Suzuki Hiroyuki, Goto Nohara, Tanaka Tomohiro, Ouchida Tsunenori, Kaneko Mika K, Kato Yukinari
Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
Antibodies (Basel). 2023 Jul 4;12(3):45. doi: 10.3390/antib12030045.
Gastric cancer (GC) is the third leading cause of cancer-related deaths worldwide. GC with peritoneal metastasis exhibits a poor prognosis due to the lack of effective therapy. A comprehensive analysis of malignant ascites identified the genomic alterations and significant amplifications of cancer driver genes, including . CD44 and its splicing variants are overexpressed in tumors, and play crucial roles in the acquisition of invasiveness, stemness, and resistance to treatments. Therefore, the development of CD44-targeted monoclonal antibodies (mAbs) is important for GC diagnosis and therapy. In this study, we immunized mice with CD44v3-10-overexpressed PANC-1 cells and established several dozens of clones that produce anti-CD44v3-10 mAbs. One of the clones (CMab-94; IgG, kappa) recognized the variant-8-encoded region and peptide, indicating that CMab-94 is a specific mAb for CD44v8. Furthermore, CMab-94 could recognize CHO/CD44v3-10 cells, oral squamous cell carcinoma cell line (HSC-3), or GC cell lines (MKN45 and NUGC-4) in flow cytometric analyses. CMab-94 could detect the exogenous CD44v3-10 and endogenous CD44v8 in western blotting and stained the formalin-fixed paraffin-embedded gastric cancer cells. These results indicate that CMab-94 is useful for detecting CD44v8 in a variety of experimental methods and is expected to become usefully applied to GC diagnosis and therapy.
胃癌（GC）是全球癌症相关死亡的第三大主要原因。伴有腹膜转移的胃癌由于缺乏有效治疗，预后较差。对恶性腹水的综合分析确定了癌症驱动基因的基因组改变和显著扩增，包括。CD44及其剪接变体在肿瘤中过表达，并在侵袭性、干性和治疗抗性的获得中发挥关键作用。因此，开发靶向CD44的单克隆抗体（mAb）对胃癌的诊断和治疗很重要。在本研究中，我们用过表达CD44v3 - 10的PANC - 1细胞免疫小鼠，并建立了几十个产生抗CD44v3 - 10 mAb的克隆。其中一个克隆（CMab - 94；IgG，κ）识别变体8编码区域和肽，表明CMab - 94是CD44v8的特异性mAb。此外，在流式细胞术分析中，CMab - 94可以识别CHO/CD44v3 - 10细胞、口腔鳞状细胞癌细胞系（HSC - 3）或胃癌细胞系（MKN45和NUGC - 4）。CMab - 94可以在蛋白质印迹中检测外源性CD44v3 - 10和内源性CD44v8，并对福尔马林固定石蜡包埋的胃癌细胞进行染色。这些结果表明，CMab - 94在多种实验方法中可用于检测CD44v8，并有望有效地应用于胃癌的诊断和治疗。
