Bai Juan, Chen Ping, Zhou Qingxia, Tie Xiaojun, Xia Xiao, Wang Yan, Jin Ling
Department of Oncology, Wuzhong People's Hospital Affiliated to Ningxia Medical University, China.
The second Department of Oncology, Tumor Hospital, General Hospital of Ningxia Medical University, China.
Tissue Cell. 2025 Apr;93:102714. doi: 10.1016/j.tice.2024.102714. Epub 2024 Dec 31.
Macrophages in the tumor microenvironment (TME) regulated gastric cancer progression, but the mechanism of macrophage polarization in gastric cancer progression remained unclear. This study mainly explored the molecular mechanism of macrophage polarization in the tumor microenvironment and its impact on the progression of gastric cancer. KPNA2 and KPNB1 expressions in cancer tissues and adjacent non-cancerous tissues were quantified via RT-qPCR and western blot. A correlation analysis was conducted between KPNA2 and KPNB1 expressions, utilizing the GEPIA2 database to link them with macrophage polarization. KPNA2-KPNB1 interaction was investigated on STRING, verified by Co-IP and IF assays. Raw246.7 cells were transfected with KPNA2 overexpression with or without si-KPNB1 plasmids. Then, M1/M2 macrophage markers and the proportion of M2 macrophages were measured by RT-qPCR, western blot, and IF. Co-culturing transfected Raw246.7 with MFC cells showed gastric cancer cell proliferation, apoptosis, migration, and invasion via CCK-8, flow cytometry, and transwell assays. KPNA2 and KPNB1 in gastric cancer tissues were elevated, exhibiting a positive correlation between them. KPNA2 overexpression facilitated the differentiation of macrophages into M2 type. KPNA2 overexpression in macrophages co-cultured with MFC cells stimulated MFC cells proliferation, repressed apoptosis, and enhanced migration/invasion. The interaction between KPNA2 and KPNB1 was confirmed through Co-IP and IF assays. Si-KPNB1 reversed the effects of KPNA2 overexpression on macrophages and gastric cancer cells. KPNA2 promoted the M2 polarization of macrophages by upregulating KPNB1, thereby inducing the proliferation and metastasis of gastric cancer.
肿瘤微环境（TME）中的巨噬细胞调节胃癌进展，但其在胃癌进展中巨噬细胞极化的机制仍不清楚。本研究主要探讨肿瘤微环境中巨噬细胞极化的分子机制及其对胃癌进展的影响。通过RT-qPCR和蛋白质免疫印迹法对癌组织和癌旁非癌组织中的KPNA2和KPNB1表达进行定量。利用GEPIA2数据库将KPNA2和KPNB1表达与巨噬细胞极化联系起来，进行相关性分析。在STRING上研究KPNA2-KPNB1相互作用，并通过免疫共沉淀（Co-IP）和免疫荧光（IF）分析进行验证。用KPNA2过表达质粒转染Raw246.7细胞，转染时有无si-KPNB1质粒。然后，通过RT-qPCR、蛋白质免疫印迹法和免疫荧光法检测M1/M2巨噬细胞标志物及M2巨噬细胞比例。将转染后的Raw246.7与MFC细胞共培养，通过CCK-8、流式细胞术和Transwell实验检测胃癌细胞的增殖、凋亡、迁移和侵袭。胃癌组织中KPNA2和KPNB1升高，二者呈正相关。KPNA2过表达促进巨噬细胞向M2型分化。与MFC细胞共培养的巨噬细胞中KPNA2过表达刺激MFC细胞增殖，抑制凋亡，并增强迁移/侵袭。通过免疫共沉淀和免疫荧光分析证实了KPNA2和KPNB1之间的相互作用。Si-KPNB1逆转了KPNA2过表达对巨噬细胞和胃癌细胞的影响。KPNA2通过上调KPNB1促进巨噬细胞的M2极化，从而诱导胃癌的增殖和转移。
