摘要
目的 制备高效、低毒的抗肝癌单链免疫毒素。 方法 将人突变型肿瘤坏死因子α(mTNF α)与人源化抗肝癌单链抗体hscFv25基因连接,构建pGEx4T-1/hscFv25-mTNF α融合蛋白原核表达载体,在大肠杆菌中表达并纯化目的蛋白,经western blot鉴定之后,进行荷肝癌(SMMC-7721)裸鼠体内初步抑瘤实验,并对治疗后裸鼠肿瘤组织进行抗TNF α的免疫组织化学染色。 结果 原核表达载体pGEX4T-1/hscFv25 mTNF α融合蛋白的表达量占细菌总蛋白的12％,hscFv25-mTNFα治疗组对荷肝癌裸鼠的有效率为5/5,其中2/5为完全缓解,3/5为部分缓解,疗效明显高于mTNFα对照组(F=8.70,P<0.05),治疗组裸鼠的肝、肺等组织中未见转移性病灶,经hscFv25-mTNFα治疗后的肿瘤组织,呈TNFα弥漫阳性反应,阳性颗粒主要位于瘤细胞的胞质中。 结论 hscFv25-mTNFα是高效、低毒的抗肝癌单链免疫毒素。
Objective To obtain high therapeutic effect and low toxicity single-chain immunotoxin against hepatocellular carcinoma (HCC). Methods Human mutant tumor necrosis factor-alpha (mTNFα ) was linked with the 3' end of humanized single-chain Fv against HCC (hscFv25) in pGEX4T-1 vector. The anti-HCC immunotoxin was expressed in Escherichia coli and identified by western blot. The primary tumor regression trial in nude mice bearing HCC was evaluated the targeting therapeutic value of hscFv25-mTNFα. The tumor tissues were stained by immunohis-tochemical with TNFa antibody. Results The expression of single-chain immunotoxin hscFv25-mTNFα was 12% of total bacteria proteins. The tumor regression trials of hscFv25-mTNFα showed 5/5 effective. It had 2/5 completely remission and 3/5 partly remission. The therapeutic result of hscFv25-mTNFα was better than that of mTNFα (F = 8.70, P < 0.05). The HCC tissue treated by hscFv25-mTNF α expressed TNFα positive reaction. The positive granule mainly existed in HCC cytoplasm. Conclusion The single-chain immunotoxin hscFv25-mTNFα has high therapeutic effect and low toxicity. It has potentialities for clinical application.
出处
《中华肝脏病杂志》
CAS
CSCD
2004年第3期148-150,共3页
Chinese Journal of Hepatology
关键词
肝癌
单链抗体
免疫毒素
肿瘤坏死因子
制备
Carcinoma, hepatocellular
Single-chain Fv
Immunotoxin
Tumor necrosis factor
