| External factors: | BBR |
| Aging type: | Accelerate |
| Aging characteristic: |
| Category: | Chemical compounds |
| Phenotype: | Bladder cancer |
| Experiment: | SA-β-gal activity assay//MTT assay//Colony formation assay |
| Description: | The results showed that BBR treatment significantly increased the number of SA-β-gal-positive cells, indicating that BBR induces cell senescence in 253J cells.MTT assays showed that BBR significantly decreased cell viability in a dose-dependent manner. The results of the colony formation assay showed that BBR induced a concentration-dependent decrease in the number of colonies.BBR treatment increased the proportion of cells in the G0/G1 phase, accompanied by a decrease in the proportion of cells in the S and G2/M phases in T24 cells. BBR increased the proportion of cells in the S phase in 5637 cells, along with a decline in the proportion of that in G0/G1 and G2/M phases. In 253J cells, BBR caused an increase in the proportion of cells in the G2/M phase.A western blot analysis of the cell cycle-associated proteins revealed that the protein levels of CDK2, CDK1, cyclin A, and cyclin D1 were significantly decreased in BBR-treated BCa cells. |
| Target gene: | MIR-17-5P |
| R-EF-Target gene: | Upregulation |
| Official symbol(s): | MIR-17-5P |
| Target gene experiment: | qRT-PCR |
| Target gene description: | qRT-PCR assays showed that among these five miRNAs, only miR-17-5p level was upregulated by BBR. Further analysis revealed that BBR promoted the expression of miR-17-5p in a concentration-dependent manner. |
| Regulatory pathway: | JAK1-STAT3 |
| R-EF-Pathway: | Downregulation |
| Pathway experiment: | qPCR//Western blot//SA-β-gal activity assay//Dual-Luciferase reporter assay |
| Pathway description: | BBR treatment did not decrease the mRNA levels of JAK1 and STAT3. Moreover, the decreased protein levels of JAK1 and STAT3 caused by BBR treatment could not be restored by MG132 treatment. The results of western blot and IHC ana- lyses of xenograft tumors confirmed that BBR decreased JAK1, STAT3, and p-STAT3 protein levels in vivo.BBR significantly decreased the protein levels of JAK1 and JAK2. Both nuclear STAT3 and p-STAT3 protein levels were downregulated by BBR treatment.Luciferase analysis confirmed an obvious decrease in the transcriptional activity of STAT3 in BBR-treated cells. |
Annotation: