Okita Kouki, Imai Kazuki, Kato Kazunori, Sugiura Reiko, Endo Yuichi, Masuko Kazue, Tomioka Yoshihisa, Masuko Takashi
Cell Biology Laboratory, School of Pharmacy, Kindai University, Higashiosaka-shi, Osaka, Japan; Laboratory of Molecular Pharmacogenomics, Faculty of Pharmacy, Kindai University, Japan; Production and Manufacturing, Carna Biosciences, Inc., Minatojima-Minamimachi, Chuo-ku, Kobe, 650-0047, Japan.
Cell Biology Laboratory, School of Pharmacy, Kindai University, Higashiosaka-shi, Osaka, Japan.
Biochem Biophys Res Commun. 2021 Oct 22;576:59-65. doi: 10.1016/j.bbrc.2021.08.091. Epub 2021 Aug 31.
HER1-and HER2-targeted drugs are effective in cancer therapy, especially against lung, breast and colon malignancies; however, resistance of cancer cells to HER1-and HER2-targeted therapies is becoming a serious problem. The avidity/affinity constant (K) and growth inhibitory effect of anti-HER3 rat monoclonal antibodies (mAb, Ab1∼Ab6) in the presence of therapeutic mAb or low-molecular-weight inhibitors against HER family proteins were analyzed by flow cytometry-based Scatchard plots (Splot) and cell proliferation assay. The K of Ab3 and Ab6, but not Ab1 or Ab4, split into dual (high and low) modes of K, and Ab6 exhibited greater anti-proliferative effects against LS-174T colon cancer cells in the presence of Pertuzumab (anti-HER2 mAb). A high K by Ab6 and Ab6-mediated increased growth inhibition were observed against NCI-H1838 lung or BT474 breast cancer cells, respectively, in the presence of Panitumumab (anti-HER1 mAb) or Perutuzumab. A high K by Ab6 and Ab6-mediated increased anti-proliferative effects against NCI-H1838 or BT474 were also respectively observed in the presence of Erlotinib (HER1 inhibitor) or Lapatinib (HER1/HER2 inhibitor). In HER1-knockout (KO) NCI-H1838, the reactivity and K of Ab4 increased compared with in parent NCI-H1838. In HER1-KO or HER3-KO SW1116 colon cancer cells, dual modes of K with Pertuzumab were noted, and the combination Ab6 and Pertuzumab promoted growth inhibition of HER1-KO, but not of parent SW1116.
针对HER1和HER2的药物在癌症治疗中有效，尤其是对肺癌、乳腺癌和结肠恶性肿瘤；然而，癌细胞对HER1和HER2靶向治疗的耐药性正成为一个严重问题。通过基于流式细胞术的Scatchard图（Splot）和细胞增殖测定，分析了抗HER3大鼠单克隆抗体（mAb，Ab1∼Ab6）在治疗性mAb或针对HER家族蛋白的低分子量抑制剂存在下的亲和力常数（K）和生长抑制作用。Ab3和Ab6的K值（而非Ab1或Ab4）分为双（高和低）模式的K，并且在帕妥珠单抗（抗HER2 mAb）存在下，Ab6对LS-174T结肠癌细胞表现出更大的抗增殖作用。在西妥昔单抗（抗HER1 mAb）或帕妥珠单抗存在下，分别观察到Ab6对NCI-H1838肺癌细胞或BT474乳腺癌细胞具有高K值以及Ab6介导的生长抑制增加。在厄洛替尼（HER1抑制剂）或拉帕替尼（HER1/HER2抑制剂）存在下，也分别观察到Ab6对NCI-H1838或BT474具有高K值以及Ab6介导的抗增殖作用增加。在HER1基因敲除（KO）的NCI-H1838细胞中，与亲本NCI-H1838相比，Ab4的反应性和K值增加。在HER1-KO或HER3-KO的SW1116结肠癌细胞中，注意到与帕妥珠单抗存在双模式的K值，并且Ab6与帕妥珠单抗联合促进了HER1-KO细胞的生长抑制，但对亲本SW1116细胞无此作用。
