南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (9): 1613-1621.doi: 10.12122/j.issn.1673-4254.2023.09.20

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SLC12A8通过JAK/STAT途径促进膀胱癌细胞的增殖、侵袭与迁移并触发上皮间充质转化

张晓林，吴浩松，汪 盛   

  1. 蚌埠医学院第一附属医院泌尿外科，安徽 蚌埠 233004
  • 出版日期:2023-09-20 发布日期:2023-09-28

SLC12A8 promotes proliferation, invasiveness, migration and epithelial-mesenchymal transition of bladder cancer cells by activating JAK/STAT singaling

ZHANG Xiaolin, WU Haosong, WANG Sheng   

  1. Department of Urology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China
  • Online:2023-09-20 Published:2023-09-28

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摘要：

目的 探究溶质载体家族12成员8（SLC12A8）在膀胱癌中的表达及作用机制。方法 利用TCGA数据库分析SLC12A8在膀胱癌中的表达、预后及其与临床病理特征关系。细胞通过瞬时转染siRNA，将实验分为siNC组和siSLC12A8组，各组终浓度为50 nmol/L。利用qRT-PCR检测干扰效率，CCK-8、Transwell及划痕实验检测各组膀胱癌细胞增殖、侵袭与迁移能力。GSEA对通路富集分析；分析SLC12A8与EMT标志物相关性。Western blot检测通路蛋白及EMT相关蛋白表达。通过应用Colivelin将实验分为siSLC12A8+NC组和siSLC12A8+Colivelin组，药物浓度为0.5 μmol/L，利用cck-8及Transwell检测各组细胞生物学行为。结果 SLC12A8在膀胱癌中高表达（P<0.05）、与不良预后及病理分期相关（P<0.05），并可作为独立预后因素。CCK-8、Transwell及划痕实验结果表明，与siNC组相比，siSLC12A8组细胞增殖、侵袭与迁移能力降低（P<0.05）。GSEA表明富集在JAK/STAT信号通路上（P=0.008）。相关性分析表明，SLC12A8与E-cadherin负相关（r=-0.183，P<0.001），而与N-cadherin正相关（r=0.302，P<0.001）、vimentin正相关（r=0.342，P<0.001）。Western blot结果显示，与siNC组相比，siSLC12A8组p-Jak2、p-Stat3蛋白水平降低，E-cadherin表达增加、N-cadherin和vimentin蛋白表达水平下降。应用Colivelin后，与siSLC12A8+NC组相比，siSLC12A8+Colivelin组细胞增殖、侵袭与迁移能力增加（P<0.05）。结论 SLC12A8通过激活JAK/STAT信号通路促进膀胱癌进展，且与不良预后密切相关。

关键词:

Abstract: Objective To investigate the role of solute carrier family 12 member A8 (SLC12A8) in regulation of biological behaviors of bladder cancer and the mechanism mediating its effect. Methods The TCGA database was used to analyze SLC12A8 expression in bladder cancer and is correlation with prognosis and clinicopathological characteristics of the patients. In different bladder cancer cell lines, the effects of transient transfection with SLC12A8 siRNA on cell proliferation, invasion and migration ability were examined using CCK-8 assay, Transwell assay and scratch experiment. Gene set enrichment analysis (GSEA) was carried out to analyze pathway enrichment. The correlation of SLC12A8 with the expressions of epithelial-mesenchymal transition (EMT) markers was analyzed using Western blotting. The effect of colivelin on biological behaviors of the cells with SLC12A8 knockdown was assessed using CCK-8 and Transwell assays. Results SLC12A8 was highly expressed in bladder cancer (P<0.05) and associated with a poor prognosis and advanced pathological stages of the patients (P<0.05), and could serve as an independent prognostic factor. The bladder cancer cell lines with SLC12A8 knockdown showed significantly attenuated proliferation, invasion and migration capacities (P<0.05). GSEA identified significant gene enrichment in the JAK/STAT signaling pathway (P=0.008). Correlation analysis showed that SLC12A8 expression was negatively correlated with E-cadherin expression (r=- 0.183, P<0.001) but positively with N-cadherin (r=0.302, P<0.001) and vimentin (r=0.342, P<0.001) expressions. The bladder cancer cells with SLC12A8 knockdown showed significantly decreased expressions of p-Jak2, p-Stat3, N-cadherin and vimentin proteins with an increased expression of E-cadherin. Treatment with colivelin effectively enhanced proliferation, invasion and migration capacities of the bladder cancer cells with SLC12A8 knockdown (P<0.05). Conclusion SLC12A8 promotes bladder cancer progression by activating the JAK/STAT signaling pathway and its high expression is closely associated with a poor prognosis of the patients.

Key words: SLC12A8; bladder cancer; JAK/STAT; proliferation; epithelial-mesenchymal transition