Potent inhibition of human gastric cancer by HER2-directed induction of apoptosis with anti-HER2 antibody and caspase-3 fusion protein
Potent inhibition of human gastric cancer by HER2-directed induction of apoptosis with anti-HER2 antibody and caspase-3 fusion protein
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DOI：
10.1136/gut.2008.155226
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发表时间：
2009-11
期刊：
影响因子：
24.5
通讯作者：
De-xin Zhang;P. Zhao;L. Xia;Li-Li Liu-Li;Jie Liang;Hui-hong Zhai;Hong-bo Zhang;Xue-Gang Guo;Kai‐chun Wu;Yan-ming Xu;L. Jia;A. Yang;Si-Yi Chen;D. Fan
中科院分区：
文献类型：
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作者：
De-xin Zhang;P. Zhao;L. Xia;Li-Li Liu-Li;Jie Liang;Hui-hong Zhai;Hong-bo Zhang;Xue-Gang Guo;Kai‐chun Wu;Yan-ming Xu;L. Jia;A. Yang;Si-Yi Chen;D. Fan
背景和目的HER 2是一种癌基因，在10-40%的胃癌中过度表达。本研究的目的是研究由抗HER 2 sFv和组成型活性caspase-3组成的融合蛋白是否能够诱导HER 2表达的人胃癌细胞凋亡并阻断人胃癌裸鼠移植瘤的生长。方法将含有分泌信号、抗HER 2单链抗体片段、假单胞菌外毒素A易位结构域和组成型活性caspase-3分子的重组质粒pcDNA 3. 1-HER-PE-CP 3稳定转染NIH 3 T3细胞，在体外和体内诱导人胃癌细胞凋亡。免疫荧光染色和Western blotting检测重组蛋白HER-PE-CP 3的表达。流式细胞术和TUNEL法检测细胞凋亡。结果HER-PE-CP 3/NIH 3 T3与人胃癌细胞共培养后，HER 2表达的人胃癌细胞发生HER-PE-CP 3内化和凋亡，而HER 2阴性的人胃癌细胞则无此现象。HER-PE-CP 3/NIH 3 T3对人胃癌裸鼠移植瘤有明显的抑制作用，荷瘤小鼠的生存期明显延长。在接受HER-PE-CP 3/NIH 3 T3的小鼠的异种移植物中检测到比对照小鼠显著更多的凋亡细胞。结论HER-PE-CP 3嵌合分子可诱导HER 2阳性胃癌细胞选择性凋亡，并具有较强的生长抑制作用，为HER 2靶向治疗胃癌提供了新的思路。
Background and aims HER2, an oncogene, has been found to be over-expressed in 10–40% of human gastric carcinomas. The aims of this study were to investigate if a fusion protein consisting of anti-HER2 sFv and constitutively active caspase-3 was capable of inducing apoptosis in HER2-expressing human gastric cancer cells and blocking the growth of human gastric cancer xenografts in nude mice. Methods NIH3T3 cells stably transduced with the pcDNA3.1-HER-PE-CP3 recombinant plasmid containing a secretion signal, a single-chain anti-HER2 monoclonal antibody fragment, a Pseudomonas exotoxin A translocation domain and a constitutively active caspase-3 molecule were used to induce apoptosis in human gastric cancer cells both in vitro and in vivo. Immunofluorescence staining and western blotting were used to examine the expression of the recombinant protein HER-PE-CP3. Apoptosis was determined by flow cytometry and TUNEL assay. Results Co-cultivation of HER-PE-CP3/ NIH3T3 with human gastric cancer cells led to internalisation of HER-PE-CP3 and apoptosis in HER2-expressing human gastric cancer cells but not in HER2-negative cancer cells. Inoculation of HER-PE-CP3/NIH3T3 in nude mice resulted in potent inhibition of human gastric cancer xenografts and much prolonged survival time of the tumour-bearing mice compared with the control. Significantly more apoptotic cells were detected in xenografts in mice receiving HER-PE-CP3/NIH3T3 than in control mice. Conclusions The HER-PE-CP3 chimeric molecule could induce selective apoptosis and potent growth inhibition of HER2-positive human gastric cancer cells and might represent a novel HER2-directed treatment option for human gastric cancer.