CAJ | 학술논문

目的：在3T3-L1细胞模型中，探讨组蛋白乙酰转移酶PCAF对脂肪细胞分化调控的分子机制。方法：应用Real-time PCR和Western blot检测PCAF在3T3-L1细胞分化过程中的表达；应用重组慢病毒感染细胞，shRNA干扰或过表达PCAF，通过Oil-Red-O染色法观察PCAF表达量变化对细胞分化的影响；应用ChIP on chip和PCR array的方法筛选PCAF调控的下游基因和通路；应用Western blot和ChIP qPCR对潜在目的基因进行验证。结果：PCAF在3T3-L1细胞分化过程中表达升高；PCAF表达下调会抑制脂肪细胞的分化；PCAF可以在细胞分化不同时间点诱导PDPK1和FOXO1的表达，并间接影响了FOXO1磷酸化。结论：组蛋白乙酰转移酶PCAF通过胰岛素通路（PI3K/PDPK1/AKT/FOXO1）参与对脂肪细胞分化的转录调控。
목적：재3T3-L1세포모형중，탐토조단백을선전이매PCAF대지방세포분화조공적분자궤제。방법：응용Real-time PCR화Western blot검측PCAF재3T3-L1세포분화과정중적표체；응용중조만병독감염세포，shRNA간우혹과표체PCAF，통과Oil-Red-O염색법관찰PCAF표체량변화대세포분화적영향；응용ChIP on chip화PCR array적방법사선PCAF조공적하유기인화통로；응용Western blot화ChIP qPCR대잠재목적기인진행험증。결과：PCAF재3T3-L1세포분화과정중표체승고；PCAF표체하조회억제지방세포적분화；PCAF가이재세포분화불동시간점유도PDPK1화FOXO1적표체，병간접영향료FOXO1린산화。결론：조단백을선전이매PCAF통과이도소통로（PI3K/PDPK1/AKT/FOXO1）삼여대지방세포분화적전록조공。
O bjective:To investigate the molecular mechanism of P300/CBP associated factor (PCAF) gene in regulation of adipocytes differentiation. Methods: PCAF expression levels were measured by Real-time PCR and Western blot in 3T3-L1 cells during differentiation;3T3-L1 cells were transfected by recombinant Lentiviral ORF/shRNA and the effects of PCAF overexpression or knockdown on differentiation were observed by Oil-Red-O staining;ChIP on chip and PCR array were used to screen the signaling pathways regulated by PCAF;expressions of potential target genes and pathways were detected by Western blot and ChIP qPCR. Results:PCAF expression increased during 3T3-L1 cells differentiation and adipogenesis;the differentiation of 3T3-L1 might be interfered by knockdown of PCAF;expression of PDPK1 and FOXO1 were induced by PCAF at various time points during 3T3-L1 differentiation; the phosphorylation of FOXO1 was also regulated by PCAF indirectly. Conclusion: PCAF regulates preadipocytes differentiation through the insulin pathway (PI3K/PDPK1/AKT/FOXO1).