrC cells, which might pave the way for deciphering the mechanisms that lead to PCa development and progression, and ultimately improving prognostic abilities and treatments.

前列腺癌（PCa）是全球范围内男性癌症相关死亡和发病率第二高的重要原因。解析前列腺癌发病机制和进展过程中涉及的生物学机制与分子通路，受到了诸多技术限制的阻碍，这些限制主要归因于可用的细胞系数量有限，这些细胞系无法重现临床疾病的多样性表型。实际上，由于前列腺癌的异质性，尽管存在各种体外和体内模型系统，其作为细胞系在培养中的建立一直是一个难题。生长因子已被证明在激素敏感型肿瘤（如前列腺癌）中细胞增殖的复杂调节中起着核心作用。在本研究中，我们报告了从器官oids培养系统中分离和鉴定出新型患者来源的前列腺上皮细胞（我们将其命名为AUB-PrC）。我们还评估了表皮生长因子（EGF）在培养这些细胞中的作用。通过免疫荧光染色和定量实时PCR（qRT-PCR），以及通过功能实验和RNA测序进行转录组分析，我们描绘了从未受影响和肿瘤患者样本中分离的AUB-PrC细胞的分子和上皮谱系特征。此外，通过优化先前建立的前列腺器官oids培养系统，我们仅使用生长培养基和EGF就能培养出人前列腺上皮细胞。收集这些数据后，我们得以洞察新型人类AUB-PrC细胞的分子结构，这可能会为解开导致前列腺癌发展和进展的机制提供途径，并最终提高预后能力和治疗方案。