Cloning and Identification of Genes That Associate with Mammalian Replicative Senescence

doi:10.1006/excr.1998.3948

Experimental Cell Research

Volume 240, Issue 1, 10 April 1998, Pages 66-74

https://doi.org/10.1006/excr.1998.3948 Get rights and content

Abstract

Cellular senescence and limited proliferative capacity of normal diploid cells has a dominant phenotype over immortality of cancerous cells, suggesting its regulation by the expression of a set of genes. In order to isolate the genes that associate with senescence, we have employed a clonal system of conditional SV40 T antigen rat embryo fibroblast cell lines which undergo senescence upon T antigen inactivation. Construction of cDNA libraries from two conditional cell lines and application of differential screening and subtractive hybridization techniques have resulted in the cloning of eight senescence-induced genes (SGP-2/Apo J, α1-procollagen, osteonectin, fibronectin, SM22, cytochrome C oxidase, GTP-α, and a novel gene) and a senescence-repressed gene (FRS-2). Three of these genes encode for extracellular matrix proteins, others are involved in the calcium-dependent signal transduction pathways, while the SGP-2/Apo J gene may have a cellular protective function. RNA analysis has shown that the senescence-associated genes are overexpressed in both normal rat embryonic fibroblasts and human osteoblasts cell cultures undergoing agingin vitro.In comparison, the expression of these genes in a rat fibroblast immortalized cell line (208F cells) was down-regulated after both its partial and its full transformation byrasoncogenes. Thus, cloning of senescence-associated genes opens up new ways to elucidate and/or to modulate aging and cancer.

References (50)

J. Campisi
Cell
(1996)
Y. Satoh et al.
Mutat. Res.
(1994)
A. Noda et al.
Exp. Cell Res.
(1994)
R. Wadhwa et al.
J. Biol. Chem.
(1993)
H. Tahara et al.
Biochem. Biophys. Res. Commun.
(1994)
B. Lecka-Czernik et al.
Exp. Gerontol.
(1996)
M. Martin et al.
Exp. Cell Res.
(1990)
V. Janapati et al.
Mech. Ageing Dev.
(1995)
H.J. Armbrecht et al.
Biochem. Biophys. Acta
(1994)
R. Thweatt et al.
Biochem. Biophys. Res. Commun.
(1992)

H.V. de Silva et al.

J. Biol. Chem.

(1990)

S. Bettuzzi et al.

FEBS Lett.

(1994)

S. Bettuzzi et al.

Biochem. Biophys. Res. Commun.

(1991)

E.W. Khandjian et al.

Exp. Cell Res.

(1992)

E.S. Gonos et al.

Int. J. Oncol.

(1992)

J.R. Smith et al.

Science

(1996)

A. Derventzi et al.

Anticancer Res.

(1996)

S.I.S. Rattan

FASEB J.

(1995)

M.J. Nuell et al.

Mol. Cell Biol.

(1991)

J.A.M. Maier et al.

Science

(1990)

H. Tahara et al.

Oncogene

(1995)

S. Murano et al.

Mol. Cell Biol.

(1991)

M.H.K. Linskens et al.

Nucleic Acids Res.

(1995)

O. Sugawara et al.

Science

(1990)

Cited by (92)

The paradox of FGFR3 signaling in skeletal dysplasia: Why chondrocytes growth arrest while other cells over proliferate
2014, Mutation Research - Reviews in Mutation Research
Somatic mutations in receptor tyrosine kinase FGFR3 cause excessive cell proliferation, leading to cancer or skin overgrowth. Remarkably, the same mutations inhibit chondrocyte proliferation and differentiation in developing bones, resulting in skeletal dysplasias, such as hypochondroplasia, achondroplasia, SADDAN and thanatophoric dysplasia. A similar phenotype is observed in Noonan syndrome, Leopard syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Costello syndrome, Legius syndrome and cardiofaciocutaneous syndrome. Collectively termed RASopathies, the latter syndromes are caused by germline mutations in components of the RAS/ERK MAP kinase signaling pathway. This article considers the evidence suggesting that FGFR3 activation in chondrocytes mimics the activation of major oncogenes signaling via the ERK pathway. Subsequent inhibition of chondrocyte proliferation in FGFR3-related skeletal dysplasias and RASopathies is proposed to result from activation of defense mechanisms that originally evolved to safeguard mammalian organisms against cancer.
Expression profiles of subtracted mRNAs during cellular senescence in human mesenchymal stem cells derived from bone marrow
2013, Experimental Gerontology
Cellular senescence is an irreversible cell cycle arrest that limits the replicative lifespan of cells. Senescence suppresses development of tumors by regulating aging factors, such as cyclin dependent kinase inhibitor (CKI) and telomerase. Suppression subtractive hybridization (SSH) was used to identify genes that were differentially expressed between young human mesenchymal stem cells (Y-hMSCs) and senescent human mesenchymal stem cells (S-hMSCs). We selected positive clones that were functionally characterized by referring to public databases using NCBI BLAST tool. This search revealed that 19 genes were downregulated, and 43 genes were upregulated in S-hMSCs relative to Y-hMSCs. Among subtracted clones in Y-hMSCs, most of genes markedly were related to metabolic functions. These genes, PDIA3, WDR1, FSTL1, COPG1, LMAN1, and PDIA6, significantly downregulated. Conversely, genes for subtracted clones in S-hMSCs were mostly associated with cell adhesion. In particular, the expression levels of 9 genes, HSP90B1, EID1, ATP2B4, DDAH1, PRNP, RAB1A, PGS5, TM4SF1 and SSR3, gradually increased during senescence. These genes have not previously been identified as being related to cellular senescence, but they seemed to be potentially affected during cellular senescence.
Over-expression of human clusterin increases stress resistance and extends lifespan in Drosophila melanogaster
2012, Biochemical and Biophysical Research Communications
Clusterin is a disulfide-linked heterodimeric glycoprotein that has been implicated in a variety of biological processes. Its expression has been shown to be elevated during cellular senescence and normal aging, but it is uncertain whether clusterin protects against aging or whether its expression is a consequence of aging. To investigate the functions of clusterin during organismal aging, we established transgenic Drosophila alleles to induce the expression of the secretory form of human clusterin (hClu^S) using the Gal4/UAS system. hClu^S protein (∼60 kDa) was detected in both adult homogenates and larval hemolymphs of flies ubiquitously overexpressing hClu^S (da-Gal4 > UAS-hClu^S) and in motoneurons (D42-Gal4 > UAS-hClu^S). Interestingly, the mean lifespans of these hClu^S-overexpressing flies were significantly greater than those of control flies that exhibited no hClu^S induction. hClu^S-overexpressing flies also showed significantly greater tolerance to heat shock, wet starvation, and oxidative stress. Furthermore, amounts of reactive oxygen species (ROS) in whole bodies were significantly lower in hClu^S-overexpressing flies. In addition, clusterin was found to prevent the inactivation of glutamine synthetase (GS) by metal-catalyzed oxidation (MCO) in vitro, and this protection was only supported by thiol-reducing equivalents, such as, DTT or GSH, and not by ascorbate (a non-thiol MCO system). Furthermore, this protection against GS inactivation by clusterin was abolished by reacting clusterin with N-ethylmaleimide, a sulfhydryl group-modifying agent. Taken together, these results suggest that a disulfide-linked form of clusterin functions as an antioxidant protein via its cysteine sulfhydryl groups to reduce ROS levels and delay the organismal aging in fruit flies.
At the stem of youth and health
2011, Pharmacology and Therapeutics
Cellular senescence is a specialized form of growth arrest, confined to mitotic cells, induced by various stressful stimuli and characterized by a permanent growth arrest, resistance to apoptosis, an altered pattern of gene expression and the expression of some markers that are characteristic, although not exclusive, to the senescent state. Senescent cells profoundly modify neighboring and remote cells through the production of an altered secretome, eventually leading to inflammation, fibrosis and possibly growth of neoplastic cells. Mammalian aging has been defined as a reduction in the capacity to adequately maintain tissue homeostasis or to repair tissues after injury. Tissue homeostasis and regenerative capacity are nowadays considered to be related to the stem cell pool present in every tissue. For this reason, pathological and patho-physiological conditions characterized by altered tissue homeostasis and impaired regenerative capacity can be viewed as a consequence of the reduction in stem cell number and/or function. Last, cellular senescence is a double-edged sword, since it may inhibit the growth of transformed cells, preventing the occurrence of cancer, while it may facilitate growth of preneoplastic lesions in a paracrine fashion; therefore, interventions targeting this cell response to stress may have a profound impact on many age-related pathologies, ranging from cardiovascular disease to oncology. Aim of this review is to discuss both molecular mechanisms associated with stem cell senescence and interventions that may attenuate or reverse this process.
SM22α-induced activation of p16<sup>INK4a</sup>/retinoblastoma pathway promotes cellular senescence caused by a subclinical dose of γ-radiation and doxorubicin in HepG2 cells
2010, Biochemical and Biophysical Research Communications
Smooth muscle protein 22-alpha (SM22α) is known as a transformation- and shape change-sensitive actin cross-linking protein found in smooth muscle tissue and fibroblasts; however, its functional role remains uncertain. We reported previously that SM22α overexpression confers resistance against anti-cancer drugs or radiation via induction of metallothionein (MT) isozymes in HepG2 cells. In this study, we demonstrate that SM22α overexpression leads cells to a growth arrest state and promotes cellular senescence caused by treatment with a subclinical dose of γ-radiation (0.05 and 0.1 Gy) or doxorubicin (0.01 and 0.05 μg/ml), compared to control cells. Senescence growth arrest is known to be controlled by p53 phosphorylation/p21^WAF1/Cip1 induction or p16^INK4a/retinoblastoma protein (pRB) activation. SM22α overexpression in HepG2 cells elevated p16^INK4a followed by pRB activation, but did not activate the p53/p21^WAF1/Cip1 pathway. Moreover, MT-1G, which is induced by SM22α overexpression, was involved in the activation of the p16^INK4a/pRB pathway, which led to a growth arrest state and promoted cellular senescence caused by damaging agents. Our findings provide the first demonstration that SM22α modulates cellular senescence caused by damaging agents via regulation of the p16^INK4a/pRB pathway in HepG2 cells and that these effects of SM22α are partially mediated by MT-1G.
FGFR3 signaling induces a reversible senescence phenotype in chondrocytes similar to oncogene-induced premature senescence
2010, Bone
Oncogenic activation of the RAS–ERK MAP kinase signaling pathway can lead to uncontrolled proliferation but can also result in apoptosis or premature cellular senescence, both regarded as natural protective barriers to cell immortalization and transformation. In FGFR3-related skeletal dyplasias, oncogenic mutations in the FGFR3 receptor tyrosine kinase cause profound inhibition of cartilage growth resulting in severe dwarfism, although many of the precise mechanisms of FGFR3 action remain unclear. Mutated FGFR3 induces constitutive activation of the ERK pathway in chondrocytes and, remarkably, can also cause both increased proliferation and apoptosis in growing cartilage, depending on the gestational age. Here, we demonstrate that FGFR3 signaling is also capable of inducing premature senescence in chondrocytes, manifested as reversible, ERK-dependent growth arrest accompanied by alteration of cellular shape, loss of the extracellular matrix, upregulation of senescence markers (α-GLUCOSIDASE, FIBRONECTIN, CAVEOLIN 1, LAMIN A, SM22α and TIMP 1), and induction of senescence-associated β-GALACTOSIDASE activity. Our data support a model whereby FGFR3 signaling inhibits cartilage growth via exploiting cellular responses originally designed to eliminate cells harboring activated oncogenes.

View all citing articles on Scopus

¹: To whom correspondence and reprint requests should be addressed at National Hellenic Research Foundation, Institute of Biological Research and Biotechnology, 48 Vas. Constantinou Ave., Athens 11635, Greece. Fax: 30-1-7234008. E-mail:[email protected].

View full text

Regular ArticleCloning and Identification of Genes That Associate with Mammalian Replicative Senescence

Abstract

Cell

Mutat. Res.

Exp. Cell Res.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Exp. Gerontol.

Exp. Cell Res.

Mech. Ageing Dev.

Biochem. Biophys. Acta

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

FEBS Lett.

Biochem. Biophys. Res. Commun.

Exp. Cell Res.

Int. J. Oncol.

Science

Anticancer Res.

FASEB J.

Mol. Cell Biol.

Science

Oncogene

Mol. Cell Biol.

Nucleic Acids Res.

Science

Regular Article
Cloning and Identification of Genes That Associate with Mammalian Replicative Senescence