Kerckaert G A, LeBoeuf R A, Isfort R J
CP & RSD/HSD, The Procter & Gamble Company, Miami Valley Laboratories, Cincinnati, Ohio 45253-8707, USA.
Carcinogenesis. 1996 Sep;17(9):1819-24. doi: 10.1093/carcin/17.9.1819.
We have investigated the effects of modification of the growth media pH on Syrian hamster embryo (SHE) cellular lifespan and transformation. Culturing SHE cells in Dulbecco's modified Eagle's medium (DMEM) at pH 6.7 relative to pH 7.3 results in a 4-fold increase in the lifespan of SHE cells before senescence occurs. This effect was observed both with the wild type SHE cell population and with the morphological transformation sensitive subpopulations derived from the wild type SHE cell mixture (cl9 and cl17). The increase in SHE cell lifespan at pH 6.7 was observed both as an increase in days in culture and an increase in the number of total population doublings before cellular senescence occurred. The tumor promoting agent TPA increased the lifespan of the SHE cell population and one of the MT sensitive subpopulations when these cells were cultured in pH 7.3 medium. Interestingly, treatment of the SHE cell population, and the two MT sensitive SHE cell clones with TPA at culture medium pH of 6.7 reduced the lifespan of each cell type. Finally, culturing SHE cells in medium of pH 6.7 increased the potential of the cells to undergo morphological transformation in response to treatment with carcinogens, demonstrating a potential connection between SHE cell lifespan and carcinogen induced morphological transformation, probably through a mechanism involving a pH induced block in cellular differentiation. We believe this is the first report demonstrating the role of pH in controlling cellular lifespan.
我们研究了生长培养基pH值的改变对叙利亚仓鼠胚胎(SHE)细胞寿命和转化的影响。与pH 7.3相比,在pH 6.7的杜尔贝科改良伊格尔培养基(DMEM)中培养SHE细胞,可使SHE细胞在衰老发生前的寿命增加4倍。在野生型SHE细胞群体以及从野生型SHE细胞混合物中衍生出的对形态转化敏感的亚群(cl9和cl17)中均观察到了这种效应。在pH 6.7时SHE细胞寿命的增加表现为培养天数的增加以及细胞衰老发生前总群体倍增数的增加。当这些细胞在pH 7.3的培养基中培养时,肿瘤促进剂TPA增加了SHE细胞群体以及其中一个对甲基胆蒽(MT)敏感的亚群的寿命。有趣的是,在培养基pH为6.7时用TPA处理SHE细胞群体以及两个对MT敏感的SHE细胞克隆,会缩短每种细胞类型的寿命。最后,在pH 6.7的培养基中培养SHE细胞增加了细胞对致癌物处理发生形态转化的潜力,这表明SHE细胞寿命与致癌物诱导的形态转化之间可能存在联系,可能是通过一种涉及pH诱导细胞分化阻滞的机制。我们认为这是第一份证明pH在控制细胞寿命中作用的报告。