Abstract：
Cellular senescence plays a role in several physiological processes including aging, embryonic development, tissue remodeling, and wound healing and is considered one of the main barriers against tumor development. Studies of normal and tumor cells both in culture and in vivo suggest that MYC plays an important role in regulating senescence, thereby contributing to tumor development. We have previously described different common methods to measure senescence in cell cultures and in tissues. Unfortunately, there is no unique marker that unambiguously defines a senescent state, and it is therefore necessary to combine measurements of several different markers in order to assure the correct identification of senescent cells. Here we describe protocols for simultaneous detection of multiple senescence markers in situ, a quantitative fluorogenic method to measure senescence-associated β-galactosidase activity (SA-β-gal), and a new method to detect senescent cells based on the Sudan Black B (SBB) analogue GL13, which is applicable to formalin-fixed paraffin-embedded tissues. The application of these methods in various systems will hopefully shed further light on the role of MYC in regulation of senescence, and how that impacts normal physiological processes as well as diseases and in particular cancer development.
摘要：
细胞衰老在几个生理过程中发挥作用，包括衰老，胚胎发育,组织重塑,和伤口愈合，被认为是对抗肿瘤发展的主要障碍之一。对正常细胞和肿瘤细胞在培养和体内的研究表明，MYC在调节衰老中起着重要作用。从而促进肿瘤的发展。我们先前已经描述了不同的常用方法来测量细胞培养物和组织中的衰老。不幸的是,没有明确定义衰老状态的唯一标记，因此有必要将几种不同标记的测量结果结合起来,以确保衰老细胞的正确鉴定。在这里，我们描述了同时检测多个衰老标记的协议，用于测量衰老相关β-半乳糖苷酶活性（SA-β-gal）的定量荧光方法，以及一种基于苏丹黑B（SBB）类似物GL13的检测衰老细胞的新方法，该方法适用于福尔马林固定的石蜡包埋组织。这些方法在各种系统中的应用有望进一步阐明MYC在衰老调节中的作用。以及这如何影响正常的生理过程以及疾病，特别是癌症的发展。
