Early loss of proliferative potential of human peritoneal mesothelial cells in culture:: the role of p16INK4a-mediated premature senescence
Early loss of proliferative potential of human peritoneal mesothelial cells in culture:: the role of p16INK4a-mediated premature senescence
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DOI：
10.1152/japplphysiol.01086.2005
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发表时间：
2006-03-01
影响因子：
3.3
通讯作者：
Witowski, J
中科院分区：
文献类型：
--
作者：
Ksiazek, K;Piwocka, K;Witowski, J
关于细胞衰老的机制，我们已经了解了很多。导致衰老的途径似乎有所不同，这取决于细胞类型和细胞培养条件。在这方面，人们对人腹膜间皮细胞（HPMC）的衰老知之甚少。先前的研究在报道的HPMC增殖寿命上有显著差异。因此，在本研究中，我们研究了HPMC如何在通常用于HPMC培养的条件下进入衰老状态。从大网膜中分离出HPMC，并逐渐衰老。对培养物的生长速度、衰老标志物的存在、细胞周期抑制剂的激活和氧化应激进行了评估。发现HPMC在衰老前平均达到6倍的种群数量。终末生长停滞与Ki67抗原表达降低、G1期细胞百分比增加、早期群体双倍水平cDNA-1 mRNA表达减少以及衰老相关β -半乳糖苷酶的存在有关。与早期传代细胞相比，晚期传代细胞p16（INK4a）的表达增加，而p21（Cip1）的表达没有增加。此外，这些细胞产生更多的活性氧，并显示出氧化修饰DNA（8-羟基-2'-脱氧鸟苷）的增加。这些结果表明，网膜源性HPMC的早发性衰老可能与氧化应激诱导的p16INK4a上调有关。
Much has been learned about the mechanisms underlying cellular senescence. The pathways leading to senescence appear to vary, depending on the cell type and cell culture conditions. In this respect, little is known about senescence of human peritoneal mesothelial cells (HPMC). Previous studies have significantly differed in the reported proliferative lifespan of HPMC. Therefore, in the present study, we have examined how HPMC enter state of senescence under conditions typically used for HPMC culture. HPMC were isolated from omentum and grown into senescence. The cultures were assessed for the growth rate, the presence of senescence markers, activation of cell- cycle inhibitors, and the oxidative stress. HPMC were found to reach, on average, six population doublings before senescence. The terminal growth arrest was associated with decreased expression of Ki67 antigen, increased percentage of cells in the G1 phase, reduced early population doubling level cDNA-1 mRNA expression, and the presence of senescence-associated beta-galactosidase. Compared with early-passage cells, the late-passage HPMC exhibited increased expression of p16(INK4a) but not of p21(Cip1). In addition, these cells generated more reactive oxygen species and displayed increased presence of oxidatively modified DNA (8-hydroxy-2'-deoxyguanosine). These results demonstrate that early onset of senescence in omentum-derived HPMC may be associated with oxidative stress-induced upregulation of p16INK4a.