| Gene name: | CD40L |
| Aging type: | Prevent |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | A549,H460 |
| Gene ID: | 100286596 |
| Category: | protein coding |
| Phenotype: | Lung cancer |
| Experimental category: | L |
| PMID: | 29565449 |
| Experiment: | Western blot//SA-β-gal activity assay//CCK-8 assay//Flow cytometry |
| Description: | We used pcDNA3.1+, pcDNA3.1+-CD40L-WT and pcDNA3.1+-CD40L-M to transfect CD40-positive NSCLC cell lines. CD40L expression was increased in the A549, A549/TR, A549/DDP and H460 cells 48 h after transfection.In addition, the culture cell size was enlarged in the A549/TR, A549/DDP and H460 cells, a trait associated with senescence. These data were confirmed with SA-β-gal staining, yet SA-β-gal staining was not observed in the A549 cells.CCK-8 assay data showed that CD40L-M expression significantly reduced cell proliferation.Flow cytometry data showed that the percentage of S phase cells was decreased and the percentage of G0/G1 phase cells was increased in the CD40L-M-expressed cell group compared with these percentages noted in the controls or negative cells,indicating that CD40L-M expression blocked cell cycle progression. |
| Regulatory pathway: | ATM-CHK2 |
| R-AG-Pathway: | -- |
| Official symbol(s): | ATM-CHEK2 |
| Pathway experiment: | Western blot//SA-β-gal activity assay |
| Pathway description: | We assessed p-Chk2 and p-Chk1 in CD40L-induced senescent A549/TR cells. Data showed that only p-Chk2 was significantly induced 48 h after transfection, indicating that the ATM/Chk2 pathway was activated in response to CD40L-M-induced senescence.We treated CD40L-M-transfected A549/TR cells with an inhibitor of ATM kinase, KU-55933. KU-55933 inhibited phosphorylation of the ATM target protein Chk2 and decreased SA-β-gal activity , suggesting that ATM/Chk2 mediated CD40L-M-induced senescence. |
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