表达谱基因芯片筛选前体脂肪细胞分化相关基因的研究.docxVIP

表达谱基因芯片筛选前体脂肪细胞分化相关基因的研究 【摘要】 目的:运用基因芯片技术研究前体脂肪细胞分化相关基因表达谱的变化。方法: 体外培养前体脂肪细胞，诱导分化为成熟脂肪细胞。提取脂肪细胞总RNA , 纯化mRNA 后逆转录制备目标序列, 应用含有4503种小鼠基因cDNA 的表达谱芯片Mouse430_2进行差异表达谱分析。结果: 在前体脂肪细胞分化的基因表达谱中差异表达基因共有640 条,其中表达水平显著变化的有38条，上调基因2条,下调基因1条。差异表达基因中有细胞凋亡相关基因、原癌基因和抑癌基因、细胞信号和传递蛋白基因等多种基因。这些基因可能与脂肪细胞分化相关疾病的发病机制有关。结论:采用基因芯片技术筛选脂肪细胞分化相关基因,可能为脂肪细胞分化相关疾病病因和发病机制的研究提供新思路。 中国编辑。  【关键词】 前体脂肪细胞；基因表达；基因芯片  [Abstract] Objective: To screen the differently expressed genes during cell differentiated of adipocyteT3-L1 using cDNA microarray. Methods:T3-L1 cells were cultured, induce them differentiated, isolate total RNA. Reversely transcripted to cDNAs with fluorescent dUTP to prepare the target sequences , then the target sequences were hybridized to cDNA microarray of503human genes. After washing , the cDNA microarray were scanned for the fluorescent signals and showed differences between kinds of cells. Results: Among503geneprobes , the expression level of40 genes changed,genes showed dramatically differential expressed profile withupregulated and 1downregulated genes. The differential expressed genes were related with apoptosis , proto or antioncogene , cell signal conduct , and ytoskeleton , etc. These genes might be related with the pathogenesis of adipocyte differentiation. Conclusion: The investigations based on cDNA microarray can sequence related genes in adipocyte differentiation , and may provide a new idea in studying pathogenesis of adipocyte differentiation.  [Key words]T3-L1 cell ; Gene expression ; cDNA microarray  脂肪组织不仅是被动的能量储存器官，而且是能够分泌多种激素类物质的内分泌器官；人类多种疾病与脂肪细胞分化及其调控失常有关，包括肥胖症、脂肪肝、高脂血症、糖尿病及乳腺癌等疾病[1]。研究脂肪细胞分化机制及其调控，不仅对研究上述生理和疾病过程机制有重要理论意义，而且对其预防与 治疗 也具有实际意义。为此, 我们应用基因芯片技术研究3T3-L1细胞分化前后基因表达谱的改变, 全面了解3T3-L1细胞分化所伴发的基因差异表达, 研究3T3-L1细胞分化的产生机制并为寻找相应的干预手段提供理论基础。  1 材料与方法  材料T3-L1细胞由本实验室保存，DMEM培养基、胎牛血清(fetal bovine serum，FBS)、胰蛋白酶和TRIzol由美国Invitrogen公司提供，胰岛素、地塞米松、1-甲基-3-异丁基黄嘌呤(MIX)和油红O(oil red-O)染料由美国Sigma公司提供，One-Cycle cDNA Synthesis Kit、 I