Thermal response in murine L929 cells lacking ?B-crystallin expression and ?B-crystallin expressing L929 transfectants

Blackburn, Robert T.; Galoforo, Sandra; Berns, Christine M.; Ireland, Mark E.; Cho, Joong M.; Corry, Peter M.

doi:10.1007/bf00714333

Cited by 10 publications

(7 citation statements)

References 35 publications

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“…It was reported that overexpression of HSP27 by transfection into rodent and Chinese hamster cell lines directly correlated with survival from hyperthermia [5][6][7] . Overexpression of αB-crystallin has a similar effect [8][9][10] . It was also reported that overexpression of both HSP27 and αB-crystallin protected against ischemic injury in cardiac myocytes [11] .…”

Section: Introductionmentioning

confidence: 92%

Overexpression of heat-shock protein 20 in rat heart myogenic cells confers protection against simulated ischemia/reperfusion injury1

Zhu

Wang

2005

Acta Pharmacologica Sinica

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Aim: To explore whether overexpression of the small heat shock protein HSP20 in rat cardiomyocytes protects against simulated ischemia/reperfusion (SI/R) injury. Methods: Recombinant adenovirus expressing HSP20 was used to infect rat H9c2 cardiomyocytes at high efficiency, as assessed by green fluorescent protein. H9c2 cells were subjected to SI/R stress; survival was estimated through assessment of lactate dehydrogenase and cell apoptosis through caspase‐3 activity. Results: Overexpression of HSP20 decreased lactate dehydrogenase release by 21.5% and caspase‐3 activity by 58.8%. Pretreatment with the protein kinase C inhibitor Ro‐31‐8220 (0.1 μmol/L) for 30 min before SI/R canceled the protective effect of HSP20. The selective mitochondrial K+ATP channel inhibitor 5‐hydroxydecanoate (100 μmol/L) had a similar effect. However, the non‐selective K+ATP channel inhibitor glibenclamide (100 μmol/L) had no significant effect. Conclusion: These data indicate that the protective effect of HSP20 in vitro is primarily due to reduced necrotic and apoptotic death of cardiomyocytes, possibly via the protein kinase C/mitochondrial K+ATP pathway.

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Section: Introductionmentioning

confidence: 92%

Overexpression of heat-shock protein 20 in rat heart myogenic cells confers protection against simulated ischemia/reperfusion injury1

Zhu

Wang

2005

Acta Pharmacologica Sinica

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“…The slight, background fluorescence seen in the nuclei of Figure 1d was attributed to the nonspecific staining by the fluorescent secondary antibody (data not shown). A similar comparison could not be made for L929 cells because they lack endogenous Hsp27 and ␣-b crystalline (Lee et al 1992;Blackburn et al 1996).…”

Section: Characterization Of the Chimeric Hsp27 Proteinmentioning

confidence: 99%

Stress protection by a fluorescent Hsp27 chimera that is independent of nuclear translocation or multimeric dissociation

Borrelli

Bernock²,

Landry³

et al. 2002

Cell Stress Chaper

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A chimeric protein consisting of enhanced green fluorescent protein (EGFP) fused to the N-terminus of human Hsp27 conferred stress protection in human A549 lung carcinoma and murine L929 cells that were stably transfected to express the chimera constitutively. The resultant protection was comparable with that in the same cell lines when they were transfected to express corresponding levels of Hsp27. Unlike L929 cells, A549 cells exhibit endogenous Hsp27 expression, whose expression was inhibited in proportion to the amount of fluorescent chimera expressed, suggesting that the A549 cells recognized the latter as Hsp27. Upregulation of Hsp27 or chimeric Hsp27 in all transfected cell lines (stable or transient transfection) caused no measurable change in cellular glutathione levels, indicating that glutathione played no role in the stress protection associated with either protein. Chimeric Hsp27 had a monomeric molecular weight of 55 kDa (that of Hsp27 plus EGFP) in both cell types and formed a 16-mer complex twice as massive as that formed by Hsp27. Heat shock or sodium arsenite induced phosphorylation of both chimeric Hsp27 and Hsp27, which resulted in the disaggregation of Hsp27 multimers in both cell types and disaggregation of 20% of the chimeric multimers in L929 cells. But chimeric Hsp27 multimers did not disaggregate after stress in A549 cells. Epifluorescence and confocal microscopy demonstrated that chimeric Hsp27 was restricted to the cytoplasm under normal growth conditions and after heat shock in all cells. This study supports the conclusions that Hsp27 stress protection requires neither its translocation into the nucleus nor the dissociation of its multimeric complex. Furthermore, it demonstrates that fluorescent chimeras of heat shock proteins can be functional and used to observe the protein's distribution within living cells.

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“…To further examine the role of HSP27 in growth, actin polymerization, and cell protection against stress, we transfected murine fibrosarcoma L929 cells (devoid of constitutively expressed small HSPs [Lee et al, 1992;Blackburn et al, 1996]) and human osteoblastlike SaOS-2 cells with the constitutive expression vector pSVL carrying the coding sequence of the human hsp27 gene. We examined the resulting HSP27 expression and the associated effects on growth, morphology, actin filament organization as well as on the cell sensitivity to heat shock and to different cytotoxic agents.…”

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confidence: 99%

Expression of HSP27 results in increased sensitivity to tumor necrosis factor, etoposide, and H2O2 in an oxidative stress-resistant cell line

Mairesse¹,

Bernaert²,

Bino³

et al. 1998

J. Cell. Phys.

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The role of HSP27 in cell growth and resistance to stress was investigated using murine fibrosarcoma L929 cells (normally devoid of constitutively expressed small HSPs) and human osteoblast-like SaOS-2 cells stably transfected with a human hsp27 expression vector. Our data showed that our L929 cells were more resistant to oxidative stress than generally observed for this line. Production of HSP27 in these cells led to a marked decrease in growth rate associated with a series of phenotypical changes, including cell spreading, cellular and nuclear hypertrophy, development of an irregular outline, and a tremendous accumulation of actin stress fibers. By contrast, none of these changes was observable in SaOS-2/hsp27 transfectants overexpressing the protein product. Together, these observations are consistent with a cause-to-effect cascade relationship between increased (or induced) HSP27 expression, changes in cytoskeletal organization, and decreased growth. On the other hand, whereas the transfection of the hsp27 gene increased the cell resistance to heat in both cell lines, only in SaOS-2 cells was this associated with protection to the cytotoxic action of tumor necrosis factor-alpha (TNF-alpha) and etoposide. Unexpectedly, L929/hsp27 transfectants exhibited an increased sensitivity to both agents and also to H2O2. These data thus imply that different mechanisms are involved in the cell resistance to heat shock and to the cytotoxic action of TNF-alpha, etoposide, and H2O2. They also plead against the simple view that overexpression of a phosphorylatable HSP27 would necessarily be beneficial in terms of increased cell resistance to any type of stress. Our data further indicate that the role of HSP27 in cellular resistance to stress and in cell proliferation involves different targets and that the ultimate result of its interference with these processes depends on the intracellular context in which the protein is expressed.

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Thermal response in murine L929 cells lacking ?B-crystallin expression and ?B-crystallin expressing L929 transfectants

Cited by 10 publications

References 35 publications

Overexpression of heat-shock protein 20 in rat heart myogenic cells confers protection against simulated ischemia/reperfusion injury1

Overexpression of heat-shock protein 20 in rat heart myogenic cells confers protection against simulated ischemia/reperfusion injury1

Stress protection by a fluorescent Hsp27 chimera that is independent of nuclear translocation or multimeric dissociation

Expression of HSP27 results in increased sensitivity to tumor necrosis factor, etoposide, and H2O2 in an oxidative stress-resistant cell line

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