| Gene name: | MIR22 |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | MRC-5,Fibroblast |
| Experiment: | qRT-PCR//Western blot//DAPI staining//BrdU assay//Cell proliferation assay//Flow cytometry |
| Description: | Quantitative (q) RT-PCR analysis confirmed that miR-22 expression was increased in senescent TIG-3 and other fibroblasts and even up-regulated by more than fivefold in senescent MRC-5 cells.miR-22 also increased SA-β-gal activity miR-22 overexpression induced cell cycle arrest at G1 phase, accompanied by the decrease in percentages of S phase. The inhibitory effect of miR-22 on cell cycle progression has also been recently reported in other cancer cells. |
| Target gene: | SIRT1//SP1//CDK6 |
| Official symbol(s): | SIRT1//SP1//CDK6 |
| R-AG-Target gene: | --// --// -- |
| Subcategory: | Unclear |
| Target gene experiment: | MiRanda,TargetScan,and PicTar//Luciferase reporter assay//Western blot//SA-β-gal activity assay |
| Target gene description: | The mRNAs of SIRT1 and Sp1 contain putative binding sites for miR-22 in their 3-UTRs, and each site is broadly conservative among mammals. CDK6 mRNA contains three miR-22 binding sites in the 3'-UTR, whereas it is different in conservation for each site.In SiHa and MDA-D3 cells, miR-22 significantly reduced the luciferase activities of the full-length 3'-UTR or WT SIRT1, SP1, and CDK6 site 1 and site 3) reporters, compared with the negative cont miR. Western blot analysis showed that overexpression of miR-22 markedly down-regulated SIRT1, SP1, and CDK6 in SiHa and MDA-D3 cells.We forced SiHa or MDA-D3 cells to express SIRT1, Sp1, or CDK6 using plasmid constructs lacking 3-UTRs of these genes. Indeed, miR-22–induced cell growth repression and SA-β-gal activity were partially rescued by the introduction of SIRT1, CDK6, or Sp1 in either SiHa cells or MDA-D3 cells (not depicted), although it seemed weak for the effect of Sp1 overexpression on cell growth, which might be because of the indirect role of Sp1 in the pRb pathway of senescence. |
| Regulatory pathway: | -- |
| R-AG-Pathway: | -- |
| Pathway experiment: | -- |
| Pathway description: | -- |
Annotation:
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