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目的 探讨去甲二氢愈创木酸 (NDGA)诱导人恶性胶质瘤细胞系SHG 44细胞分化过程中基因组甲基化状态改变及其意义。方法 合成甲基化敏感性AP PCR引物 ,用AP PCR方法检测SHG 44细胞基因组甲基化状态 ,观察NDGA诱导该细胞分化过程中甲基化水平的变化。结果 对照组基因组DNA经MspⅠ酶解后AP PCR扩增片段比HpaⅡ酶解后的扩增片段小 ,几乎未见存留大片段 ,至少有 3个片段与HpaⅡ酶解片段不同。同一时相点HpaⅡ酶解后扩增产物相比 ,NDGA处理后基因组DNAMspⅠ酶解后的扩增产物量较少、条带较多 ,且随时相点延长产物量逐渐增加 ,条带增多。结论 NDGA诱导人恶性胶质瘤细胞系SHG 44细胞分化过程中基因组甲基化状态增高 ,推测基因组甲基化状态可能是NDGA诱导胶质瘤细胞分化的作用靶点之一
Objective To investigate the genomic methylation status of human glioblastoma cell line SHG 44 induced by nordihydroguaiaretic acid (NDGA) and its significance. Methods Methylation sensitive AP PCR primers were synthesized. The methylation status of SHG 44 cells was detected by AP PCR. The changes of methylation level during NDGA induced differentiation were observed. Results In the control group, the AP PCR amplified fragment was smaller than the HpaII digested fragment after MspⅠ digestion. There were almost no large retained fragments and at least 3 fragments were different from the HpaII digested fragment. Compared with the amplification product after HpaII digestion at the same time point, the amount of amplified product after enzymatic digestion of genomic DNA with NDGA treatment was less, the bands were more, and the amount of product was gradually increased and the bands increased at any time. Conclusion NDGA induces methylation of genomic DNA during the differentiation of human glioblastoma cell line SHG-44. It is speculated that methylation status may be one of the targets of NDGA-induced glioma cell differentiation