基因工程抗体增强紫杉醇诱导乳腺癌细胞SKBr3凋亡的作用  

Enhanced sensitization to paclitaxel-induced apoptotic effect on breast cancer cells SKBr3 by engineered antibody treatment

作　　者：尹荟菁[1] 程联胜[2] 

机构地区：[1]安徽农业大学生命科学学院分子遗传学实验室,合肥230036 [2]中国科学技术大学生命科学学院细胞与分子免疫学实验室,合肥230027

出　　处：《中国免疫学杂志》2007年第3期229-233,237,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(30400078);安徽省自然科学基金(050430201)资助

摘　　要：目的探讨抗p185c-erbB-2/neu基因工程抗体增强化疗药物紫杉醇诱导p185高表达的人恶性乳腺癌细胞系SKBr3凋亡的效应及其机制。方法采用MTS法检测基因工程抗体、紫杉醇及两者联合对SKBr3细胞增殖的抑制作用;用AnnexinV-FITC/PI法和流式细胞术定量分析SKBr3细胞的凋亡率;用Western blot技术分别检测AKt的Ser473位点、NF-κBp65亚基的磷酸化水平;用EMSA分析SKBr3细胞核内NF-κB与DNA的结合活性。结果基因工程抗体可增强紫杉醇对SKBr3细胞增殖的抑制作用,并可诱导细胞凋亡;基因工程抗体联合紫杉醇并可显著抑制SKBr3细胞中AKt/NF-κB途径。结论紫杉醇诱导乳腺癌细胞SKBr3凋亡的同时,可能通过激活AKt/NF-κB途径使癌细胞对紫杉醇产生耐药性。抗p185c-erbB-2/neu基因工程抗体联合紫杉醇是通过抑制AKt/NF-κB途径而增强紫杉醇诱导癌细胞凋亡的作用。Objective： To explore the enhanced sensitization to chemotherapeutic agent paclitaxel-induced apoptotic effect on p185-overexpressing human malignant breast cancer cell lines SKBr3 by anti-p185^c-erbB-2/neu engineered antibody treatment and to study its emerging mechanism. Methods：The proliferative inhibitory effect was assessed by MTS assay; Cells stained with AnnexinV-FITC and PI were used to qualify the apoptotic cell number by FACS （ Fluorescence-activated cell sorting） analysis; Phosphorylation of Ser473 AKt and p65 NF-κB subunit were determined by Western blot; NF-κB-DNA binding activity was demonstrated by EMSA（ Electrophoretic mobility shift assay）. Results：Anti-p185^c-erbB-2/neu engineered antibody rendered SKBr3 cells more susceptible to paelitaxel-induced proliferative inhibition, which further attributed to apoptotic induction; Furthermore, combination of the engineered antibody and paclitaxel also showed effective suppression of AKt/NF-κB pathway in SKBr3 cells. Conclusion：The aggressiveness of AKt/NF-κB pathway was partly attributed to its resistance to paclitaxel-induced apoptosis. Anti-p185^c-erbB-2/neu engineered antibody plus paclitaxel combination rendered p185-overexpressing human malignant breast cancer cells SKBr3 more susceptible to paclitaxel-induced apoptosis by efficient suppression of AKt/NF-κB pathway.

关 键 词：抗p185^c-erbB-2/neu基因工程抗体 紫杉醇 凋亡 AKt/NF-κB途径 恶性乳腺癌 

分 类 号：R392[医药卫生—免疫学]