CAJ | 학술논문

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。
목적：탐토 miR-17대인포피성섬유세포（HFF）쇠로적영향。방법 miR-17중조만병독감염세포， qRT-PCR학정감염효솔，CCK-8법관찰 miR-17대세포증식적영향，쇠로상관β-반유당감매염색관찰 miR-17대세포쇠로적영향，류식세포법검측 miR-17대세포주기적영향，Western blotting 검측 cyclin D1화 p21단백적표체。결과성공분리득도 HFF，병건립료은정표체 miR-17적 HFF-miR-17세포계。차 HFF-miR-17세포증식능력명현승고，β-반유당감매염색양성솔강저；S 기세포군비례명현증고，세포주기단백 cyclin D1표체현저상조，p21단백명현하조。결론miR-17가통과상조 cyclin D1단백、하조 p21단백촉진원대세포적증식，억제원대세포적쇠로。
Objective To study the effect of miR-17 on the senescence of human foreskin fibroblasts (HFF).Methods Recombinant lentivirus-expressed miR-17 infected HFF and miR-17 expression was examined with qRT-PCR.The proliferation and senescence of HFF were detected with CCK-8 and SA-β-Gal kit,respectively.Cell cycle profile was observed with flow cytometry.The protein levels of cyclin D1 and p21 were detected with Western blotting.Results HFF were successfully isolated.Compared with HFF-NC,the proliferation ability of HFF-miR-17 cells was obviously increased while the expression of SA-β-Gal in senescent cells decreased.S phase cells of HFF-miR-17 increased signifi-cantly and more HFF-NC cells arrested in the G1 phase.Expression of cyclin D1 was significantly up-regulated in HFF-miR-17,while p21 expression was attenuated.Conclusion miR-17 promotes the proliferation and inhibits senescence of primary cells by upregulating cyclin D1 and downregulating p21 proteins.