摘要
目的探讨免疫效应细胞促进阿霉素(ADR)诱导乳腺癌耐药细胞MCF7/ADR凋亡的作用机制。方法采用干扰素γ(IFNγ)、CD3单抗、白介素(IL)1和IL2诱导并扩增免疫效应细胞。利用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)技术和P糖蛋白(P gp)免疫组织化学染色,观察P gp表达与凋亡的关系。用流式细胞术检测乳腺癌相关抗原P185蛋白的表达,以及AnnexinV FITC/PI阳性率。荧光显微镜下观察ADR在靶细胞内的分布和AnnexinV的表达。结果免疫效应细胞使MCF7/ADR细胞P185和P gp表达明显下降,CIK细胞作用后,MCF7/ADR细胞P185标记荧光几何均数下降了91.9%。免疫效应细胞增加了ADR在MCF7/ADR细胞内的积累及其在细胞核的分布。联合应用免疫效应细胞和ADR后,MCF7/ADR细胞凋亡率达78.9%,比单纯加入ADR组增高了10倍,较单纯加入免疫效应细胞组增高了13倍。结论免疫效应细胞可同时下调P185蛋白和P gp蛋白的表达,协同ADR可提高MCF7/ADR细胞的凋亡率。
Objective To investigate the effects of immunologic effector cells to enhance apoptosis induced by adriamycin ( ADR ) in multi - drug resistant human breast cancer cell line MCF7 / ADR. Methods The immunologic effector cells were induced and expanded by IFN-gamma, McAb CD3, IL-1 and IL-2. The expression of P-glycoprotein (P-gp) and its relation to apoptosis in target cells were detected by TUNEL technique and immunohistoehemical staining. Flow cytometry (FCM) was carried out to determine the expression level of human breast cancer related P185 antigen and the positive rate of Annexin V-FITC/PI expression. The subcellular distribution of ADR and Annexin V expression in the target cells were detected by fluorescence microscopy. Results The immunologic effector cells down-regulated the expression of P185 and P-gp in MCF7/ADR cells. The accumulation and subcellular distribution of ADR in MCF7/ADR cells were increased after co-culture with the immunologic effector cells. After treatment with the immunologic effector cells in combination with ADR, apoptosis rate of the target cells was 10 times higher than that induced by ADR alone, and 13 times higher than that induced by the immunologic effector cells alone. Conclusion Immunologic effector cells can simultaneously down-regulate the expression of P185 and P-gp in MCF7/ADR cell line, and increase the apoptosis rate of MCF7/ADR cells in combination with ADR.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2006年第3期188-191,共4页
Chinese Journal of Oncology
关键词
免疫细胞
多药耐药
P-糖蛋白
阿霉素
乳腺肿瘤
Immunocyte
Multidrug resistance
P-giycoprotein
Adriamycin
Breast neoplasms