针对人肺腺癌的c-erbB-2/p185导向治疗

c-erbB-2/p185-directed therapy in human lung adenocarcinoma.

作者信息

Snider J M, Bushnell L J, Chen L C, Lanza L A

机构信息

Division of Cardiothoracic Surgery, University of Iowa Hospitals and Clinics, Iowa City, USA.

出版信息

Ann Thorac Surg. 1996 Nov;62(5):1454-9. doi: 10.1016/0003-4975(96)00630-3.

DOI:10.1016/0003-4975(96)00630-3

PMID:8893583

Abstract

BACKGROUND

These experiments were conducted to determine whether p185 can be therapeutically targeted in adenocarcinoma of the lung using an anti-p185-gelonin conjugate. c-erbB-2/p185 is overexpressed in up to one third of non-small cell lung cancers. CALU-3 is a human lung adenocarcinoma cell line that overexpresses p185. muMoAb-4D5 is a murine anti-p185 monoclonal immunoglobulin G1. Gelonin is a potent type 1 ribosomal inhibitory protein.

METHODS

4D5 and gelonin were covalently modified and linked. Purification was confirmed by SDS-polyacrylamide gel electrophoresis. Dose-dependent cytotoxicity was quantified using 3H-thymidine uptake by CALU-3 cells after incubation with 4D5-gelonin conjugate or with control substances (4D5, gelonin, unconjugated 4D5 + gelonin, or control antibody MOPC-21).

RESULTS

The 4D5-gelonin conjugate showed a 50% inhibitory concentration of 3.5 x 10(-10) mol/L, but 4D5 alone demonstrated no cytotoxic effect. Gelonin and the unconjugated 4D5-gelonin mixture had one tenth the cytotoxicity of the 4D5-gelonin conjugate (inhibitory concentration = 6.5 x 10(-9) mol/L and 8.5 x 10(-9) mol/L, respectively). The conjugate exhibited minimal toxicity against a p185-negative cell line (NIH3T3).

CONCLUSIONS

Selective and efficient killing of human lung adenocarcinoma cells can be achieved in vitro using c-erbB-2/p185-directed therapy.

摘要

背景

进行这些实验是为了确定使用抗p185-凝胶onin缀合物是否可以在肺腺癌中对p185进行治疗靶向。c-erbB-2/p185在高达三分之一的非小细胞肺癌中过表达。CALU-3是一种过表达p185的人肺腺癌细胞系。muMoAb-4D5是一种鼠抗p185单克隆免疫球蛋白G1。凝胶onin是一种有效的1型核糖体抑制蛋白。

方法

4D5和凝胶onin进行共价修饰并连接。通过SDS-聚丙烯酰胺凝胶电泳确认纯化。在用4D5-凝胶onin缀合物或对照物质（4D5、凝胶onin、未缀合的4D5 +凝胶onin或对照抗体MOPC-21）孵育后，使用CALU-3细胞摄取3H-胸苷来定量剂量依赖性细胞毒性。

结果

4D5-凝胶onin缀合物的50%抑制浓度为3.5×10^(-10) mol/L，但单独的4D5没有细胞毒性作用。凝胶onin和未缀合的4D5-凝胶onin混合物的细胞毒性是4D5-凝胶onin缀合物的十分之一（抑制浓度分别为6.5×10^(-9) mol/L和8.5×10^(-9) mol/L）。该缀合物对p185阴性细胞系（NIH3T3）表现出最小的毒性。