[鞘氨醇激酶-1通过靶向核因子-κB信号通路调控胃癌细胞的迁移和侵袭]
[Sphingosine kinase-1 regulates migration and invasion of gastric cancer cells targeting the nuclear factor-κB signaling pathway].
作者信息
Ling Q, Ji K, Chen J, Guan J, Wang R, Man W, Zhu B
机构信息
Department of Gastrointestinal Surgery, First Affiliated Hospital of Bengbu Medical University, Bengbu 233000, China.
Department of Surgery, Second Anhui Provincial People's Hospital, Hefei 230000, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Nov 20;44(11):2163-2171. doi: 10.12122/j.issn.1673-4254.2024.11.13.
OBJECTIVE
To investigate the role of sphingosine kinase-1 (SPHK1) in regulating migration and invasion of gastric cancer (GC) cells.
METHODS
TIMER2.0, GEPIA and HPA databases were used to investigate SPHK1 expression in GC, and its association with prognosis of the patients was analyzed using Kaplan-Meier Plotter database. In 40 clinical GC and adjacent tissue samples, SPHK1 and MKI67 expressions were detected with immunohistochemistry, Western blotting, and RT-qPCR. Gene enrichment pathway analysis was conducted to explore the biological functions of SPHK1. In HGC-27 and MGC-803 cells, the effects of lentivirus-mediated SPHK1 knockdown or overexpression on cell migration and invasion and expressions of key proteins in the nuclear factor-κB (NF-κB) signaling were evaluated using cell scratch test, Transwell assays and Western blotting. The changes in tumorigenic capacity of the transfected GC cells were evaluated in nude mice.
RESULTS
SPHK1 was highly expressed in GC tissues in negative correlation with overall survival, overall survival after progression, and relapse-free survival of the patients (all <0.001). In clinical GC samples, SPHK1 and MKI67 expressions showed a positive correlation (= 0.00049) and were both significantly up-regulated (<0.001). Gene enrichment pathway analysis suggested the involvement of SPHK1 in cell adhesion, migration, angiogenesis and the NF-κB pathway (<0.05). In the cell experiment, SPHK1 knockdown significantly decreased while SPHK1 overexpression enhanced migration and invasion abilities of the GC cells. SPHK1 positively regulated the expressions of phosphorylated P65 (P-P65), VEGFA and IL-17, and blocking the NF-κB pathway by PDTC significantly lowered migration and invasion ability of the cells. In nude mice, the GC cells with SPHK1 knockdown resulted in significantly reduced tumor size and mass, while the SPHK1-overexpressing cells showed enhanced tumorigenicity.
CONCLUSION
SPHK1 regulates migration and invasion of GC cells via the NF-κB signaling pathway and may serve as a potential diagnostic marker for GC progression.
目的
探讨鞘氨醇激酶-1（SPHK1）在调节胃癌（GC）细胞迁移和侵袭中的作用。
方法
利用TIMER2.0、GEPIA和HPA数据库研究GC中SPHK1的表达，并使用Kaplan-Meier Plotter数据库分析其与患者预后的关系。在40例临床GC及癌旁组织样本中，采用免疫组织化学、蛋白质免疫印迹法和逆转录定量聚合酶链反应检测SPHK1和MKI67的表达。进行基因富集通路分析以探索SPHK1的生物学功能。在HGC-27和MGC-803细胞中，使用细胞划痕试验、Transwell实验和蛋白质免疫印迹法评估慢病毒介导的SPHK1敲低或过表达对细胞迁移、侵袭以及核因子-κB（NF-κB）信号通路中关键蛋白表达的影响。在裸鼠中评估转染的GC细胞致瘤能力的变化。
结果
SPHK1在GC组织中高表达，与患者的总生存期、疾病进展后的总生存期和无复发生存期呈负相关（均<0.001）。在临床GC样本中，SPHK1和MKI67的表达呈正相关（=0.00049），且均显著上调（<0.001）。基因富集通路分析表明SPHK1参与细胞黏附、迁移、血管生成和NF-κB通路（<0.05）。在细胞实验中，SPHK1敲低显著降低而SPHK1过表达增强了GC细胞的迁移和侵袭能力。SPHK1正向调节磷酸化P65（P-P65）、血管内皮生长因子A（VEGFA）和白细胞介素-17（IL-17）的表达，用吡咯烷二硫代氨基甲酸盐（PDTC）阻断NF-κB通路显著降低细胞的迁移和侵袭能力。在裸鼠中，敲低SPHK1的GC细胞导致肿瘤大小和质量显著减小，而过表达SPHK1的细胞则显示出更强的致瘤性。
结论
SPHK1通过NF-κB信号通路调节GC细胞的迁移和侵袭，可能作为GC进展的潜在诊断标志物。
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