摘要
目的：研究多种细胞因子诱导的杀伤细胞（cytokine-induced killing cells,CIK）对乳腺癌细胞株ZK-75-1的杀伤作用,并探讨其作用机制.方法：通过HE染色观察凋亡细胞ZK-75-1的形态学改变.应用TUNEL（TdT-mediated dUTPnick end labeling）法检测CIK细胞的凋亡.通过免疫细胞化学染色法检测ZK-75-1细胞中p53、p16、C-myc、Bcl-2及Bax的表达率.结果：HE染色显示,CIK细胞向ZK-75-1细胞靠近,形成典型的玫瑰花环状;肿瘤细胞的胞浆中出现颗粒状物,有的肿瘤细胞只见颗粒状碎片;而作为对照的乳腺癌细胞生长良好.TUNEL法检测显示,对照组细胞未染色或染呈均匀的淡蓝色;实验组凋亡的细胞缩小,核或核周染呈深蓝色.CIK细胞作用4～12 h ZK-75-1细胞的凋亡率上升,作用12～24h细胞的凋亡率下降,与对照组相比较有统计学意义（P＜0.01）.免疫细胞化学染色的结果表明,CIK细胞实验组p53、pi6、C-myc及Bcl-2蛋白随作用时间的延长均下降,Bax蛋白的表达上调,与对照组相比较均有统计学意义（P＜0.01）.结论：CIK细胞对乳腺癌细胞ZK-75-1杀伤作用的机制之一,可能与p53、p16、C-myc、Bcl-2蛋白表达的下调及Bax蛋白表达的上调有关,并与CIK细胞作用的时间关系密切.
AIM： To study the cytotoxicity and mechanism of cytokine-induced killer （CIK） cells to breast cancer cell line ZK-75-1. METHODS： The morphological changes of ZK-75-1 cells were observed by HE staining. The apoptosis of ZK-75-1 cell line was examined by TUNEL staining. The expression of P53, P16, C-myc, Bcl-2 and Bax were determined by immunocytochemical staining. RESULTS： The result of HE staining revealed that CIK cells moved toward the target cells, forming typical rosette cells. Granulelike substances appeared in cytoplasm of tumor cells, but only granule-shape patch was found in some tumor cells while, breast cancer cells as control grew well. TUNEL staining indicated that the cells in control group were not stained or dyed well-distributed light blue. As the cells in experiment group became smaller, mucleoluser or perinuclear were dyed deep blue. The apoptotic rate of ZK-75-1 cells cocultured with CIK cells was increased after 4 -12 hours and was decrensed after 12 - 24 hours, with a significant difference compared with control group （ P〈0.01 ）. Immunocytochemical staining showed that the expression of 1353, p16, C-myc and Bcl-2 proteins in CIK group declined but the expression of Bax protein increased with the passage of time, which was significantly different compared with control （ P 〈 0.01 ）. CONCLUSION： The mechanism of killing ZK-75-1 cells by CIK cells is closely related to the downregulation of the expression of P53, P16, c-myc and Bcl-2 proteins and to the upregulation of the expression of Bax protein. It also has close relotion with the time of exposure to CIK cells.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2006年第3期314-317,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
吉林省科学技术厅资助课题(No.20010557)
