人源抗c⁃Met抗体与rAGAP偶联物对卵巢癌细胞生物学特性的影响
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江苏省重点研发计划专项(BE2018613);江苏省科教强卫工程青苗人才(QNRC2016104)


Effects of human anti⁃c⁃Met antibody conjugated with rAGAP on the biological properties of ovarian cancer cells
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    摘要:

    目的:制备人源抗c-Met的全分子抗体(c-Met-IgG1)及其与重组东亚钳蝎镇痛抗肿瘤多肽(recombinant analgesic-antitumor peptide,rAGAP)偶联的新型抗体(c-Met-IgG1-rAGAP),分析它们的生物学特性,检测其对卵巢癌细胞生物学行为的影响。方法:以本实验室保存的人源抗c-Met抗体基因为模板,构建c-Met-IgG1-rAGAP真核表达载体并转染真核细胞,表达并纯化抗体偶联物。通过亲和力检测、免疫荧光、流式细胞术等方法,评估c-Met- IgG1和c-Met-IgG1-rAGAP的生物学特性。通过CCK-8、划痕实验、Transwell侵袭实验检测抗体对卵巢癌细胞增殖、迁移和侵袭的影响。结果:成功制备出能够与c-Met蛋白特异性结合的c-Met-IgG1和c-Met-IgG1-rAGAP两种抗体;一系列实验证明了c-Met-IgG1和c-Met-IgG1-rAGAP均可抑制c-Met阳性的卵巢癌细胞HO8910的增殖、迁移、侵袭,且c-Met-IgG1-rAGAP对卵巢癌细胞的抑制作用要强于c-Met-IgG1,而在c-Met阴性的IOSE386细胞中并未观察到这些作用。结论:研究结果表明c-Met-IgG1-rAGAP可靶向作用于c-Met阳性的卵巢癌细胞并具有抗肿瘤活性,可为探索卵巢癌的分子靶向治疗奠定研究基础。

    Abstract:

    Objective:This study aims to prepare a human anti-c-Met full-molecule antibody(c-Met-IgG1)conjugated with recombinant analgesic-antitumor peptide(rAGAP) to analyze their biological characteristics and detect their effects on the biological behavior of ovarian cancer cells. Methods:Using the human anti-c-Met antibody gene stored in our laboratory as a template,construct the c-Met-IgG1-rAGAP eukaryotic expression vector was constructed and transfected it into eukaryotic cells to obtain the antibody conjugate. The immunological properties of c-Met-IgG1 and c-Met-IgG1-rAGAP were evaluated by methods such as affinity detection,immunofluorescence,and flow cytometry. CCK-8,scratch test,and Transwell invasion test were used to detect the effects of antibodies on the proliferation,migration and invasion of ovarian cancer cells. Results:Two antibodies,c-Met-IgG1 and c-Met-IgG1-rAGAP,which can specifically bind to c-Met protein,were successfully prepared;c-Met-IgG1 and c-Met-IgG1-rAGAP proved in a series of experiments can inhibit the proliferation,migration and invasion of the c-Met-positive ovarian cancer cell HO8910,and the inhibitory effect of c-Met-IgG1-rAGAP on ovarian cancer cells is higher than that of c-Met-IgG1,while these effects were not observed in the c-Met-negative IOSE386 cells. Conclusion:The research results show that c-Met-IgG1-rAGAP can target c-Met-positive ovarian cancer cells and has anti-tumor activity,which can lay a research foundation for exploring molecular targeted treatment of ovarian cancer.

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冯 恬,龚丹丹,王露瑶,王怡雯,周婷婷,朱 进,张慧林,童 华.人源抗c⁃Met抗体与rAGAP偶联物对卵巢癌细胞生物学特性的影响[J].南京医科大学学报(自然科学版),2021,(6):817-825

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  • 收稿日期:2021-01-07
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  • 在线发布日期: 2021-06-26
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