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目的探讨miR-17对人包皮成纤维细胞(HFF)衰老的影响。方法 miR-17重组慢病毒感染细胞,qRT-PCR确定感染效率,CCK-8法观察miR-17对细胞增殖的影响,衰老相关β-半乳糖苷酶染色观察miR-17对细胞衰老的影响,流式细胞法检测miR-17对细胞周期的影响,Western blotting检测cyclin D1和p21蛋白的表达。结果成功分离得到HFF,并建立了稳定表达miR-17的HFF-miR-17细胞系。且HFF-miR-17细胞增殖能力明显升高,β-半乳糖苷酶染色阳性率降低;S期细胞群比例明显增高,细胞周期蛋白cyclin D1表达显著上调,p21蛋白明显下调。结论 miR-17可通过上调cyclin D1蛋白、下调p21蛋白促进原代细胞的增殖,抑制原代细胞的衰老。
Objective To investigate the effect of miR-17 on the aging of human foreskin fibroblasts (HFF). Methods The infected cells were infected with miR-17 recombinant lentivirus and the infection efficiency was determined by qRT-PCR. The effect of miR-17 on cell proliferation was observed by CCK-8 assay. The effect of miR-17 on cell senescence was observed by aging- The effect of miR-17 on cell cycle was detected by flow cytometry. The expression of cyclin D1 and p21 protein was detected by Western blotting. Results HFF was successfully isolated and the HFF-miR-17 cell line stably expressing miR-17 was established. The proliferation of HFF-miR-17 cells was significantly increased and the positive rate of β-galactosidase staining was decreased. The proportion of S phase cells was significantly increased, cyclin D1 expression was significantly increased, and p21 protein was significantly down-regulated. Conclusion miR-17 can up-regulate cyclin D1 protein and down-regulate p21 protein to promote the proliferation of primary cells and inhibit the senescence of primary cells.