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目的观察高表达热休克蛋白70(HSP70)C6细胞瘤苗激活的SD大鼠脾细胞杀伤靶细胞的超微结构变化,并初步探讨其可能的抗瘤机制。方法采用经诱导高表达HSP70的灭活C6细胞作瘤苗,体外刺激大鼠脾细胞进行肿瘤杀伤试验。采用噻唑蓝(MTT)法检测其杀伤活性,流式细胞术(FCM)及电子显微镜观察杀伤瘤细胞的变化。结果(1)经瘤苗刺激的大鼠脾细胞对C6细胞的杀伤率较直接用灭活C6细胞刺激的脾细胞或未受任何刺激的新鲜脾细胞显著增高。(2)行肿瘤杀伤试验时FCM检测到亚二倍体峰,电镜发现靶细胞受攻击后出现染色质浓聚于核膜边缘,呈境界分明的块状或月形、半月形小体,或整个细胞核固缩成块状物,电子密度高,核膜、胞膜完整,或可见到凋亡小体,部分见髓鞘形成。有些效应细胞坏死。结论介导靶细胞凋亡可能是高表达HSP70的C6细胞瘤苗主动免疫抗瘤效应的一个重要机制。
OBJECTIVE: To observe the ultrastructural changes of splenocyte-killing target cells in SD rat spleen cells activated by heat shock protein 70 (HSP70) -cells and to explore its possible anti-tumor mechanism. Methods Inactivated C6 cells induced by high expression of HSP70 were used as tumor vaccine, and spleen cells were stimulated in vitro for tumor killing test. The cytotoxic activity was detected by MTT assay. The changes of cytotoxicity were observed by flow cytometry (FCM) and electron microscopy. Results (1) The killing rate of C6 cells by the spleen-stimulated rat spleen cells was significantly higher than that of the spleen cells stimulated by the inactivated C6 cells or the spleen cells without any stimulation. (2) Sub-diploid peaks were detected by FCM in tumor killing assay. Electron microscopy showed that chromatin was clustered on the edge of nuclear membrane after being attacked by target cells, which was characterized by massive or crescent moon, meniscus, or The entire nucleus shrinks into lumps, high electron density, nuclear membrane, cell membrane integrity, or apoptotic bodies can be seen, in part, see myelination. Some effector cells are necrotic. Conclusion Apoptosis mediated by target cells may be an important mechanism of active immune antitumor activity of C6 cell vaccine expressing HSP70.