LncRNA DSCAM-AS1调节miR-431-5p/SOX9轴对肝癌活性和化疗耐药性的影响  

Effects of LncRNA DSCAM-AS1 on liver cancer activity and chemotherapy resistance by regulating the miR-431-5p/SOX9 axis

作　　者：洪帆 王富华 谭一清 Hong Fan;Wang Fuhua;Tan Yiqing(Department of Interventional Radiology,Wuhan Third Hospital,Wuhan 430060,China)

机构地区：[1]武汉市第三医院放射介入科,武汉430060

出　　处：《解剖学杂志》2025年第1期39-47,72,共10页Chinese Journal of Anatomy

摘　　要：目的:探究长链非编码RNA(LncRNA)唐氏综合征细胞黏附分子反义1(DSCAM-AS1)调节miR-431-5p/性别决定基因盒9(SOX9)轴对肝癌活性和化疗耐药性的影响。方法:qRT-PCR检测肝癌组织、肝癌细胞及HepG2耐药细胞HepG2/CDDP中DSCAM-AS1、miR-431-5p、SOX9 mRNA表达,免疫印迹检测其多药耐药相关蛋白1(MRP1)、增殖细胞核抗原(PCNA)、Bcl-2、Bax、SOX9蛋白表达。MTT法检测各组HepG2和HepG2/CDDP细胞活力,流式细胞术检测细胞凋亡。生物信息学、双荧光素酶实验、pull down实验、RIP实验验证DSCAMAS1、SOX9与miR-431-5p靶向关系。结果:DSCAM-AS1、SOX9在肝癌组织及细胞中高表达,miR-431-5p低表达;与HepG2细胞比较,HepG2/CDDP细胞中DSCAM-AS1、SOX9表达、细胞活力显著升高,miR-431-5p表达降低;沉默DSCAM-AS1抑制HepG2和HepG2/CDDP细胞增殖,促进凋亡,降低HepG2/CDDP细胞耐药性;抑制miR-431-5p减弱沉默DSCAM-AS1对HepG2和HepG2/CDDP细胞增殖的抑制、凋亡的促进作用,促进HepG2/CDDP细胞耐药性;生物信息学、双荧光素酶实验、pull down实验、RIP实验证实DSCAM-AS1、SOX9与miR-431-5p的靶向关系。结论:沉默DSCAM-AS1可能通过调节miR-431-5p/SOX9轴抑制HepG2细胞增殖,促进凋亡,降低HepG2/CDDP细胞耐药性。Objective:To investigate the effect of long non-coding RNA(LncRNA)Down syndrome cell adhesion molecule antisense 1(DSCAM-AS1)on liver cancer activity and chemotherapy resistance by regulating the miR-431-5p/sex determining gene box 9(SOX9)axis.Methods:qRT-PCR was applied to detect the expression levels of DSCAM-AS1,miR-431-5p,and SOX9 mRNA in liver cancer tissue,liver cancer cells,and HepG2/CDDP resistant cells.Western blotting was utilized to detect the expression of multidrug resistance associated protein 1(MRP1),proliferating cell nuclear antigen(PCNA),Bcl-2,Bax,SOX9.Functional assays included MTT for cell viability,flow cytometry for apoptosis.Bioinformatics,dual luciferase assay,pull down assay,and RIP assay were applied to validate the targeting relationship between DSCAM-AS1,SOX9,and miR-431-5p.Results:DSCAM-AS1 and SOX9 were highly expressed in liver cancer tissues and cells,while miR-431-5p was low expressed.compared with HepG2 cells,the expression of DSCAM-AS1 and SOX9,and the cell viability in HepG2/CDDP cells were significantly increased,while the expression of miR-431-5p was reduced.Silencing DSCAM-AS1 inhibited the proliferation of HepG2 and HepG2/CDDP cells,induced apoptosis,and reduced drug resistance of HepG2/CDDP cells,while inhibiting miR-431-5p weakened the inhibitory effect of silencing DSCAM-AS1 on HepG2 and HepG2/CDDP cell proliferation,promoted apoptosis,and promoted drug resistance in HepG2/CDDP cells.Bioinformatics,dual luciferase assay,pull down assay,and RIP assay confirmed the targeting relationship between DSCAM-AS1,SOX9,and miR-431-5p.Conclusion:Silencing DSCAMAS1 may inhibit HepG2 cell proliferation,promote apoptosis,and reduce drug resistance in HepG2/CDDP cells by regulating the miR-431-5p/SOX9 axis.

关 键 词：肝癌 化疗耐药性 长链非编码RNA唐氏综合征细胞黏附分子反义1 miRNA-431-5p 性别决定基因盒9 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]