| Gene name: | RAD21 |
| Aging type: | Prevent |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | MDA-MB-231,HCT116,LNCaP,DU 145 |
| Experiment: | BrdU assay//SA-β-gal activity assay//Immunofluorescence |
| Description: | The more effective siRNA #2 was used in the subsequent experiments, which showed that RAD21 suppression significantly inhibited MDA-MB-231 cell proliferation , decreased bromodeoxyuridine (BrdU) incorporation, and elevated the activity of the senescence marker SA-β-gal.Heterochromatin foci are a typical signature of human cells undergoing senescence.Heterochromatin foci were readily visible in senescent cells transfected with RAD21 siRNA after DAPI staining, with 45% of nuclei displaying a punctate DAPI staining pattern.We also found that RAD21 silencing induced senescence in HCT116 colon cancer cells, as well as LNCap and DU145 prostate cancer cells . |
| Target gene: | C-MYC |
| Official symbol(s): | C-MYC |
| R-AG-Target gene: | -- |
| Subcategory: | Unclear |
| Target gene experiment: | Western blot//SA-β-gal activity assay |
| Target gene description: | c-Myc overexpression in cells transfected with RAD21 siRNA and the pcDNA3.1-c-Myc expression plasmid did not result in reduction of c-Myc protein, whereas decrease in c-Myc protein was induced in cells cotransfected with RAD21 siRNA and the control plasmid. Furthermore, c-Myc overexpression could also lead to functional inactivation of RB1 by increasing its phosphorylation in cells transfected with RAD21 siRNA. As expected,co-expression of RAD21 siRNA and c-Myc protein dramatically reduced the number of senescent cells. |
| Regulatory pathway: | RB-E2F |
| R-AG-Pathway: | -- |
| Official symbol(s): | RB1-E2F1 |
| Pathway experiment: | SA-β-gal activity assay//Western blot |
| Pathway description: | RAD21-depleted MDA-MB-231 cells displayed lower levels of phospho-RB1 .all phenotypic markers of cellular senescence tested were significantly inhibited when either p21 or RB1 was silenced with RAD21.RAD21 siRNA-induced senescence was characterized by RB1 hypophosphorylation, downregulation of the E2F-dependent genes proliferating cell nuclear antigen (PCNA), cyclinA, and cyclinD1, as well as downregulation of c-Myc, a key regulator of the RB1/E2F pathway.c-Myc overexpression could also lead to functional inactivation of RB1 by increasing its phosphorylation in cells transfected with RAD21 siRNA. |
Annotation:
Loading,please wait...