冷冻诱导大鼠C6脑胶质瘤细胞发生凋亡的实验研究

目的:通过观察大鼠C6脑胶质瘤经冷冻后出现细胞凋亡,探讨冷冻对肿瘤的杀伤机制。方法:建立大鼠C6脑胶质瘤颅内接种模型,通过TUNEL法及流式细胞仪分别检测其冷冻后凋亡数目及凋亡率。结果:肿瘤经冷冻后,其中心出现坏死,而周边细胞呈现典型的凋亡形态学改变。TUNEL法检测冻后24、48和72h凋亡数目0.323、0.00167和0.037;流式细胞仪检测其冻后24、48和72h凋亡率与对照组相比,P值分别为0.0254、0.0064和0.031;冻后各时间点凋亡数目及凋亡率均显著高于对照组,P<0.05。其中冻后48h改变尤其明显,P<0.01,峰值时间出现在冷冻24～48h。结论:诱导细胞凋亡也是冷冻杀伤肿瘤细胞的重要机制之一,但其发生的分子机制有待于进一步深入研究。

Experimental study of apoptosis in C6 rat glioma model induced by cryotherapy

OBJECTIVE:To explore the mechanism of killing tumor cells by observing the C6 rat glioma cell apoptosis after cryotherapy. METHODS: By establishing the C6 rat glioma models, after cryotherapy, TUNEL and Flow cytometry were employed to inspect the number of apoptotic cells and apoptotic rate respectively. RESULTS: Cryotherapy can induce cell apoptosis. Necrosis was shown in the central of tumor cells and apoptosis was observed in the peripheral zone. The number of apoptotic cells detected by TUNEL method at 24, 48 and 72 h after freezing, which compared with that in the control group, P value was 0.323, 0.001 67 and 0.037 respectively. The apoptotic index detected by Flow cytometry at 24, 48, 72 h ours after freezing, which compared with that in the control group, P value was 0.025 4, 0.006 4 and 0.031 respectively, All the difference were statistical significance, P<0.05. This change was particularly evident at 48 h after freezing, P<0.01, The apoptotic peak was occurred during 24-48 hours after cryotherapy. CONCLUSIONS: Inducing apoptosis was also one of the mechanisms of killing tumor cells treated by cryotherapy. But the molecular mechanism of Induced apoptosis must be further investigated.