RNA interference of PPARγ using fiber-modified adenovirus vector efficiently suppresses preadipocyte-to-adipocyte differentiation in 3T3-L1 cells
RNA interference of PPARγ using fiber-modified adenovirus vector efficiently suppresses preadipocyte-to-adipocyte differentiation in 3T3-L1 cells
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DOI：
10.1016/j.gene.2005.01.005
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发表时间：
2005-03-28
期刊：
影响因子：
3.5
通讯作者：
Hayakawa, T
中科院分区：
文献类型：
--
作者：
Hosono, T;Mizuguchi, H;Hayakawa, T
过氧化物酶体增殖物激活受体（过氧化物酶体增殖物激活受体）γ被认为是脂肪细胞分化的“主调节因子”，并在脂肪组织中大量表达。为了了解脂肪组织中PPAR γ的生物学作用，针对PPAR γ的RNA干扰（RNAi）应该是一个强有力的工具。3 T3-L1细胞系是研究前脂肪细胞向脂肪细胞分化机制的理想模型。然而，该细胞系难以通过质粒载体和病毒载体转染。我们通过纤维修饰的腺病毒（Ad）载体优化了3 T3-L1前脂肪细胞和脂肪细胞的转导。在测试的各种载体中，纤维球C-末端的多聚赖氨酸修饰最显著地提高了3 T3-L1前脂肪细胞和脂肪细胞中的转导效率。然后，我们研究了纤维修饰的Ad载体与多聚赖氨酸肽表达的小干扰RNA（siRNA）的PPAR γ抑制3 T3-L1前脂肪细胞分化成脂肪细胞。油红0染色和甘油-3-磷酸脱氢酶（GPDH）活性测定表明，载体有效地抑制了3 T3-L1前脂肪细胞向脂肪细胞的分化。这些结果表明，含有聚赖氨酸肽的纤维修饰的Ad载体和RNAi的组合是使用3 T3-L1模型研究肥胖症和糖尿病中脂肪形成的生物学和生理学机制的有效工具。Ad载体介导的针对PPAR γ的RNAi也应该可用于阐明除脂肪外的各种组织中的PPAR γ途径的生物学作用，以及用于治疗性应用于各种疾病，包括肥胖症和糖尿病。(c)2005 Elsevier B. V.保留所有权利。
The peroxisome proliferator-activated receptor (PPAR) gamma is regarded as a "master regulator" of adipocyte differentiation and is abundantly expressed in adipose. To understand the biological role of PPAR gamma in adipose, RNA interference (RNAi) of PPAR gamma should be a powerful tool. 3T3-L1 cell line serves an excellent model to investigate the mechanism of preadipocyte-to-adipocyte differentiation. However, this cell line is difficult to transfect by plasmid vectors and viral vectors. We optimized the transduction of both 3T3-L1 preadipocytes and adipocytes by means of fiber-modified adenovirus (Ad) vectors. Among the various vectors tested, polylysine modification of the C-terminal of the fiber knob most markedly improved the transduction efficiency in both 3T3-L1 preadipocytes and adipocytes. Then, we examined whether fiber-modified Ad vectors with polylysine peptides expressing the small interfering RNA (siRNA) for PPAR gamma inhibit the differentiation of 3T3-L1 preadipocytes into adipocytes. Oil red 0 staining and measurement of glycerol-3-phosphate dehydrogenase (GPDH) activity indicated that the vectors effectively suppressed the differentiation of 3T3-L1 preadipocytes to adipocytes. These results suggested that the combination of fiber-modified Ad vectors containing polylysine peptides and RNAi is an effective tool for the study of the biological and physiological mechanism of adipogenesis in adiposity and diabetes using 3T3-L1 models. Ad vector-mediated RNAi for PPAR gamma should also be useful to clarify the biological role of the PPAR gamma pathway in various tissues in addition to adipose and for therapeutic application to a variety of diseases, including adiposity and diabetes. (c) 2005 Elsevier B.V. All rights reserved.