External factors: | Low frequency magnetic fields |
Aging type: | Accelerate |
Aging characteristic: |
Category: | Other |
Phenotype: | Lung cancer |
Experiment: | SA-β-gal activity assay//Flow cytometry//CCK-8 assay |
Description: | Flow cytometry//CCK-8 assay:In consistent with previous data, the proliferation of A549 cells and LLC cells were inhibited by exposure to LF-MF. Cell cycle progression is an essential process by which cell monitors its growth and differentiation. Compared to Sham MF, A549 cells exposed to LF-MF displayed strikingly decreased number of cells in the S and G2/M phases and increased arrest in G0/G1 phase from 53.88% to 77.90%.SA-β-gal activity assay: Staining of senescence-associated β-galactosidase (SA-β-gal) in lung cancer cells showed that LF-MF treatment up-regulate frequency of β-galactosidase (β-Gal)-positive cells. |
Target gene: | MIR-34A//P53 |
R-EF-Target gene: | Upregulation//Upregulation |
Official symbol(s): | MIR-34A//P53 |
Target gene experiment: | qRT-PCR//Flow cytometry//SA-β-gal activity assay//Western blot |
Target gene description: | qRT-PCR:Upon checking several famous miRNAs related to cell proliferation, cell cycle and cell senescence, we found miR-34a level was up-regulated to 4.64 folds after 2 Days exposure to LF-MF.Flow cytometry//SA-β-gal activity assay:Here, LLC and A549 cells were transiently transfected with synthetic miR-34a precursor (pre-miR-34a), or antisense miR-34a inhibitor (anti-miR-34a). Compared with negative control, ectopic expression of miR-34a significantly inhibited the proliferation and induced cell G1 phase arrest in A549 cells and LLC cells .Moreover, the transfected A549 and LLC cells with pre-miR-34a appeared a senescence-like phenotype, enlarged cellular size and increased numbers of SA-β-gal positive cells . However, LF-MF treatment could up-regulate protein level of P53 in LLC cells. |
Regulatory pathway: | E2F |
R-EF-Pathway: | -- |
Official symbol(s): | E2F |
Pathway experiment: | qPCR//Western blot |
Pathway description: | Indeed, ectopic expression of miR-34a downregulates the expressions of E2F3 and E2F1 at the translational level and protein level . Importantly, LF-MF exposure could down-regulated both E2F1 and E2F3 protein level in LLC cells. |
Annotation: