SLC35E4基因对人肝内胆管癌细胞增殖、迁移和凋亡的作用研究

Study on the Role of SLC35E4 Gene in Proliferation, Migration, and Apoptosis of Human Intrahepatic Cholangiocarcinoma Cells

摘要: 目的：探究溶质载体家族35，成员E4 (Solute Carrier Family 35, Member E4, SLC35E4)调控肝内胆管癌(intrahepatic cholangiocarcinoma, iCCA)体外细胞增殖、迁移及凋亡的作用研究。方法：利用TIMER在线数据库进行泛癌分析SLC35E4在多种癌症中的表达情况，并通过TCGA和GEO数据集进行验证；同时利用该数据库分析SLC35E4的诊断和预后价值；通过CCK-8、克隆形成、细胞划痕及Hoechst 33342凋亡试剂盒检测SLC35E4敲低对人肝胆管癌细胞系(RBE细胞系)增殖、迁移及凋亡情况。结果：SLC35E4在肿瘤组织中高表达。生存曲线证实了SLC35E4的高表达相关与预后不良相关，表明SLC35E4具有较好的预后价值。CCK8和克隆形成实验结果显示，敲低SLC35E4基因可以抑制肝内胆管癌细胞RBE的增殖(p < 0.05)。划痕实验结果显示，敲低SLC35E对RBE细胞的迁移具有抑制作用。相较于NC组，SLC35E4敲低后的RBE细胞中呈现明显的凋亡染色；WB结果显示，敲低SLC35E4后，凋亡通路中关键蛋白均有所增加。上述结果表明敲低SLC35E4后可促进肝内胆管癌细胞RBE的细胞凋亡。结论：SLC35E4作为iCCA一种潜在的生物标志物，通过体外实验证实了肝内胆管癌中敲低SLC35E4可抑制肿瘤的进展，可能为肝内胆管癌患者预后评估及新型靶向治疗提供新思路。

Abstract: Objective: Investigation on the role of Solute Carrier Family 35, Member E4 (SLC35E4) in regulating the in vitro cell proliferation, migration and apoptosis of intrahepatic cholangiocarcinoma (iCCA). Methods: Using the TIMER online database for pan-cancer analysis, the expression of SLC35E4 in various cancers was investigated, and verified through TCGA and GEO; at the same time, the diagnostic and prognostic value of SLC35E4 was analyzed using this database; the effects of SLC35E4 knockdown on the proliferation, migration and apoptosis of human hepatobiliary cancer cell lines (RBE cell line) were detected through CCK-8, clone formation, cell scratch, and Hoechst 33342 apoptosis kits. Results: SLC35E4 is highly expressed in tumor tissues. The survival curve confirmed that the high expression of SLC35E4 is associated with poor prognosis, indicating that SLC35E4 has good prognostic value. The results of CCK8 and clone formation experiments showed that knocking down the SLC35E4 gene can inhibit the proliferation of iCCA cells RBE (p < 0.05). The scratch experiment results showed that knocking down SLC35E inhibited the migration of RBE cells. Compared with the NC group, obvious apoptotic staining was observed in RBE cells after SLC35E4 knockdown; WB results showed that after knocking down SLC35E4, the key proteins in the apoptotic pathway all increased. These results indicate that knocking down SLC35E4 can promote the apoptosis of iCCA cells RBE. Conclusion: SLC35E4, as a potential biomarker for iCCA, has been confirmed through in vitro experiments that knocking down SLC35E4 in iCCA can inhibit tumor progression. This finding may provide new ideas for prognosis assessment and novel targeted therapy for iCCA patients.