摘要
目的:观察去甲二氢愈创木酸(nordihydroguaiareticacid,NDGA)对人恶性胶质瘤细胞系SHG-44细胞凋亡的影响,并探讨其发生的可能机理。方法:采用MTT(methythiazolyltetrazolium)法检测细胞增殖抑制的情况,利用光镜和电镜观察及TUNEL法检测体外培养细胞的凋亡情况,用免疫组织化学、原位杂交和图像分析等方法检测bcl-2基因mRNA和蛋白的表达水平。结果:①一定浓度(100μmol/L)的NDGA处理SHG-44细胞不同时间后,在NDGA抑制细胞增殖的同时,也可诱导此细胞发生凋亡,且随着作用时间的延长,凋亡细胞数增加越明显;②免疫组化染色结果表明,一定浓度的NDGA处理细胞不同时间后,出现SHG-44细胞bcl-2蛋白表达水平降低,且随着作用时间的延长,这种趋势更加明显,与细胞凋亡的发生呈负相关;③原位杂交结果显示,NDGA处理细胞不同时间后,出现SHG-44细胞bcl-2基因mRNA的表达水平降低,结果与免疫组化检测一致。结论:NDGA可诱导SHG-44细胞的凋亡,这种作用可能与其下调bcl-2基因的表达有关,确切机制有待进一步深入研究。
Objective: To investigate the effects and probable mechanisms of nordihydroguaiaretic acid (NDGA) on apoptosis in human malignant glioma cell line SHG-44. Methods: Cell growth inhibition was measured with MTT method. Cell apoptosis was observed with light and electron microscopy and TUNEL. Expression of bcl-2 gene was measured with immunohistochemistry , in situ hybridization, and image analysis. Results: ①NDGA at the concentration of 100 μmol/L inhibited the proliferation of SHG-44 cells and induced apoptosis, and this effect was positively correlated to the duration of NDGA treatment. ②The immunohistochemistry results indicated that protein expression of bcl-2 gene in SHG-44 cells decreased after treatment with 100 μmol/L NDGA and it was negativelg correlated to the duration of NDGA treatment and the degree of cell apoptosis. ③In situ hybridization results indicated that mRNA expression of bcl-2 gene in SHG-44 cells decreased after treatment with 100 μmol/L NDGA and it was apparently consistent with the immunohistochemistry results. Conclusion: NDGA can induce apoptosis of human malignant glioma cells and this effect is probably performed by down regulation expression of bcl-2 gene, the exact mechanism needs further study.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2001年第11期1246-1250,共5页
Chinese Journal of Cancer
基金
国家自然科学基金资助项目(NO.39670296)
