AKT不同亚型干扰对成人脂肪干细胞成脂分化的影响  

Effect of Different Subtypes of AKT Interference on Adipogenesis of Human Adipose-derived Stem Cells

作　　者：李付贵[1,2] 陈京[1] 程伟民[1] 季明芳[1] 

机构地区：[1]中山大学肿瘤防治中心,广东广州510060 [2]中山大学附属中山市人民医院肿瘤研究所,广东中山528400

出　　处：《中山大学学报（医学科学版）》2015年第5期676-682,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金博士启动基金(2014A030310013);中国博士后基金(2014M562244);中山市科技局面上项目(2014A1FC047)

摘　　要：【目的】探讨AKT不同亚型AKT1和AKT2在成人脂肪干细胞成脂分化过程的表达及意义。【方法】取生长状态良好的P3代的成人脂肪组织来源的间充质干细胞(h ADSC),以负载AKT1-sh RNA和AKT2-SHRNA的慢病毒载体分别转染h ADSC,获得稳定干扰AKT1和AKT2的细胞株(分别设为AKT1-3组和AKT2-1组),转染非干扰序列NTC-sh RNA的设为对照组。将上述细胞分别进行成脂肪诱导,观察脂滴的形成,并检测相关成脂肪基因的表达和成脂肪关键转录因子PPARγ的表达。【结果】首先成功获得稳定表达AKT1-sh RNA和AKT2-sh RNA的h ADSC。与对照组细胞的成脂过程相比,AKT1-3组h ADSC成脂分化并无明显影响;而AKT2-1组脂滴形成明显减少,油红O染色与定量也显示相应改变(P<0.01)。成脂相关基因定量结果也发现,FABP4、LPL及GPDH等表达明显下降(P<0.01)。此外,AKT1-3组诱导后下游转录因子PPARγ的表达与NTC组相比,未见明显改变;而AKT2-1组的PPARγ表达明显下降,尤其是PPARγ2的降低更为明显。【结论】AKT2干扰导致h ADSC成脂肪过程严重抑制,而AKT1干扰则并不能影响该过程。【Objective】 To explore the role of different subtypes of AKT, AKT1 and AKT2 interference in the differentiation of human adipose-derived stem cells. 【Methods】 Human adipose-derived stem cell（h ADSC） of third generation with good state were transfected with lentiviral vectors carrying AKT1 sh RNA or AKT2 sh RNA respectively, to obtain stable cell lines expressing AKT1 or AKT2-sh RNA（set as AKT1-3 group and AKT2-1 group）, and h ADSC transfected with NTC-sh RNA were set as control group. After the above-mentioned cell lines were induced into adipocytes, we observed the formation of lipid droplets, and detected the m RNA expression of adipocyte-related genes and key transcription factors of adipogenesis, PPAR gamma expression. 【Results】 h ADSC carrying AKT1-sh RNA or AKT2-sh RNA was successfully obtained. Compared with control group, AKT1-3 group had no obvious changes in the lipid-droplet formation, while AKT2-1 group almost had no lipid-droplet formation. Oil red O staining and quantitative analysis also showed the corresponding changes（P 〈0.01）. Adipocyte-related gene quantitative expression results also manifested that FABP4, LPL and GPDH expression decreased obviously in AKT2-1 group（P〈 0.01）. Additionally, downstream transcription factor PPAR gamma expression was significantly inhibited in AKT2-1 group compared with NTC group, however no significant changes were observed in AKT1-3 group. 【Conclusion】 AKT2 played a critical role in adipogenesis of h ADSC, whereas AKT1 did not significantly affected the process.

关 键 词：脂肪干细胞 AKT 成脂肪分化 sh RNA 

分 类 号：R723.14[医药卫生—儿科]