论文部分内容阅读
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PCRarray的方法筛选PCAF调控的下游基因和通路;应用Westernblot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。
OBJECTIVE: To investigate the molecular mechanism of histone acetyltransferase (PCAF) regulation of adipocyte differentiation in 3T3-L1 cell model. Methods: The expression of PCAF in 3T3-L1 cells was detected by Real-time PCR and Western blot. The expression of PCAF was detected by Oil-Red-O staining after infected with recombinant lentivirus, shRNA interference or overexpression of PCAF. On the differentiation of cells. The downstream genes and pathways regulated by PCAF were screened by ChIP on chip and PCRarray. The potential target genes were verified by Western blot and ChIP qPCR. Results: The expression of PCAF was increased during the differentiation of 3T3-L1 cells. Down-regulation of PCAF inhibited the differentiation of adipocytes. PCAF induced the expression of FOXO1 and FOXO1 at different time points, and indirectly affected FOXO1 phosphorylation. CONCLUSION: Histone acetyltransferase PCAF is involved in the transcriptional regulation of adipocyte differentiation via the insulin pathway (PI3K / PDPK1 / AKT / FOXO1).