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机构地区:[1]吉林大学中日联谊医院神经外科,吉林长春130033 [2]广西北海市人民医院神经外科
出 处:《中国实验诊断学》2004年第1期12-15,共4页Chinese Journal of Laboratory Diagnosis
基 金:国家自然科学基金 (编号 3 0 2 713 3 4;3 990 0 15 2 )资助
摘 要:目的 观察诱导分化剂苯乙酸 (PA)抑制胶质瘤细胞C6增殖过程中 ,生物发育调节的主控基因———同源盒基因 (Hox基因 )表达的变化。方法 根据引物的不同 ,将已知的 2 2种HOX基因分为 3组 (P1,P2 ,P3)。用逆转录PCR(RT PCR)及图像分析法检测应用PA前后 ,Hox基因组在胶质瘤细胞C6中表达的变化。针对应用PA前后表达显著不同的Hox基因组 ,用组内包含的特异Hox基因的引物 ,重复上述实验 ,检测特异Hox基因的表达。Hox基因 (组 )表达水平用基因 (组 ) β 肌动蛋白 (β actin)灰度比值表示。结果 胶质瘤细胞C6中 ,P2组Hox基因表达明显弱于P1、P3组(0 6 82 5± 0 0 987<0 8817± 0 0 731,0 86 0 8± 0 0 881,P <0 0 0 1)。应用PA后 ,P2组Hox基因表达 (0 8386± 0 0 811)显著增强 ,与用药前比较 ,差异显著 (P <0 0 0 1)。P2组中 ,HoxB2基因应用PA后比未用药组表达显著增强 (0 7763± 0 12 4 1>0 4 839± 0 1343,P <0 0 0 1)。HoxB4基因在应用PA前后于胶质瘤细胞C6中均未见表达。结论 胶质瘤的发生可能与HoxB2基因表达减弱有关。PA在分化诱导胶质瘤过程中 ,对HoxB2基因mRNA水平的表达有明显的上调作用 ,PA的作用机理与调节胶质瘤细胞转录过程有关。Objective Research has demonstrated that phenylacetate(PA) can inhibit growth and induce differentiation in rat C6 glioma cell line. But the mechanisms of PA action are still incompletely understood. This study is aimed to identify which Hox gene is related to glioma and to observer the change of expression on mRNA level treated by phenylasetate. Methods 22 kinds of Hox gene divided into 3 groups according to their primer sequence. Semiquantitative reverse transcription-polymerase chain reaction(RT-PCR) used to investigate the mRNA expression of Hox gene groups and some kind of Hox gene in rat C6 glioma cell line following differentiation induced by PA. The level of Hox gene expression was expressed as ratio expression rate(RER) of Hox gene/β-actin according to Computer Image analysis and the difference between C6 cells and PA treated C6 cells was analyzed by student t-test. Results It was found that Hox genes matching to primers P2 were weakly expressed in C6 cells(0.682 5±0.098 7<0.881 7±0.073 1, 0.860 8±0.088 1, P<0.001), the expression of HoxB2 mRNA was significantly up-regulated in PA treated C6 cells(0.776 3±0.124 1>0.483 9±0.134 3, P<0.001). Conclusion It can be inferred from these findings that the weak expression of HoxB2 maybe involved in glioma origin and the mechanisms of PA action are correlated with transcription process in glioma cells.
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