异种表皮生长因子受体胞外段的表达及其抗肿瘤效果研究

发布时间：2019-04-23 19:02

【摘要】：目的:构建异种EGFR蛋白疫苗并研究其抗肿瘤效果。方法:用PCR方法获得鸡EGFR胞外片段,与pGEX-4T-2载体连接,并在E.coli BL21(DE3)中表达鸡的EGFR胞外片段与GST的融合蛋白,融合蛋白经Ni2+亲和层析柱纯化、复性后,免疫小鼠,免疫3次后,接种小鼠Lewis细胞,观测肿瘤生长情况,ELISA检测小鼠血清中抗EGFR蛋白的抗体效价。结果:成功构建了EGFR胞外段基因的原核表达载体;SDS-PAGE显示成功表达了目的融合蛋白;融合蛋白免疫小鼠后,ELISA法检测到特异性抗EGFR抗体;实验组与对照组肿瘤体积的比较结果显示,融合蛋白疫苗能够在一定程度上抑制肿瘤的生长。结论:异种EGFR蛋白能够克服免疫耐受问题,诱导机体产生抗体,有一定的抗肿瘤效果,为后续的肿瘤疫苗研究打下了基础。
[Abstract]:Objective: to construct xenogeneic EGFR protein vaccine and study its antitumor effect. Methods: the extracellular fragment of chicken EGFR was obtained by PCR and ligated with pGEX-4T-2 vector. The fusion protein of chicken EGFR extracellular fragment and GST was expressed in E.coli BL21 (DE3). The fusion protein was purified by Ni2 affinity chromatography and renatured, then immunized mice. After 3 times of immunization, Lewis cells were inoculated into mice, the growth of tumor was observed, and the antibody titers of anti-EGFR protein in serum of mice were detected by ELISA. Results: the prokaryotic expression vector of the extracellular segment of EGFR was successfully constructed, and the fusion protein was successfully expressed by SDS-PAGE, and the specific anti-EGFR antibody was detected by ELISA after immunization with the fusion protein in mice. The results showed that the fusion protein vaccine could inhibit tumor growth to a certain extent. Conclusion: xenogeneic EGFR protein can overcome the problem of immune tolerance and induce the body to produce antibodies, which has a certain anti-tumor effect, which lays a foundation for further research on tumor vaccine.
【作者单位】：西北农林科技大学动物医学院;上海市肺科医院肺癌免疫研究室;沈阳农业大学畜牧兽医学院;
【基金】：上海市科委重大课题资助项目(03DZ19236)
【分类号】：R73-36

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