Screening of senescence-associated genes with specific DNA array reveals the role of IGFBP-3 in premature senescence of human diploid fibroblasts
Florence Debacq‐ChainiauxThierry PascalEmmanuelle BoilanCoralie BastinÉmilie BauwensOlivier Toussaint
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Senescence
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Abstract. The triploid V79 cells are stable under usual culture conditions, and do not revert to being diploid. Here, the triploid–diploid transition of triploid V79 cells has been successfully induced in suspension culture in culture dishes with untreated surfaces. The diploid cells began to appear in a population of triploid V79 cells cultured under these conditions for 4 weeks. All of the triploid cells were transformed to diploid through subsequent monolayer culture for 5 weeks. It was confirmed that the revertant diploid cells had the same characteristics as original diploid V79 cells, with respect to DNA histograms, cell volume and chromosome number. Thus, it seems that suspension culturing is an important factor that induces the triploid–diploid transition.
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Senescence
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Senescence
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Two strains of diploid fibroblasts were derived from green monkey fetal lungs and their main properties were studied. These strains were shown not to differ principally from the well studied human diploid fibroblasts. The derived cultures are free from contaminating viruses. The titers of attenuated poliomyelitis virus strains achieved in green monkey diploid cells are comparable to those in primary cultures of green monkey kidneys. The possiblity of using green monkey diploid fibroblasts in virological practice is discussed.
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Saccharomyces cerevisiae was grown in a rich medium under the conditions of "quasi-continuous" cultivation and, after 200-300 generations, its diploid cells almost completely displaced haploid cells from the original mixed "haploid-diploid" population where the ratio between diploid and haploid strains was either 1:1 or 1:100. The cultivation at 40 degrees C did not change the relative competitive ability of haploids and diploids. When cells were cultivated in a rich medium at 6 degrees C or in a minimal medium at 30 degrees C, none of the strains showed an advantage over others for about 200 generations. Haploid cells had an advantage over diploid cells during "quasi-continuous" growth in the minimal medium at 30 degrees C. When the temperature was elevated to 40 degrees C, diploid cells displaced haploid cells from the mixed population. No advantage was found for diploid or haploid cells grown in a medium with an elevated KCl content (1.5 M). Haploid cells had an advantage over diploid cells when Pichia pinus was cultivated in a minimal medium. The results are discussed using the hypothesis about the diploid phase being fixed in the course of biological evolution.
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DNA damage is the major cause of senescence and apoptosis; however, the manner by which DNA-damaged cells become senescent remains unclear. We demonstrate that DNA damage leads to a greater level of senescence rather than apoptosis in DBC1-deficient cells. In addition, we show that BLM becomes degraded during DNA damage, which induces p21 expression and senescence. DBC1 binds to and shields BLM from degradation, thus suppressing senescence. ML216 promotes DBC1–BLM interaction, which aids in the preservation of BLM following DNA damage and suppresses senescence. ML216 enhances pulmonary function by lowering the levels of senescence and fibrosis in both aged mice and a mouse model of bleomycin-induced idiopathic pulmonary fibrosis. Our data reveal a unique mechanism preventing DNA-damaged cells from becoming senescent, which may be regulated by the use of ML216 as a potential treatment for senescence-related diseases.
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A new human diploid fibroblast-like cell strain has been established from dermal tissue of a 2 months old female embryo. This cell strain showed diploid karyo type and has a finite life span of 95－100 generations. No spontaneous transformation was observed during the whole life span in vitro. This cell strain is susceptible to SV40 infection and can be transformed easily by it.
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To examine cellular changes after diploid-tetraploid transformation in mammalian cells, tetraploid Meth-A cells, which were established from diploid cell line, were compared to the parent diploid cells for serum dependence of growth. Tetraploid Meth-A cells showed a high tolerance for low serum concentrations and could proliferate in a serum-free medium, differing from the parent diploid cells. It was suggested that diploid-tetraploid transition of Meth-A cells resulted in induction of a new cellular function, being that hyperploid cells can proliferate in sever circumstance for nutriments.
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Abstract Replicative senescence is a process in which normal proliferative cells lose replicative potential and acquire biomarkers of cellular senescence as a result of serial passages in culture instead of chronological time. Oxidative stress‐induced premature senescence is the long‐term appearance of the biomarkers of replicative senescence after exposure of human diploid cells to sublethal stress. Cytokine‐induced premature senescence is the long‐term appearance of biomarkers of replicative senescence after repeated exposure of human diploid fibroblasts to proinflammatory cytokines.
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