Background:Metastatic prostate cancer (mPCa) is marked by heterogeneity and therapy resistance, which arise from prolonged therapy regimens. This heterogeneity is reflected in various morphologic and genetic characteristics, biomarker expression, and other molecular mechanisms, thereby contributing to the complexity of the disease.Methods:To investigate tumor heterogeneity, the effects of androgen targeting therapy (ADT) on single-cell PSA secretion was assessed by analyzing the prostate cancer cell lines using a modified ELISpot platform. The FACS and cytospin techniques were employed to understand the influence of the cell cycle on PSA secretion patterns. Additionally, a proteome array was used to identify potential biomarkers from different PCa cell lines with varying metastatic potential.Results:Among the various PCa cell lines examined, PSA expression and secretion could be visualized only from the LNCaPs. PSA secretion from circulating tumor cells (CTCs) further confirmed the validity of this assay. These LNCaPs exhibited heterogeneity in single-cell intracellular and extracellular PSA expression and in their ADT responses. LNCaPs in the G1 phase showed higher PSA secretion than in the S or G2/M phase. Apart from PSA, Cathepsin D, Progranulin, IL-8, Serpin E1, and Enolase 2 were identified as secretome markers from the metastatic PCa cell lines.Conclusions:We observed variability in PSA secretion in LNCaP in response to anti-androgen treatment and a cell cycle-dependent secretion pattern. The notable presence of Progranulin and Cathepsin D in metastatic cell lines makes them promising candidates for use in multiplexing and single-cell platforms, potentially advancing our understanding and treatment of this disease.
背景：转移性前列腺癌（mPCa）具有异质性和治疗抵抗性，这些特性源于长期的治疗方案。这种异质性体现在多种形态和遗传特征、生物标志物表达以及其他分子机制上，从而增加了疾病的复杂性。 方法：为研究肿瘤异质性，我们通过改良的ELISpot平台分析前列腺癌细胞系，评估了雄激素靶向治疗（ADT）对单细胞前列腺特异性抗原（PSA）分泌的影响。采用流式细胞术和细胞离心涂片技术，探究细胞周期对PSA分泌模式的影响。此外，利用蛋白质组芯片技术，从具有不同转移潜能的前列腺癌细胞系中鉴定潜在的生物标志物。 结果：在所检测的各种前列腺癌细胞系中，仅LNCaP细胞系显示出PSA的表达和分泌。循环肿瘤细胞（CTCs）的PSA分泌进一步验证了该检测方法的有效性。这些LNCaP细胞在单细胞内外PSA表达及对ADT的反应上表现出异质性。处于G1期的LNCaP细胞比处于S期或G2/M期的细胞分泌更多的PSA。除PSA外，组织蛋白酶D、颗粒蛋白前体、IL-8、丝氨酸蛋白酶抑制剂E1和烯醇化酶2被鉴定为转移性前列腺癌细胞系的分泌组标志物。 结论：我们观察到LNCaP细胞在抗雄激素治疗下PSA分泌存在变异性，并呈现出细胞周期依赖性的分泌模式。转移性细胞系中颗粒蛋白前体和组织蛋白酶D的显著存在，使其成为用于多重检测和单细胞平台的有前景的候选标志物，有望推动对该疾病的理解和治疗进展。
肿瘤(癌症)患者之家