Glucocorticoids Induce G1 as Well as S-Phase Lengthening in Normal Human Stimulated Lymphocytes: Differential Effects on Cell Cycle Regulatory Proteins

Baghdassarian, Nathalie; Catallo, Régine; Mahly, Mohamed A.; Ffrench, Patrick; Chizat, Florence; Bryon, Paul‐André; Ffrench, Martine

doi:10.1006/excr.1998.3942

Cited by 35 publications

(32 citation statements)

References 27 publications

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“…Harvested cells were fixed in 75% ethanol at 4 o C. Cells were then treated with RNase III and stained with propidium iodide as previously described. 50 Cell cycle distribution was analyzed using a FACS Canto II (Becton Dickinson).…”

Section: Methodsmentioning

confidence: 99%

“…BrdU labeling of S-phase cells was performed as previously described. 50,51 p16 and BrdU double labeling was performed by doing first p16 labeling followed by BrdU labeling.…”

Section: Methodsmentioning

confidence: 99%

“…51 and was performed as previously described. 50 Briefly, iododeoxyuridine (IdU, 10 µM, sigma) was added to each cell sample in culture. After 2 h of incubation, cells were rinsed in PBS and then incubated with 10 µM BrdU for 15 min.…”

Section: Methodsmentioning

confidence: 99%

“…Western blot was carried out as previously described. 50 The following primary antibodies used were: mouse monoclonal anti-p16…”

Section: Notementioning

confidence: 99%

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S-phase lengthening induced by p16^INK4aoverexpression in malignant cells with wild-type pRb and p53

et al. 2010

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Section: Methodsmentioning

confidence: 99%

“…BrdU labeling of S-phase cells was performed as previously described. 50,51 p16 and BrdU double labeling was performed by doing first p16 labeling followed by BrdU labeling.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Western blot was carried out as previously described. 50 The following primary antibodies used were: mouse monoclonal anti-p16…”

Section: Notementioning

confidence: 99%

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S-phase lengthening induced by p16^INK4aoverexpression in malignant cells with wild-type pRb and p53

et al. 2010

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“…Loading equality was assessed by Ponceau red staining analysis and quantification of b-actin or a non-specific protein band (Wang et al, 2005;Kaddar et al, 2009). Quantifications were carried out by densitometric analysis using Kodak 1D Image Analysis Software (Sigma) as previously described (Baghdassarian et al, 1998).…”

Section: Western Blotmentioning

confidence: 99%

Cyclin-dependent kinase 1 expression is inhibited by p16INK4a at the post-transcriptional level through the microRNA pathway

Domenech

Catallo

et al. 2010

Oncogene

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The p16INK4a protein regulates cell cycle progression mainly by inhibiting the activity of G1-phase cyclin-dependent kinases (CDKs) 4 and 6, the subsequent retinoblastoma protein (pRb) phosphorylation and E2F transcription factor release. The p16INK4a protein can also repress the activity of other transcription factors, such as c-myc, nuclear factorkappaB and c-Jun/AP1. Here, we report that, in two p16WT and p53 WT cell lines (MCF7 and U87), p16INK4a overexpression induces a dramatic decrease in CDK1 protein expression. In response to p16 INK4a, the decreased rate of CDK1 protein synthesis, its unchanged protein half-life, unreduced CDK1 mRNA steady-state levels and mRNA half-life allow us to hypothesize that p16INK4a could regulate CDK1 expression at the post-transcriptional level. This CDK1 downregulation is mediated by the 3 0 -untranslated region (3 0 UTR) of CDK1 mRNA as shown by translational inhibition in luciferase assays and is associated with a modified expression balance of microRNAs (miRNAs) that potentially regulate CDK1, analyzed by TaqMan Human microRNA Array. The p16 INK4a-induced expression of two miRNAs (miR-410 and miR-650 chosen as an example) in MCF7 cells is confirmed by individual reverse transcriptionqPCR. Furthermore, we show the interaction of miR-410 or miR-650 with CDK1-3 0 UTR by luciferase assays. Endogenous CDK1 expression decreases upon both miRNA overexpression and increases with their simultaneous inhibition. The induction of miR-410, but not miR-650 could be related to the pRb/E2F pathway. These results demonstrate the post-transcriptional inhibition of CDK1 by p16 INK4a . We suggest that p16INK4a may regulate gene expression by modifying the functional equilibrium of transcription factors and consequently the expression balance of miRNAs.

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T-cell Cycle and Immunosenescence: Role of Aging in the T-cell Proliferative Behaviour and Status Quo Maintenance

Witkowski

Handbook on Immunosenescence

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Glucocorticoids Induce G1 as Well as S-Phase Lengthening in Normal Human Stimulated Lymphocytes: Differential Effects on Cell Cycle Regulatory Proteins

Cited by 35 publications

References 27 publications

S-phase lengthening induced by p16^INK4aoverexpression in malignant cells with wild-type pRb and p53

S-phase lengthening induced by p16^INK4aoverexpression in malignant cells with wild-type pRb and p53

Cyclin-dependent kinase 1 expression is inhibited by p16INK4a at the post-transcriptional level through the microRNA pathway

T-cell Cycle and Immunosenescence: Role of Aging in the T-cell Proliferative Behaviour and Status Quo Maintenance

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Glucocorticoids Induce G1 as Well as S-Phase Lengthening in Normal Human Stimulated Lymphocytes: Differential Effects on Cell Cycle Regulatory Proteins

Cited by 35 publications

References 27 publications

S-phase lengthening induced by p16INK4aoverexpression in malignant cells with wild-type pRb and p53

S-phase lengthening induced by p16INK4aoverexpression in malignant cells with wild-type pRb and p53

Cyclin-dependent kinase 1 expression is inhibited by p16INK4a at the post-transcriptional level through the microRNA pathway

T-cell Cycle and Immunosenescence: Role of Aging in the T-cell Proliferative Behaviour and Status Quo Maintenance

Contact Info

Product

Resources

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S-phase lengthening induced by p16^INK4aoverexpression in malignant cells with wild-type pRb and p53

S-phase lengthening induced by p16^INK4aoverexpression in malignant cells with wild-type pRb and p53