【摘要】 研究人外周血树突状细胞(dendritic cell,DC)联合LPAK(lymphokine and PHA activatedkiller)细胞体外诱导人肝癌细胞株BEL-7402凋亡的作用,并进行形态学分析。实验分LD(BEL-7402+LPAK+DC),L(BEL-7402+LPAK),D(BEL-7402+DC)和 B(BEL-7402)四组,采用中性红摄入比色法检测细胞毒活性,原位末端标记(TUNEL)技术、电镜技术观察细胞的光镜形态和超微结构。结果显示,D组与B组比较吸光度值无显著性差异(P>0.05),即无杀伤活性;LD组与L组比较,细胞毒活性为LD组>L组(P<0.01)。光镜下凋亡的肿瘤细胞呈TUNEL阳性,细胞核染成棕黄色或深棕色,大小不等,形态也不一;电镜下肿瘤细胞核染色质凝聚、固缩和胞质浓缩,凋亡小体多见;少部分LPAK细胞内形成自噬体,发生自噬性凋亡。结论提示,人血DC联合LPAK细胞能有效地诱导肝癌细胞凋亡,在此过程中LPAK细胞同时发生自噬性凋亡。更多还原

【Abstract】 To observe the effects and morphological changes of human peripheral blood dendritic cells (DC) on lym-phokine and PHA activated killer (LPAK) cells inducing apoptosis of human hepatoma cell line (BEL-7402) in vitro, the experimental groups were divided into LD group (BEL-7402 + LPAK + DC), L group (BEL-7402 + LPAK), D group ( BEL-7402 + DC) and B (BEL-7402) group. The methods of neutral red uptake, light mi- croscopy, electron microscopy and TDT mediated X-dUTP nick ending labeling (TUNEL) were adapted in out experiment. The results showed as follows: ① the difference between D group and B group was not distinct (P >0.05), and the difference between LD group and L group was distinct, DC+ LPAK>LPAK (P<0.01) in cytotoxity; ② The apoptotic cells were TUNEL positive in light microscopy, and the apoptotic nuclei were stained yellow brown and dark brown, which varied in size and shape from cell to cell; ③ The ultrastuctural change of apoptotic tumor cells comprised compaction and condensation of nuclear chromatin, condensation of cytoplasm and apoptotic bodies; at the same time, LPAK cells manifested the characteristics of autophagic apop-tosis, and there were some autophagic bodies in it. It is concluded that the combination of human blood DC and LPAK cells could induce apoptosis of BEL-7402 cells effectively, in which some LPAK cells manifested the characteristics of autophagic apoptosis. Department of Histology & Embryology, Shantou University Medical College Shantou 515031 Department of Anatomy Hongkong Chinese University, Hongkong更多还原