5F11-DOX结合物对人肺腺癌细胞体内外抑制作用的研究  

Inhibition of human lung adenocarcinoma cells by 5F11-doxorubicin immunoconjugate in vitro and in vivo

作　　者：张同梅[1] 赖百塘[1] 湛秀萍[1] 

机构地区：[1]北京市结核病胸部肿瘤研究所

出　　处：《中国肺癌杂志》2005年第6期495-500,共6页Chinese Journal of Lung Cancer

基　　金：北京市科委高技术实验室资助(No.954830600)~~

摘　　要：背景与目的随着抗体技术的发展,越来越多的抗体复合物用于肿瘤的治疗。本研究试图探讨单克隆抗体化疗药物结合物5F11DOX在体内外对人肺腺癌A2细胞的抑制作用及其可能的作用机制。方法戊二醛法制备5F11DOX结合物,集落形成实验观察结合物对肿瘤细胞的体外杀伤作用,荧光显微镜观察DOX在肿瘤细胞中的分布,免疫组化检测抗体与肿瘤细胞的结合部位及分布。裸鼠皮下或腹腔内接种A2细胞异种移植瘤,比较5F11DOX与游离DOX的疗效。结果浓度为0.04mg/L的5F11DOX即可在体外杀死全部人肺腺癌A2细胞,5F11DOX是游离DOX作用的10倍。荧光镜检发现在DOX浓度为3mg/L时作用3h,去除药物,继续培养24h后,5F11DOX组的DOX荧光较游离DOX组强。免疫组化检测发现5F11定位于细胞膜与胞浆中,而荧光镜检发现DOX分布于细胞内。裸鼠皮下及腹腔内异种移植瘤治疗实验显示5F11DOX组裸鼠的移植瘤明显小于对照组和同剂量游离DOX组(P<0.05);5F11DOX疗效相当于4～8倍的游离DOX。移植瘤的病理切片HE染色显示,5F11DOX结合物组癌巢中央和周围均有大片的瘤组织坏死。结论5F11DOX结合物对人肺腺癌A2细胞的杀伤作用显著高于游离的DOX。Background and objective With the development of antibody technology, more and more immunoconjugates are used in clinical treatment for different cancers. The aim of this study is to investigate the inhibitive effects of 5F11-DOX immunoconjugate on human lung adenocarcinoma cell line LTEP-A2 in vitro and in vivo and to explore the potential mechanism. Methods The 5F11-DOX immunoconjugate was produced by diluted glutaraldehyde crosslinking. The killing efficiency of 5F11-DOX was detected by clonogenic assay. The distribution of DOX. was ohserved under fluorescence microscope and the 5F11 location was determined by immunohistochemistry. The therapeutic efficacy of 5F11-DOX and free DOX was detected on subcutaneous or intraperitoneal exnogenic transplanted tumors of human lung adenocarcinoma A2 cells in nude mice. Results 55F11-DOX of 0.04 mg/L could kill all the A2 cells in vitro and the killing efficiency was 10 times as that of the free DOX. Fluorescence microscopy showed that fluorescence of DOX in 3 mg/L 5F11-DOX group was much stronger than that in 3 mg/l. free DOX group after treating A2 cells with 3 mg/L 5F11-DOX or DOX for 3 h, then incubating the cells with fresh medium for another 24 hours. Immunohistochemistry showed that 5F11 located in cell membrane and cytoplasm and fluorescence microscopy proved that DOX located inside the cells. The average sizes of subcutaneous or intraperitoneal exnogenic transplanted tumors in 5F11-DOX group were obviously smaller than those of the control group and free DOX group at the same dosage （P〈0.05）, and the anti-tumorogenicity efficacy of 5F11-DOX was 4-8 times as that of free DOX. The HE staining showed that extensive necrosis occurred in the center of tumors and around cancer nests in 5F11-DOX group. Conclusion The killing efficacy of 5F11-DOX on human lung adenocarcinoma cell line A2 is obviously higher than that of the free DOX.

关 键 词：肺肿瘤 单克隆抗体 阿霉素 免疫结合物 

分 类 号：R734.2[医药卫生—肿瘤]