Gadepalli Venkat S, Vaughan Catherine, Rao Raj R
Integrative Life Sciences, Department of Life Sciences, Virginia Commonwealth University, Richmond, VA, USA.
Methods Mol Biol. 2013;962:183-91. doi: 10.1007/978-1-62703-236-0_15.
Cancer stem cells possess the ability to self-renew and differentiate into specific cells found in tumor types, a characteristic feature of normal multipotent stem cells. These cells harbor within the bulk of tumors and if the tumor suppressor p53 is mutated in these cells, can be more likely to cause relapse and metastasis by giving rise to new tumors. This new paradigm of oncogenesis has been observed in various cancers, including lung cancer. Determining the interaction of critical cellular pathways in the ontogeny of lung tumors is expected to lead to identification of molecular targets for effective therapeutic strategies. To achieve this, it is important to characterize and dissect the differences between the cancer cells with aberrant stem cell like properties and normal multipotent stem cells that contribute to regeneration. This could be accomplished by using cell surface markers unique for certain cell types by employing techniques such as flow cytometry and magnetic bead isolation. This chapter summarizes the isolation process of the resident stem cell Sca1 (+ve), CD-45 (-ve), and CD-31 (-ve) populations for its potential use in assessing correlations between specific p53 gain of function phenotypes in different murine lung cancer models.
癌症干细胞具有自我更新的能力,并能分化为肿瘤类型中发现的特定细胞,这是正常多能干细胞的一个特征。这些细胞存在于肿瘤主体中,如果肿瘤抑制因子p53在这些细胞中发生突变,就更有可能通过产生新的肿瘤而导致复发和转移。这种肿瘤发生的新范式已在包括肺癌在内的各种癌症中被观察到。确定肺肿瘤发生过程中关键细胞通路的相互作用,有望找到有效治疗策略的分子靶点。为了实现这一点,重要的是要表征和剖析具有异常干细胞样特性的癌细胞与有助于再生的正常多能干细胞之间的差异。这可以通过使用特定细胞类型特有的细胞表面标志物,采用流式细胞术和磁珠分离等技术来完成。本章总结了驻留干细胞Sca1(阳性)、CD - 45(阴性)和CD - 31(阴性)群体的分离过程,其潜在用途是评估不同小鼠肺癌模型中特定p53功能获得表型之间的相关性。