三种反义c-myc RNA稳定表达对血管平滑肌细胞生长、增殖的影响  

Effects of three kinds of stable expressive antisense c-myc RNA on the growth and proliferation of vascular smooth muscle cell

作　　者：曾嵘[1] 刘小蓉[1] 李进[1] 安靓[1] 戴云[1] 

机构地区：[1]第一军医大学组胚教研室,广东广州510515

出　　处：《第一军医大学学报》2000年第6期499-502,共4页Journal of First Military Medical University

基　　金：国家自然科学基金资助项目!(39900179)

摘　　要：目的 研究反义基因在细胞中持续、稳定表达对平滑肌细胞生长增殖的影响。方法 将分别载有c(myc第1、2和3外显子反义片段的三种重组逆转录病毒表达载体aM1、aM2和aM3导入大鼠主动脉平滑肌细胞（SMC），并于持续筛选后1个月进行稳定表达检测。结果 aM1和aM2能持续抑制靶细胞Myc蛋白表达（分别为对照表达量的59.7%和81.3%）、增殖细胞核抗原（PCNA）表达（分别为对照表达量的52.4%和51.9%），抑制靶细胞的生长增殖活性并以aM2作用最强（抑制率达到30%），同时，aM2还有一定的延长细胞周期和减少DNA合成的作用，而aM3作用相反。另外，与瞬时表达相比，外源基因在靶细胞中的作用并非一成不变，而基因转染操作对细胞的影响会随着细胞的代偿逐渐减少并在短期内消失。结论 对c(myc中不同区域进行反义封闭会产生不同的效应，对其转录活性区进行封闭可取得最强的细胞生长抑制。而靶细胞对对外源基因导入可能具有“纠正作用”。Ojective In order to understand the effects of continuous and stable expression of antisense genes on the growth and proliferation of the smooth muscle cells. Method Three combinant retrovial expression vectors, aM1, aM2 and aM3, which was respectively loaded with reverse fragment of c-myc exon 1, 2 and 3, were transferred into rat arterious smooth muscle cells and assayed 1 month after their stable expression. Result aM1 and aM2 could inhibit Myc expression (respectively reduced to 59.7%, 81.3%), and the expression of proliferat cellular nuclear antigen (PCNA) (respectively reduced to 52.4%, 51.9%). aM1 and aM2, especially the latter also inhibited cellular proliferative activity, aM2 slightly hindered cellular cycle and DNA synthesis too, while the effects of aM3 turned out to be contrary to that of aM1 and aM2. In addition, compared with transient expression, the effects of introduced genes were variable in target cells, and the changes brought about by the transfection process would reduce and disappear in a short time with the lowering of cellular compensation. Conclusions Blocking different regions of c-myc can achieve different effects and the blokade of the transcription activity regions usually produces enhanced growth inhibition effects. Moreover, target cells may have some 'correction mechanism ' against the introduced foreign genes.

关 键 词：血管平滑肌细胞 基因治疗 C-MYC RNA SMC 

分 类 号：R392[医药卫生—免疫学]