Department of General Medicine, Cancer Hospital of Dalian University of Technology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, Shenyang 110042, China.
Molecular Oncology Department of Cancer Research Institution, The First Affiliated Hospital of China Medical University, Shenyang 110001, China.
J Immunol Res. 2023 Nov 6;2023:5532617. doi: 10.1155/2023/5532617. eCollection 2023.
Lung squamous cell carcinoma (LUSC) is associated with poor clinical prognosis and lacks available targeted agents. GPC3 is upregulated in LUSC. Our study aimed to explore the roles of GPC3 in LUSC and the antitumor effects of HLA-A2-restricted GPC3 antigenic peptide-sensitized dendritic cell (DC)-induced cytotoxic T lymphocytes (CTLs) on LUSC. LUSC cells with GPC3 knockdown and overexpression were built using lentivirus packaging, and cell viability, clone formation, apoptosis, cycle, migration, and invasion were determined. Western blotting was used to detect the expression of cell cycle-related proteins and PI3K-AKT pathway-associated proteins. Subsequently, HLA-A2-restricted GPC3 antigenic peptides were predicted and synthesized by bioinformatic databases, and DCs were induced and cultured . Finally, HLA-A2-restricted GPC3 antigenic peptide-modified DCs were co-cultured with T cells to generate specific CTLs, and the killing effects of different CTLs on LUSC cells were studied. A series of cell function experiments showed that GPC3 overexpression promoted the proliferation, migration, and invasion of LUSC cells, inhibited their apoptosis, increased the number of cells in S phase, and reduced the cells in G2/M phase. GPC3 knockdown downregulated cyclin A, c-Myc, and PI3K, upregulated E2F1, and decreased the pAKT/AKT level. Three HLA-A2-restricted GPC3 antigenic peptides were synthesized, with GPC3 FLAELAYDL and GPC3 FLIIQNAAV antigenic peptide-modified DCs inducing CTL production, and exhibiting strong targeted killing ability in LUSC cells at 80 : 1 multiplicity of infection. GPC3 may advance the onset and progression of LUSC, and GPC3 FLAELAYDL and GPC3 FLIIQNAAV antigenic peptide-loaded DC-induced CTLs have a superior killing ability against LUSC cells.
肺鳞状细胞癌（LUSC）与不良的临床预后相关，且缺乏可用的靶向药物。GPC3 在 LUSC 中上调。本研究旨在探讨 GPC3 在 LUSC 中的作用，以及 HLA-A2 限制性 GPC3 抗原肽致敏树突状细胞（DC）诱导的细胞毒性 T 淋巴细胞（CTL）对 LUSC 的抗肿瘤作用。使用慢病毒包装构建 GPC3 敲低和过表达的 LUSC 细胞，测定细胞活力、克隆形成、凋亡、细胞周期、迁移和侵袭。Western blot 用于检测细胞周期相关蛋白和 PI3K-AKT 通路相关蛋白的表达。随后，通过生物信息学数据库预测和合成 HLA-A2 限制性 GPC3 抗原肽，并诱导和培养 DC。最后，将 HLA-A2 限制性 GPC3 抗原肽修饰的 DC 与 T 细胞共培养，产生特异性 CTL，研究不同 CTL 对 LUSC 细胞的杀伤作用。一系列细胞功能实验表明，GPC3 过表达促进了 LUSC 细胞的增殖、迁移和侵袭，抑制了其凋亡，增加了 S 期细胞数量，减少了 G2/M 期细胞。GPC3 敲低下调了细胞周期蛋白 A、c-Myc 和 PI3K，上调了 E2F1，并降低了 pAKT/AKT 水平。合成了三个 HLA-A2 限制性 GPC3 抗原肽，GPC3 FLAELAYDL 和 GPC3 FLIIQNAAV 抗原肽修饰的 DC 诱导 CTL 产生，在 80：1 感染复数时对 LUSC 细胞具有较强的靶向杀伤能力。GPC3 可能促进 LUSC 的发生和发展，GPC3 FLAELAYDL 和 GPC3 FLIIQNAAV 抗原肽负载的 DC 诱导的 CTL 对 LUSC 细胞具有优越的杀伤能力。
