摘要
研究旨在探究miR-5589-5p对肝癌细胞凋亡和自噬的作用及机制。采用生物信息学分析miR-5589-5p、FOXK1在人肝细胞癌中的表达差异及其与肝癌患者预后的关系。采用miR-5589-5p mimics转染上调miR-5589-5p,检测miR-5589-5p及LC3、p62、Beclin-1、FOXK1、Bax、Bcl-2蛋白表达水平,检测细胞凋亡及自噬,双荧光素酶报告基因及RNA免疫共沉淀法检测miR-5589-5p与FOXK13′-UTR的结合。结果显示,miR-5589-5p在人肝细胞癌中低表达(P<0.05),miR-5589-5p表达水平越低肝癌预后越差(P<0.05)。FOXK1在人肝细胞癌中高表达(P<0.05),FOXK1表达水平越高肝癌预后越差(P<0.05)。miR-5589-5p过表达显著促进细胞凋亡和自噬(P<0.05)。miR-5589-5p能靶向结合FOXK1的3′-UTR。在miR-5589-5p mimics转染的基础上过表达FOXK1,可显著抑制miR-5589-5p mimics的上述作用(P<0.05)。综上,miR-5589-5p在肝癌中低表达,miR-5589-5p通过靶向FOXK1促进肝癌细胞凋亡和自噬。研究结果为进一步探讨miR-5589-5p作为肝细胞癌预测标志或治疗靶点提供了实验基础。
This study aimed to investigate the role and mechanism of miR-5589-5p on apoptosis and autophagy in hepatocellular carcinoma(HCC)cells.Bioinformatics analysis was employed to explore the differential expression of miR-5589-5p and FOXK1 in human HCC and their association with the prognosis of patients with HCC.miR-5589-5p mimics were used to upregulate miR-5589-5p levels.The levels of miR-5589-5p,protein expression levels of LC3,p62,Beclin-1,FOXK1,Bax,and Bcl-2 were detected.Cell apoptosis and autophagy were detected.Dual-luciferase reporter gene assay and RNA immunoprecipitation(RIP)were utilized to verify the binding of miR-5589-5p to the 3′-UTR of FOXK1.The results showed that miR-5589-5p was lowly expressed in human HCC(P<0.05),and a lower expression level of miR-5589-5p correlated with a poorer prognosis in HCC(P<0.05).In contrast,FOXK1 was highly expressed in HCC(P<0.05),and a higher expression level of FOXK1 was associated with a worse prognosis(P<0.05).Overexpression of miR-5589-5p significantly promoted cell apoptosis and autophagy.miR-5589-5p was confirmed to target the 3′-UTR of FOXK1.Overexpression of FOXK1 significantly inhibited the aforementioned effects of miR-5589-5p mimics(P<0.05).In conclusion,miR-5589-5p is lowly expressed in HCC and promotes apoptosis and autophagy in HCC cells by regulating FOXK1.This study lays an experimental foundation for further exploring the feasibility of miR-5589-5p as a predictive marker or therapeutic target for HCC.
作者
江策
刘昳
赵亚磊
常香荣
JIANG Ce;LIU Yi;ZHAO Yalei;CHANG Xiangrong(Department of Medical Laboratory,Shaanxi Provincial Rehabilitation Hospital,Xi′an 710065,China;Department of Traditional Chinese Medicine,the First Affiliated Hospital of Xi'an Jiaotong University,Xi′an 710065,China;Department of Infectious Diseases,the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710065,China;Department of Gastroenterology,Shaanxi Provincial Rehabilitation Hospital,Xi′an 710065,China)
出处
《生物技术进展》
2025年第6期1086-1093,共8页
Current Biotechnology
基金
陕西省自然科学基础研究计划项目(2022JQ-833)。
