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目的:研究全反式维甲酸对C6胶质瘤细胞增殖和分化的影响。方法:用5 mg/L全反式维甲酸(all-trans-retinoic acid,ATRA)诱导C6胶质瘤细胞,通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)实验绘制细胞生长曲线以考察ATRA对细胞增殖的抑制作用;通过流式细胞仪、光镜、透射电镜和免疫组织化学染色鉴定细胞的分化状况。结果:C6胶质瘤细胞经全反式维甲酸诱导后,细胞增殖受到明显抑制,并且密度明显减少(P<0.01)。细胞周期受阻,G0/G1期延长,S期细胞比例明显减少(P<0.01)。光镜观察到诱导前的C6胶质瘤细胞呈正常的成纤维细胞形态,而诱导后的C6胶质瘤细胞变细长,中间呈圆形或卵圆形,两端形成长尖突起。流式细胞仪的凋亡率检测显示,未见明显的凋亡;透射电镜观察显示,细胞有早期凋亡改变,且胞浆内空泡增多,线粒体和内质网丰富,细胞结构基本趋向正常。C6胶质纤维酸性蛋白表达明显增强。结论:全反式维甲酸能抑制胶质瘤细胞的增殖,并能诱导C6细胞分化。
Objective: To investigate the effect of all-trans retinoic acid on the proliferation and differentiation of C6 glioma cells. METHODS: C6 glioma cells were induced by 5 mg / L all-trans-retinoic acid (ATRA) and treated with 3- (4,5-dimethylthiazol-2) The cell growth curve was drawn by phenyltetrazolium bromide (MTT) assay to investigate the inhibitory effect of ATRA on cell proliferation. Cell differentiation was identified by flow cytometry, light microscopy, transmission electron microscopy and immunohistochemical staining. Results: C6 glioma cells induced by all-trans retinoic acid, cell proliferation was significantly inhibited, and the density was significantly reduced (P <0.01). The cell cycle was blocked, the G0 / G1 phase was prolonged, and the proportion of S phase cells was significantly decreased (P <0.01). The C6 glioma cells showed normal fibroblast morphology before light irradiation, while the induced C6 glioma cells became slender and long, with round or oval in the middle and elongated cusps at both ends. Flow cytometry apoptosis rate showed no obvious apoptosis; transmission electron microscopy showed that the cells had early apoptosis, and cytoplasmic vacuoles increased mitochondria and endoplasmic reticulum, cell structure tends to normal . C6 glial fibrillary acidic protein expression was significantly enhanced. CONCLUSION: All-trans retinoic acid can inhibit glioma cell proliferation and induce C6 cell differentiation.