p27kip1基因转移抑制大鼠胶质瘤生长的实验研究

目的]探讨重组腺病毒介导的人突变型p27kip1基因（Ad-p27kip1）表达对大鼠神经胶质瘤移植瘤生长的抑制作用。方法]建立大鼠胶质瘤移植瘤模型,随机分成3组：Ad-p27组、Ad-LacZ组、PBS组。Ad-p27组给予腺病毒介导的人p27 kip1基因（Ad-p27kip1）瘤内注射,另2组分别给予Ad-LacZ和PBS瘤内注射,观察对肿瘤生长的抑制情况,并用免疫荧光法检测P27和CyclinD1的表达;原位末端标记（TUNEL）法检测细胞凋亡情况。结果]给予Ad-p27kip1基因治疗的实验组大鼠肿瘤生长受到明显抑制;在颅内肿瘤模型组给予Ad-p27治疗的大鼠生存时间明显延长;免疫荧光染色显示移植瘤P27表达明显增强,CyclinD1的表达受到抑制。Ad-p27组细胞凋亡显著多于Ad-LacZ组和PBS组。结论]腺病毒介导的人p27基因（Ad-p27kip1）对大鼠胶质瘤的生长具有明显的抑制作用,其机制是增强p27表达、抑制CyclinD1的表达、诱导胶质瘤细胞凋亡。

EXPERIMENTAL STUDY OF THE DEPRESSIVE EFFECT OF P27KIP1 GENE ON GLIOMAS IN RATS

Objective]To explore the depressive effect of recombinant adenovirus mediated human mutant p27kip1 gene(Ad-p27kip1)on glioma in rats.Methods]The glioma model was established by implanting glioma cells in rats.The transplanted glioma rats were randomly divided into Ad-p27 kip1 group,Ad-LacZ group and PBS group.Rats in Ad-p27 kip1 group were given intratumoral injection of Ad-p27 kip1,that rats in Ad-LacZ group and PBS group were given intratumoral injection of Ad-LacZ and PBS respectively.Inhibitory action was evaluated by growth curves and inhibitory rates.The expression of p27 kip1 and CyclinD1 were detected by the immunofluorescence technique.TdT-mediated d-UTP nick end labeling(TUNEL)was used to detect the apoptosis situation of glioma cells.Results]The growth of glioma in Ad-p27kip1 group was significantly suppressed.The survival time of rats with intracranial tumor in Ad-p27kip1 group prolonged obviously than that in Ad-LacZ group and PBS group.Immunofluorescence assay showed that the protein product of p27kip1 increased and the expression of CyclinD1 in glioma was suppressed in Ad-p27kip1 group.More apoptotic glioma cells were detected in Ad-p27kip1 group than in either Ad-LacZ group or PBS group.Conclusion]Ad-p27kip1 has a significantly depressive effect on the growth of xenografts derived from glioma cells.The mechanisms include up-regulating expression of p27kip1,down-regulating expression of CyclinD1,and inducing apoptosis of glioma cells.