外源E2F和CArG顺式元件对血管平滑肌细胞表型相关基因表达的影响  

Effect of E2F and CArG Decoy Oligodeoxynucleotides on Expression of Phenotype-associated Genes of Vascular Smooth Muscle Cells

作　　者：韩梅[1] 温进坤[1] 

机构地区：[1]河北医科大学基础医学研究所生物化学研究室,石家庄050017

出　　处：《中国生物化学与分子生物学报》2001年第5期626-630,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助项目 (No .3 9970 2 74);河北省自然科学基金资助项目 (No .3 0 13 5 8)

摘　　要：利用转录因子“诱骗”策略 ,阻断血管平滑肌细胞 (VSMC)表型特异基因和增殖相关基因的反式激活 ,揭示VSMC表型转化和增殖之间的关系 .电泳迁移率改变分析结果表明 ,相当于分化型VSMC特异表达基因共有顺式元件CArG和细胞增殖相关基因共有顺式元件E2F的双股寡核苷酸(ODNs)可分别与从分化型和去分化型VSMC中提取的核蛋白特异性结合 ,形成DNA 蛋白质复合物 .Northern杂交结果显示 ,导入VSMC中的CArGODN可使平滑肌α肌动蛋白 (α actin)表达活性降低 ,肌丝数量减少 ,明显抑制转染细胞的再分化过程 .去分化型VSMC被E2FODN转染后 ,增殖相关基因c myc表达受到抑制 ,细胞增殖速率减慢 ,去分化表型特征减弱 .结果提示 ,E2F和CArG调控元件分别对VSMC增殖和分化起重要调节作用 ,并证实VSMC表型转化与增殖是两个密切相关但不完全相同的细胞事件 .The relationships between the phenotypic modulation and proliferation of vascular smooth muscle cells(VSMC) were studied by transcription factor“decoy”strategy in which the activation of VSMC\|specific expression genes and proliferation\|associated genes was blocked.The results of gel mobility shift assay showed that the nuclear extracts from differentiated VSMC could bind to CArG double\|stranded oligodeoxynucleotide(ODN),which is a consensus cis \|element for differentiated VSMC\|specific expression genes,and that those from dedifferentiated VSMC could interact with E2F ODN,which is a consensus cis sequence for the genes involved in cell cycle regulation.Transfection of redifferentiating VSMC with GArG ODN could inhibit the expression of smooth muscle α\|actin gene,decrease myofilament content,and prevent redifferentiating of VSMC.Transfection of dedifferentiated VSMC by E2F ODN might result in down\|regulation of \%c\|myc\% gene expression,inhibition of VSMC proliferation,and attenuation of phenotypic characteristics of dedifferentiated VSMC.These results suggest that E2F and CArG cis\|elements might play an important role in VSMC proliferation and differentiation.

关 键 词：顺式元件诱骗 表型转化 基因调控 血管平滑肌细胞 VSMC 表型相关基因 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R331.32[医药卫生—基础医学]