CEACAM6通过调控上皮间质转化抑制鼻咽癌细胞的增殖和迁移

doi:10.12122/j.issn.1673-4254.2025.03.14

摘要/Abstract

摘要：

目的明确CEACAM6在鼻咽癌中的表达，探索CEACAM6对鼻咽癌细胞增殖和迁移的影响以及潜在的作用机制。方法通过GEO数据库分析CEACAM6在鼻咽癌中表达情况。TCGA数据库分析CEACAM6在头颈部癌中表达情况以及与生存预期的关系。免疫组织化学技术检测CEACAM6在鼻咽癌组织中的表达情况。构建CEACAM6过表达细胞模型，分为control组（空载）和Lv-CEACAM6组（过表达）；CEACAM6敲低细胞模型，分为control组（空载）、sh#1组（敲低）和sh#2组（敲低）。通过CCK-8和Edu染色检测CEACAM6对鼻咽癌细胞增殖的影响。伤口愈合和Transwell检测CEACAM6对鼻咽癌细胞侵袭迁移的影响。鬼笔环肽染色检测CEACAM6对鼻咽癌细胞骨架排列的影响。Western blotting检测CEACAM6对鼻咽癌细胞中FN1、ITGA5、ITGB1、N-cad、Vim和E-cad的蛋白表达影响以及FN1过表达对CEACAM6过表达的鼻咽癌细胞中ITGA5和ITGB1蛋白表达的影响。结果 GEO数据库分析表明CEACAM6在鼻咽癌中低表达，TCGA数据库分析表明CEACAM6在头颈部癌中低表达，CEACAM6低表达与预后不良有关。免疫组织化学技术检测结果显示CEACAM6在鼻咽癌组织中低表达。CCK-8和Edu检测结果显示CEACAM6可抑制鼻咽癌细胞的增殖能力（P<0.05）。伤口愈合和Transwell检测结果显示CEACAM6可抑制鼻咽癌细胞侵袭和迁移能力（P<0.05）。鬼笔环肽染色结果显示上调CEACAM6表达后肌动蛋白荧光表达降低。Westen blotting检测结果表明上调CEACAM6表达后FN1、ITGA5、ITGB1下调，E-cad上调，N-cad和Vim下调（P<0.05），FN1过表达可缓解CEACAM6对ITGA5和ITGB1表达的抑制作用（P<0.05）。结论 CEACAM6可能通过调控FN1、ITGA5、ITGB1蛋白表达影响上皮间质转化从而抑制鼻咽癌的侵袭和迁移。

关键词: CEACAM6, 鼻咽癌, 迁移, 上皮间质转化

Abstract:

Objective To investigate CEACAM6 expression in nasopharyngeal carcinoma (NPC) and its regulatory effects on tumor cell proliferation, migration, and epithelial-mesenchymal transition (EMT). Methods CEACAM6 expression in NPC was analyzed using GEO datasets and validated by immunohistochemistry in NPC tissues and by Western blotting and RT-qPCR in NPC cell lines (HNE1, C666-1, HK1, 5-8F and CNE2Z) and normal nasopharyngeal epithelial NP69 cells. In the NPC cell lines, the effects of lentivirus-mediated CEACAM6 overexpression and knockdown on cell proliferation, migration, invasion and cytoskeletal structures were evaluated using CCK-8 assay, Edu staining, wound healing assay, Transwell assay, and phalloidin staining. Western blotting was performed to determine the expressions of EMT-related proteins (FN1, ITGA5, ITGB1, E-cadherin, N-cadherin and vimentin) in the NPC cells and the effect of FN1 overexpression on ITGA5 and ITGB1 protein expressions. Results Analysis of the data from the GEO datasets suggested that CEACAM6 was significantly downregulated in NPC, which was associated with poor patient prognosis. Immunohistochemistry also showed low expressions of CEACAM6 in clinical NPC tissues (P<0.05). In NPC cells, CEACAM6 overexpression significantly suppressed cell proliferation, migration and invasion and reduced the fluorescence intensity of actin. CEACAM6 overexpression also resulted in significant downregulation of FN1, ITGA5, ITGB1, N-cadherin and vimentin expressions and upregulation of E-cadherin expression, and FN1 overexpression obviously attenuated the inhibitory effect of CEACAM6 overexpression on ITGA5 and ITGB1 expressions. Conclusion CEACAM6 inhibits NPC cell migration and invasion by inhibiting EMT via regulating FN1, ITGA5 and ITGB1 expressions.

Key words: CEACAM6, nasopharyngeal carcinoma, migration, epithelial-mesenchymal transition

陶露, 韦卓利, 王月月, 项平. CEACAM6通过调控上皮间质转化抑制鼻咽癌细胞的增殖和迁移[J]. 南方医科大学学报, 2025, 45(3): 566-576.

Lu TAO, Zhuoli WEI, Yueyue WANG, Ping XIANG. CEACAM6 inhibits proliferation and migration of nasopharyngeal carcinoma cells by suppressing epithelial-mesenchymal transition[J]. Journal of Southern Medical University, 2025, 45(3): 566-576.

图/表 7

图1 CEACAM6 在肿瘤中的表达

Fig.1 CEACAM6 expression in nasopharyngeal carcinoma (NPC). A: GEO database analysis of CEACAM6 expression in NPC. B: Clustering dendrogram of co-expression network modules. C: Heatmap of correlation between gene modules and clinical characteristics. D: Venn diagram between GEO-diff and GEO-green, with a total of 107 crossed genes as differentially co-expressed genes. E: PPI network of 107 genes. F, G: Expression analysis and prognosis analysis of CEACAM6 in HNSC in the TCGA database. *P<0.05 vs Tumor.

图2 CEACAM6在鼻咽癌组织和细胞中低表达

Fig.2 CEACAM6 is lowly expressed in NPC tissues and cells. A: Immunohistochemistry for detecting CEACAM6 expression in NPC tissues. B, C: Western blotting and fluorescence quantitative PCR for detecting protein and mRNA levels of CEACAM6 in NPC cells. *P<0.05, **P<0.01 vs NP69.

图3 CEACAM6 对鼻咽癌细胞增殖的影响

Fig.3 Effect of CEACAM6 on NPC cell proliferation. A: CCK-8 assay for analyzing the effect of CEACAM6 on viability of HNE1 and C666-1 cells. B, C: Edu staining for detecting the effect of CEACAM6 on proliferation of HNE1 and C666-1 cells. *P<0.05, **P<0.01 vs control.

图4 CEACAM6 对鼻咽癌细胞侵袭和迁移的影响

Fig.4 Effect of CEACAM6 on invasion and migration of NPC cells. A, B: Wound healing assay for assessing the effect of CEACAM6 on migration of HNE1 and C666-1 cells. C-E: Transwell assay for assessing the effect of CEACAM6 on invasion and migration of HNE1 and C666-1 cells (×200). **P<0.01 vs Control.

图5 CEACAM6 调控 FN1/ITGA5/ITGB1蛋白表达

Fig.5 CEACAM6 regulates FN1/ITGA5/ITGB1 protein expressions. A: GSEA enrichment analysis. B, C: PPI and Cytoscape protein interactions. D, E: GEO database analysis of the expressions of CEACAM6, FN1 and ITGB1 in NPC and the correlation of CEACAM6 with FN1 and ITGB1. F-H: Western blotting for detecting the effect of CEACAM6 on FN1, ITGA5 and ITGB1 proteins in HNE1 and C666-1 cells. *P<0.05, **P<0.01 vs Control; #P<0.05 vs Lv-CEACAM6.

图6 FN1过表达对CEACAM6过表达的HNE1细胞增殖、侵袭和迁移的影响

Fig.6 Effects of FN1 overexpression on proliferation, invasion and migration of HNE1 cells with CEACAM6 overexpression. A: CCK-8 assay for assessing the effect of FN1 overexpression on viability of HNE1 cells with CEACAM6 overexpression. B: Edu staining for assessing the effect of FN1 overexpression on proliferation of HNE1 cells overexpressing CEACAM6. C: Wound healing assay for assessing the effect of FN1 overexpression on migration of HNE1 cells overexpressing CEACAM6. D-F: Transwell assay for assessing the effect of FN1 overexpression on invasion and migration of HNE1 cells overexpressing CEACAM6 (×200). *P<0.05, **P<0.01 vs Lv-CEACAM6.

图7 CEACAM6 调控 EMT 通路

Fig.7 CEACAM6 regulates the EMT pathway. A: TRITC-labeled phalloidin staining for analyzing the effect of CEACAM6 on cytoskeleton of HNE1 and C666-1 cells. B, C: Western blotting for detecting the effect of CEACAM6 on EMT-related proteins E-cadherin, N-cadherin, and vimentin in HNE1 and C666-1 cells. *P<0.05; **P<0.01 vs control.

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