New biomarkers probing depth of cell senescence assessed by laser scanning cytometry.
New biomarkers probing depth of cell senescence assessed by laser scanning cytometry.
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DOI：
10.1002/cyto.a.20983
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发表时间：
2010-11
期刊：
影响因子：
3.7
通讯作者：
Darzynkiewicz, Zbigniew
中科院分区：
文献类型：
--
作者：
Zhao, Hong;Halicka, H. Dorota;Traganos, Frank;Jorgensen, Ellen;Darzynkiewicz, Zbigniew
关键词：
激光扫描细胞仪（LSC）的成像分析能力已被用于评估被认为是典型的衰老表型的形态学特征。衰老细胞的特征性“变平”表现为dna相关荧光[4,6-二氨基-2-苯基吲哚（DAPI）]染色密度（最大像素强度）下降，同时细胞核大小（面积）增加。最大像元/核面积比值的下降是比最大像元或核面积变化更敏感的衰老标志物。在衰老细胞的培养中，LSC记录的生长平台期饱和细胞密度显著降低，这也可以作为一个额外的标记。细胞周期蛋白依赖性激酶抑制剂p21WAF1、p27KIP1和γ - h2ax的诱导和DNA损伤反应ATM标记的激活平行于DNA/DAPI最大像素和核面积。这些生物标志物指数通过报告它们作为各自对照的一部分来定量表达。研究不同浓度米托蒽醌（Mxt）和曲古抑素A在不同时间对A549和WI-38细胞的作用，评估细胞衰老的程度（深度）。还评估了2-脱氧-d-葡萄糖（降低代谢细胞活性的物质）对Mxt诱导的衰老深度的影响。还研究了细胞合成RNA（含5-乙基尿苷）导致生长失衡与细胞衰老之间的关系。数据表明，LSC对细胞属性的形态计量学分析提供了一种有吸引力的工具来检测细胞衰老并测量其程度，特别是在评估增强或减弱这一过程的因素的影响方面。这种方法很重要，因为有证据表明，细胞衰老不仅是机体衰老的基础生物学过程，而且还阻碍诱导多能干细胞的形成，为肿瘤转化提供屏障，是实体瘤治疗期间诱导生殖细胞死亡的主要机制。©2010国际细胞术进步学会。
The imaging analytical capabilities of laser scanning cytometer (LSC) have been used to assess morphological features considered to be typical of the senescent phenotype. The characteristic “flattening” of senescent cells was reflected by the decline in the density of staining (intensity of maximal pixel) of DNA-associated fluorescence [4,6-diamidino-2-phenylindole (DAPI)] paralleled by an increase in nuclear size (area). The decrease in ratio of maximal pixel to nuclear area was even more sensitive senescence biomarker than the change in maximal pixel or nuclear area, each alone. The saturation cell density at plateau phase of growth recorded by LSC was found to be dramatically decreased in cultures of senescent cells, thereby also serving as an additional marker. The induction of cyclin dependent kinase inhibitors p21WAF1 and p27KIP1 and γH2AX and activation of ATM markers of DNA damage response were measured in parallel with DNA/DAPI maximal pixel and nuclear area. These biomarker indices were expressed in quantitative terms by reporting them as a fraction of the respective controls. The effect of treatment of A549 and WI-38 cells with different concentrations of mitoxantrone (Mxt) and trichostatin A for various time periods was studied to assess the degree (depth) of cell senescence. Also assessed was the effect of 2-deoxy-d-glucose, the agent attenuating metabolic cell activity, on the depth of senescence induced by Mxt. A relationship between the ability of cells to synthesize RNA (incorporate 5-ethynyluridine) that leads to growth imbalance and induction of cell senescence was also studied. The data show that morphometric analysis of cellular attributes by LSC offers an attractive tool to detect cell senescence and measure its degree particularly in assessing effects of the factors that enhance or attenuate this process. This methodology is of importance in light of the evidence that cellular senescence is not only a biological process that is fundamental for organismal aging but also impedes formation of induced-pluripotent stem cells providing the barrier for neoplastic transformation and is the major mechanism of induction of reproductive cell death during treatment of solid tumors. © 2010 International Society for Advancement of Cytometry.
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DOI：
10.1073/pnas.92.20.9363
发表时间：
1995-09-26
影响因子：
11.1
作者：
DIMRI, GP;LEE, XH;CAMPISI, J
通讯作者：
CAMPISI, J
影响因子：
4.3
作者：
Korotchkina, Lioubov G.;Demidenko, Zoya N.;Blagosklonny, Mikhail V.
通讯作者：
Blagosklonny, Mikhail V.
影响因子：
4.3
作者：
Blagosklonny, Mikhail V.
通讯作者：
Blagosklonny, Mikhail V.
影响因子：
4.3
作者：
Blagosklonny, Mikhail V.
通讯作者：
Blagosklonny, Mikhail V.
DOI：
10.1186/bcr2583
发表时间：
2010
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Karimi-Busheri F;Rasouli-Nia A;Mackey JR;Weinfeld M
通讯作者：
Weinfeld M