External factors
| External factors: |
Low frequency magnetic fields |
| 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. |
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Regulatory relationship
| 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. |
| Official symbol(s): |
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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. |
Aging network
Annotation:
The green line represents Upregulation.
The purple line represents Downregulation.
The orange line represents Activation.
The yellow line represents Inhibition.
The gray line represents Unclear.
Pathway view
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