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人血树突状细胞和LPAK细胞诱导肝癌细胞系凋亡的形态学体外研究

In vitro study on the morphology of human blood dendritic cells and LPAK cells inducing apoptosis of the hepatoma cell line.

作者信息

Sun J, Zhang J, Chen J, Chen H, Chew Y, Chen J

机构信息

Department of Allergy, Chinese Academy of Medcial Sciences, Peking Union Medical College Hospital, Beijing 100730, China.

出版信息

Chin Med J (Engl). 2001 Jun;114(6):600-5.

Abstract

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).

METHODS

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.

RESULTS

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 cytotoxicity. 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.

CONCLUSIONS

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.

摘要

目的

观察人外周血树突状细胞(DCs)对淋巴因子和植物血凝素(PHA)激活的杀伤细胞(LPAK)诱导人肝癌细胞系(BEL-7402,B)凋亡能力的体外影响及形态学变化。

方法

将实验组分为LD组(DCs + L + B)、L组(L + B)、D组(DCs + B)和B组。采用中性红摄取法、普通光学显微镜、电子显微镜、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)。

结果

D组与B组差异不明显(P > 0.05)。LD组与L组差异明显,细胞毒性方面DCs + LPAK > LPAK(P < 0.01)。光镜下凋亡细胞TUNEL阳性,凋亡细胞核染成黄棕色和深棕色,大小和形状因细胞而异。凋亡肿瘤细胞的超微结构变化包括核染色质浓缩和凝聚,细胞质凝聚及凋亡小体形成。同时,LPAK细胞表现出自噬性凋亡的特征,其中有一些自噬小体。

结论

人血液DCs与LPAK细胞联合能有效诱导BEL-7402细胞凋亡,部分LPAK细胞表现出自噬性凋亡的特征。

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