PO.MCB01.01 · 分子与细胞生物学
鉴定氯倍他索诱导UMSCV-4外阴癌细胞进入静息期过程中细胞周期调控蛋白的变化
Identifying changes in cell cycle regulatory proteins during clobetasol-induced quiescence in UMSCV-4 vulvar cancer cells
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
静息是一种可逆状态,细胞在此状态下响应应激而暂时退出细胞周期,并具有在给定应激解除后重新进入周期的能力。越来越多的证据表明,静息的癌细胞可能是长期肿瘤休眠和晚期复发的基础。我们发现外阴鳞状细胞癌细胞系UMSCV-4中的一个亚群在暴露于皮质类固醇氯倍他索(clob)后进入静息期。去除clob后,这些细胞重新进入细胞周期。为表征这一过程,我们使用BrdU掺入定量了增殖的变化,并评估了clob撤除后细胞周期重新进入的时间。我们假设clob处理会下调关键的促细胞周期蛋白,同时上调细胞周期抑制因子,且这些变化会在去除clob后三天内逆转,与DNA合成增加和有丝分裂象重新出现相一致。使用Western blot分析,我们检测了细胞周期促进因子(cyclin D1、cyclin D3、CDK2、CDK4、CDK6)和抑制因子(p21Waf1/Cip1和p18INK4C)的表达。RNA seq分析正被用于评估相应的mRNA丰度变化,并鉴定与clob诱导的阻滞相关的其他调控因子。总之,这些研究旨在阐明氯倍他索促进外阴癌细胞进入静息期的分子机制。
查看英文原文 English abstract
Quiescence is a reversible state in which cells temporarily exit the cell cycle in response to stress, with the capacity to re-enter the cycle once the given stress is relieved. Growing evidence suggests that quiescent cancer cells may underlie long-term tumor dormancy and late relapse. We found that a subpopulation of the vulvar squamous cell carcinoma line UMSCV-4 enters quiescence following exposure to the corticosteroid clobetasol (clob). Upon removal of clob, these cells re-enter the cell cycle. To characterize this process, we quantified changes in proliferation using BrdU incorporation and assessed the timing of cell cycle re-entry after clob withdrawal. We hypothesized that clob treatment would downregulate key cell-cycle-promoting proteins while upregulating cell-cycle inhibitors, and that these changes would reverse within three days of clob removal, coinciding with increased DNA synthesis and the reappearance of mitotic figures. Using Western blot analysis, we examined the expression of cell-cycle promoters (cyclin D1, cyclin D3, CDK2, CDK4, CDK6) and inhibitors (p21Waf1/Cip1 and p18INK4C). RNA seq analysis is being used to evaluate corresponding changes in mRNA abundance and to identify additional regulators associated with clob-induced arrest. Together, these studies aim to define the molecular mechanisms by which clobetasol promotes quiescence in vulvar cancer cells.
利益披露 Disclosure
C. M. Marsello, None..
T. A. Trojanowski, None..
J. E. Lewis, None.