【作者】 凌晓光；

【导师】 吴强； 程联胜； 孟刚；

【作者基本信息】 安徽医科大学 ， 病理学与病理生理学， 2006， 硕士

【摘要】 目的：观察抗HER-2工程抗体chA21对高表达HER-2的人卵巢癌SKOV3裸小鼠移植瘤的抑制作用，并初步探讨其可能的作用机制。方法：成功建立人卵巢癌SKOV3裸小鼠移植瘤模型后，随机将荷瘤裸鼠分为阴性对照NS组12只，chA21组6只和阳性对照药Herceptin组6只共3组。chA21和Herceptin均按30mg·kg^-1体重每周两次静脉注射，观察肿瘤生长变化，连续给药6周后，处死全部荷瘤裸鼠，计算抑瘤率；观察肿瘤组织的病理变化，AB／PAS染色观察肿瘤组织中黏液的分泌；应用HE染色、透射电镜和TUNEL等技术观察chA21对肿瘤细胞凋亡的影响；并利用组织芯片及免疫组化方法结合显微图像分析系统定量检测肿瘤组织中：1)细胞增殖指标Ki-67和NFκB的表达；2)凋亡相关基因Bax、Bcl-2、caspase 3及Fas蛋白的表达；3)与侵袭相关因子MMPs、TIMPs、VEGF及E-cad的表达。结果：chA21以30mg·kg^-1体重每周2次尾静脉注射给药：1)能显著抑制高表达HER-2的人卵巢癌SKOV3细胞裸小鼠移植瘤的生长，其抑瘤率为30.53％。2)能下调移植瘤细胞Ki-67和NFκB的表达。3)HE染色光镜观察chA21组移植瘤细胞出现聚巢倾向，并有腺样结构形成趋势，AB／PAS染色见细胞内AB阳性物质增多；细胞膜表面的E-cad表达增强。4)HE染色和透射电镜均能观察到chA21组移植瘤细胞出现凋亡现象；且chA21组移植瘤细胞凋亡指数显著高于NS组。5)chA21组移植瘤细胞Bax、Fas和caspase 3的表达增加，Bcl-2和NFκB的表达及Bcl-2／Bax比值下降。6)chA21组移植瘤细胞MMP-2、MMP-9和VEGF的表达下降，而TIMP-1、TIMP-2及E-cad的表达增加，MMP-2／TIMP-2和MMP-9／TIMP-1的比值下降。结论：1)抗HER-2工程抗体chA21能够抑制人卵巢癌SKOV3裸小鼠移植瘤的生长。其可能的机制是通过抑制NFκB的表达，下更多还原

【Abstract】 Objective To study the anti-tumor effect of anti-HER-2 engineered antibodies chA21 on nude mice xenografts of human ovarian cancer SKOV3 cells and explore its mechanism. Methods An animal model with human ovarian cancer SKOV3 cells involved in nude mice was established and the mice were randomized into 3 groups: 12 in normal saline group （negative control）, 6 in chA21 group and 6 in Herceptin （positive control）. The mice were administrated respectively with chA21 (30mg·kg^-1) and Herceptin (30mg·kg^-1) via caudal vein injection twice a week for consecutive 6 weeks, and then were killed after 44 day’s administration of the drugs. The volumes of the xenografts were measured twice a week. The tumor weight and inhibition ratio were measured after mice were killed. Pathological changes of tumor tissues were observed. AB/PAS staining was used to evaluate the mucus production. HE staining, electron microscopy and TIJNEL staining were used to detect the apoptosis induction by chA21 in SKOV3 cells. Expression of Ki-67, NFκB, Bax, Bcl-2, caspase 3, Fas, MMP-2, MMP-9, MMP1, TIMP-2, VEGF and E-cad in the three groups was quantificationally analyzed by immunohistochemistry （S-P method） on tissue microarray sections combined with a micro-image analyse system. Results 1) The growth of xenografts of human ovarian cancer SKOV3 cells in nude mice was significantly inhibited by chA21 with 30.53% of the tumor inhibitory ratio. 2) Both Ki-67 labeling index and NFkB protein expression in chA21 group were lower than those in the negative control. 3) A tendency of tumor cell nests and glandular formation in chA21 group was observed by更多还原