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目的:比较3种简易形态观察法检测肝癌细胞凋亡的效果。方法:用6%的乙醇作用人肝细胞癌HCC-9204细胞6 h,诱导其凋亡,进行未固定细胞的吖啶橙/溴化乙啶(AO/EB)双染色、细胞固定后的H-E染色,以及TUNEL法检测肝癌细胞凋亡的形态学特征。结果:用6%乙醇诱导肝癌细胞凋亡后,AO/EB、H-E染色法以及TUNEL法均能够不同程度地显示细胞变圆、细胞核深染和染色质边集、呈环状、团块或新月体状,以及细胞质浓缩和“出泡”现象等细胞凋亡的典型形态学特征。AO/EB双染色法能够显示呈致密浓染的黄绿色染色或出现黄绿色碎片的早期凋亡细胞,还能显示呈致密红染或出现红色碎片的晚期凋亡细胞,也能显示呈红色膨大状的坏死细胞。结论:AO/EB双染色、细胞固定后的H-E染色,以及TUNEL法等3种方法均可不同程度地观察到凋亡细胞的形态学变化,其中的AO/EB双染色法还可以区分早期凋亡、晚期凋亡和坏死细胞,具有较大的实用价值。
Objective: To compare the effects of three simple morphological observation methods on the apoptosis of hepatocellular carcinoma cells. Methods: Human hepatocellular carcinoma cell line HCC-9204 was induced by 6% ethanol for 6 h and then induced to apoptosis. Apoptotic cells were stained with acridine orange / ethidium bromide (AO / EB) Staining, and TUNEL assay of hepatocellular carcinoma apoptosis characteristics. Results: AO / EB, HE staining and TUNEL assay showed that the cells were round, nuclear stained and chromatin margined after being induced by 6% Month-old body shape, and the typical morphological features of apoptosis such as cytoplasmic condensation and “blistering” phenomenon. AO / EB double staining can show densely stained yellow-green staining or yellow-green fragments of early apoptotic cells, but also showed a dense red dye or red fragments of advanced apoptotic cells, can also be reddish enlarged Necrotic cells. CONCLUSION: AO / EB double staining, HE staining after cell fixation and TUNEL method can observe the morphological changes of apoptotic cells to varying degrees. The AO / EB double staining method can also distinguish the early apoptosis Dead, late apoptosis and necrosis of cells, with greater practical value.