Laboratory of Human Carcinogenesis, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):2195-200. doi: 10.1073/pnas.0909390107. Epub 2010 Jan 13.
Normal tissue homeostasis is maintained through asymmetric cell divisions that produce daughter cells with differing self-renewal and differentiation potentials. Certain tumor cell subfractions can self-renew and repopulate the heterogeneous tumor bulk, suggestive of asymmetric cell division, but an equally plausible explanation is that daughter cells of a symmetric division subsequently adopt differing cell fates. Cosegregation of template DNA during mitosis is one mechanism by which cellular components are segregated asymmetrically during cell division in fibroblast, muscle, mammary, intestinal, and neural cells. Asymmetric cell division of template DNA in tumor cells has remained elusive, however. Through pulse-chase experiments with halogenated thymidine analogs, we determined that a small population of cells within human lung cancer cell lines and primary tumor cell cultures asymmetrically divided their template DNA, which could be visualized in single cells and in real time. Template DNA cosegregation was enhanced by cell-cell contact. Its frequency was density-dependent and modulated by environmental changes, including serum deprivation and hypoxia. In addition, we found that isolated CD133(+) lung cancer cells were capable of tumor cell repopulation. Strikingly, during cell division, CD133 cosegregated with the template DNA, whereas the differentiation markers prosurfactant protein-C and pan-cytokeratins were passed to the opposing daughter cell, demonstrating that segregation of template DNA correlates with lung cancer cell fate. Our results demonstrate that human lung tumor cell fate decisions may be regulated during the cell division process. The characterization and modulation of asymmetric cell division in lung cancer can provide insight into tumor initiation, growth, and maintenance.
正常组织稳态通过不对称细胞分裂来维持,这种分裂产生具有不同自我更新和分化潜力的子细胞。某些肿瘤细胞亚群可以自我更新并重新填充异质性肿瘤块,这表明存在不对称细胞分裂,但同样合理的解释是,对称分裂的子细胞随后会采用不同的细胞命运。有丝分裂过程中模板 DNA 的共分离是一种机制,通过该机制,在成纤维细胞、肌肉、乳腺、肠道和神经细胞中,细胞成分在细胞分裂过程中被不对称地分离。然而,肿瘤细胞中模板 DNA 的不对称分裂仍然难以捉摸。通过氚化胸腺嘧啶类似物的脉冲追踪实验,我们确定人肺癌细胞系和原代肿瘤细胞培养物中的一小部分细胞不对称地分裂了它们的模板 DNA,可以在单细胞和实时中观察到。模板 DNA 的共分离受细胞间接触的增强。其频率与环境变化(包括血清剥夺和缺氧)有关,并受其调节。此外,我们发现分离的 CD133(+)肺癌细胞能够重新填充肿瘤细胞。引人注目的是,在细胞分裂过程中,CD133 与模板 DNA 共分离,而前表面活性剂蛋白-C 和泛细胞角蛋白等分化标记物则传递到对面的子细胞,这表明模板 DNA 的分离与肺癌细胞命运相关。我们的研究结果表明,人类肺癌细胞命运决定可能在细胞分裂过程中受到调节。肺癌中不对称细胞分裂的特征和调节可以深入了解肿瘤的起始、生长和维持。