重组腺病毒介导的人p27kip1基因高表达对体外培养的大鼠主动脉血管平滑肌细胞迁移的影响

目的　探讨重组腺病毒介导的p2 7kip1基因及其蛋白产物高表达对血管平滑肌细胞 (VSMCs)迁移的抑制作用。方法　将含人p2 7kip1cDNA的重组腺病毒 (Adhp2 7kip1)及含 β 半乳糖苷酶基因的重组腺病毒 (AdLacZ)在体外转染原代大鼠主动脉VSMCs,用Westernblot及Boyden趋化小室检测外源性p2 7kip1蛋白在细胞内的表达及对VSM Cs迁移的影响。结果　转染后 2 4h ,Adhp2 7kip1转染的VSMCs内p2 7kip1蛋白高表达 ,而AdLacZ转染的VSMCs内仅显示极低水平的内源性p2 7kip1蛋白表达 ;Boyden趋化小室检测显示未转染的VSMCs、AdLacZ及Adhp2 7kip1转染的VSMCs血清诱导后的迁移细胞数分别为 139± 2 6、10 6± 16及 6 8± 14。结论　外源性p2 7kip1基因及蛋白产物在VSMCs内高表达可显著抑制VSMCs的迁移

Effects of recombinant adenovirus-mediated hmnan p27kip1 gene overexpression on migration of cultured primary rat aortic vascular smooth muscle cells

Objective To study the effect of adenovirus-mediated transduction of human p27kip1 gene and its protein product over-expression on migration of rat primary cultured aortic vascular smooth muscle cells(VSMCs).Methods Human p27kip1 recombinant adenovirus(Adhp27kip1) or LacZ recombinant adenovirus(AdLacZ) was used to infect rat primary cultured aortic vascular smooth muscle cells.The exogenous p27kip1 gene expression in transfected VSMCSs was examined by Western blot assay and its effect on migration was tested with Boyden's chamber method using microchemotaxis chambers.Results Twenty-four hours after infecting VSMCSs, p27kip1 protein was over-expressed in VSMCSs infected by Adhp27kip1,and the level of endogenous p27kip1 protein expression was much lower in VSMCs infected by AdLacZ or uninfected.Boyden's chamber method showed that, after serum stimulation, the number of migration of uninfected VSMCs,and VSMCs infected by AdLacZ and Adhp27kip1 were 139±26,106±16 and 68±14, respectively.Conclusion Adenovirus-mediated p27kip1 gene and its protein product over-expression can inhibit serun-stimulated migration of VSMCs significantly.