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第41卷第6期                           南京医科大学学报（自然科学版）
                  2021年6月                   Journal of Nanjing Medical University（Natural Sciences）     ·817 ·


               ·基础医学·

                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



                冯   恬 ，龚丹丹 ，王露瑶 ，王怡雯 ，周婷婷 ，朱                 进 ，张慧林 ，童       华  1*
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                南京医科大学附属妇产医院妇科，江苏 南京                 210004；东部战区疾病预防控制中心，江苏             南京 210002
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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⁃an⁃
                titumor 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。
               ［关键词］ c⁃Met；抗体药物偶联物；重组东亚钳蝎镇痛抗肿瘤多肽；卵巢癌
               ［中图分类号］ R737.31                   ［文献标志码］ A                      ［文章编号］ 1007⁃4368（2021）06⁃817⁃09
                doi：10.7655/NYDXBNS20210605


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

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                FENG Tian ，GONG Dandan ，WANG Luyao ，WANG Yiwen ，ZHOU Tingting ，ZHU Jin ，ZHANG Huilin ，TONG
                Hua 1*
                1 Department of Gynaecology，the Affiliated Obstetrics and Gynaecology Hospital of Nanjing Medical University，
                Nanjing 210004；Center of Eastern Theater for Disease Control and Prevention，Nanjing 210002，China
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               ［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.
               ［Key words］ c⁃Met；antibody⁃drug conjugate；recommbinant analgesic⁃antitumor peptide；ovarian cancer
                                                                              ［J Nanjing Med Univ，2021，41（06）：817⁃825］

               ［基金项目］ 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）
                ∗
                通信作者（Corresponding author），E⁃mail：zhl068@163.com；thua8@163.com
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