Han Xiurui, Fu Weiwei, Sun Qinghua, Ning Jing, Zhang Jing, Matsas Silvio, de Melo Fabrício Freire, Zhang Hejun, Hao Xinyu, Meng Qiao, Gong Yueqing, Zheng Huiling, Zhang Jing, Ding Shigang
Department of Gastroenterology, Peking University Third Hospital, Beijing, China.
Beijing Key Laboratory for Helicobacter Pylori Infection and Upper Gastrointestinal Diseases (BZ0317), Beijing, China.
J Gastrointest Oncol. 2024 Aug 31;15(4):1431-1445. doi: 10.21037/jgo-24-466. Epub 2024 Aug 28.
CKLF-like MARVEL transmembrane domain-containing 4 (CMTM4) is involved in immune regulation and tumor progression; however, its role in gastric cancer (GC) remains unclear. This study explored the role and mechanism of CMTM4 in GC.
Immunohistochemistry was used to analyze CMTM4 expression in human gastric biopsied cells from patients with GC (N=23) or chronic superficial gastritis (N=23). To investigate the function of CMTM4 in GC cells, the gene was knocked down and overexpressed in human gastric adenocarcinoma cell line AGS. The gene was overexpressed in AGS cells and human gastric cell line SGC7901. Cell Counting Kit 8 (CCK-8) and cell clonogenic assays were used to analyze the proliferation of the GC cells. Flow cytometry was used to analyze the effects of CMTM4 on apoptosis and the cell cycle. Wound healing and transwell assays were used to analyze the migration and invasion of the gastric cells, respectively. The mechanism of CMTM4 in GC cells was explored using the tandem mass tags (TMTs) proteome and verified by western blot analysis.
CMTM4 expression was more downregulated in the human GC tissues than the gastritis tissues. CMTM4 overexpression significantly inhibited the proliferation, migration, and invasion of the GC cells, whereas CMTM4 knockdown enhanced gastric cell proliferation (P>0.05), migration (P>0.05), and invasion (P>0.05). Flow cytometry showed that CMTM4 promoted apoptosis and resulted in G1/S arrest in the GC cells. In addition, the proteome and western blot results showed that STAT1 was significantly upregulated, and the STAT1 signaling pathways were enriched in the GC cells overexpressing CMTM4.
Our results suggest that CMTM4 plays a tumor-suppressive role in GC and may affect the growth, migration, and invasion of GC cells through the STAT1 signaling pathway. CMTM4 might have potential value as a prognosis marker and potential therapeutic target for GC therapy.
含CKLF样MARVEL跨膜结构域4（CMTM4）参与免疫调节和肿瘤进展；然而，其在胃癌（GC）中的作用仍不清楚。本研究探讨CMTM4在GC中的作用及机制。
采用免疫组织化学分析CMTM4在GC患者（N = 23）或慢性浅表性胃炎患者（N = 23）的人胃活检细胞中的表达。为研究CMTM4在GC细胞中的功能，在人胃腺癌细胞系AGS中敲低并过表达该基因。在AGS细胞和人胃细胞系SGC7901中过表达该基因。使用细胞计数试剂盒8（CCK-8）和细胞克隆形成试验分析GC细胞的增殖。采用流式细胞术分析CMTM4对细胞凋亡和细胞周期的影响。分别使用伤口愈合试验和Transwell试验分析胃细胞的迁移和侵袭。使用串联质谱标签（TMT）蛋白质组学探索CMTM4在GC细胞中的机制，并通过蛋白质印迹分析进行验证。
CMTM4在人GC组织中的表达比胃炎组织中下调更明显。CMTM4过表达显著抑制GC细胞的增殖、迁移和侵袭，而CMTM4敲低增强胃细胞增殖（P>0.05）、迁移（P>0.05）和侵袭（P>0.05）。流式细胞术显示CMTM4促进凋亡并导致GC细胞G1/S期阻滞。此外，蛋白质组学和蛋白质印迹结果显示，在过表达CMTM4的GC细胞中，信号转导和转录激活因子1（STAT1）显著上调，且STAT1信号通路富集。
我们的结果表明，CMTM4在GC中发挥肿瘤抑制作用，并可能通过STAT1信号通路影响GC细胞的生长、迁移和侵袭。CMTM4可能作为GC治疗的预后标志物和潜在治疗靶点具有潜在价值。
