树突状细胞联合LPAK细胞体外诱导肝癌细胞株凋亡的形态学研究

摘要

目的研究人外周血树突状细胞(Dendritic Cell，DC)联合LPAK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。rn方法实验分LD、L、D和B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。rn结果 D组与B组比较吸光度值无显著性差异，即无杀伤活性(P>0.05)；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。rn结论人血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。%Objective　To observe in vitro effects and morphological changes of human peripheral blood dendritic cells (DCs) on the ability of lymphokine and phytohaemagglutininum (PHA) activated killer (LPAK) cells to induce apoptosis of the human hepatoma cell line (BEL-7402， B).rnMethods　Experimental groups were divided into LD group （DCs+L+B）, L group （L+B）， D group （DCs+B） and B group. The methods of neutral red uptake, ordinary light microscopy, electron microscopy, TDT mediated X-dUTP nick end labeling (TUNEL) were used. rnResults　The difference between the D group and the B group was not distinct (P>0.05). The difference between the LD group and the L group was distinct， with DCs+LPAK >LPAK (P<0.01) in cytotoxity. Apoptotic cells were TUNEL positive in light microscopy, and apoptotic nuclei were stained yellow brown and dark brown, with size and shape varying from cell to cell. Ultrastructural change in apoptotic tumor cells comprised of compaction and condensation of nuclear chromatin， and condensation of cytoplasm and apoptotic bodies. At the same time, LPAK cells manifested the characteristics of autophagic apoptosis, and there were some autophagic bodies in it. rnConclusions　The combination of human blood DCs and LPAK cells could induce apoptosis of BEL-7402 cells effectively, with some LPAK cells manifesting the characteristics of autophagic apoptosis.