摘要
目的:探讨长链非编码RNA羰基还原酶3反义RNA1(CBR3-AS1)调节微小RNA-145-5p(miR-145-5p)/S期激酶关联蛋白1(SKP1)轴对结肠癌细胞生物学行为的影响。方法:将结肠癌细胞系SW480细胞分为对照组、si-NC组、si-CBR3-AS1组、si-CBR3-AS1+anti-miR-NC组和si-CBR3-AS1+anti-miR-145-5p组;RT-qPCR检测细胞中CBR3-AS1、miR-145-5p、SKP1 mRNA的表达;CCK-8实验检测24、48、72 h的细胞增殖;Transwell实验检测细胞迁移和侵袭;流式细胞术检测细胞凋亡;免疫印迹检测细胞中SKP1、B淋巴细胞瘤-2相关蛋白(BAX)、B淋巴细胞瘤-2(Bcl-2)蛋白表达;双荧光素酶实验验证miR-145-5p与CBR3-AS1、SKP1 mRNA的关系。结果:与对照组、si-NC组比较,si-CBR3-AS1组SW180细胞中miR-145-5p、细胞凋亡率、BAX蛋白表达升高,SKP1 mRNA和CBR3-AS1表达、OD450值(24、48、72 h)、迁移和侵袭细胞数、Bcl-2蛋白表达降低。下调miR-145-5p减弱了低表达CBR3-AS1对SW480细胞增殖、迁移和侵袭的抑制作用,并抑制细胞凋亡;CBR3-AS1靶向负调控miR-145-5p表达,miR-145-5p靶向负调控SKP1 mRNA表达。结论:CBR3-AS1/miR-145-5p/SKP1轴可抑制SW480细胞增殖、迁移和侵袭,并促进其凋亡。
Objective:To investigate the impacts of long non-coding RNA carbonyl reductase 3-AS1(CBR3-AS1)on the malignant biological behavior of colon cancer cells by regulating microRNA-145-5p(miR-145-5p)/S-phase kinase-associated protein 1(SKP1)axis.Methods:Colon cancer cell line SW480 was divided into five groups:control group,si-NC group,si-CBR3-AS1 group,si-CBR3-AS1+anti miR-NC group,and si-CBR3-AS1+anti miR-145-5p group.RT-qPCR was applied to detect the expression of CBR3-AS1,miR-145-5p,and SKP1 mRNA in cells.CCK-8 assay was applied to detect cell proliferation.Transwell assay was utilized to detect cell migration and invasion abilities,and the flow cytometry was employed to detect the level of cell apoptosis.Western blotting was adopted to detect the expression levels of SKP1,B-cell lymphoma protein 2(Bcl-2)-associated X(Bax),and B-cell lymphoma protein 2(Bcl-2)in cancer cells.Dual-luciferase reporter assay was applied to verify the interaction between miR-145-5p and CBR3-AS1,as well as between miR-145-5p and SKP1 mRNA.Results:Compared with the control group and si-NC group,the miR-145-5p,apoptosis rate,and the expression of Bax protein in SW180 cells in the si-CBR3-AS1 group were increased,while the expression of SKP1 mRNA,CBR3-AS1,OD450 values(24,48,72 h),numbers of migrating and invading cells,and the expression of Bcl-2 protein were decreased.Down-regulation of miR-145-5p weakened the inhibitory effects of low-expressed CBR3-AS1 on the proliferation,migration,and invasion of SW480 cells,and also inhibited cell apoptosis.CBR3-AS1 targeted and negatively regulated the miR-145-5p expression,while miR-145-5p targeted and negatively regulated the SKP1 mRNA expression.Conclusion:The CBR3-AS1/miR-145-5p/SKP1 axis can inhibit proliferation,migration,and invasion of SW480 cells,and promote apoptosis in SW480 cells.
作者
程佩宇
申旭海
梁正子
Cheng Peiyu;Shen Xuhai;Liang Zhengzi(Department of Surgery,Beijing Fengtai Hospital of Integrated Traditional and Western Medicine,Beijing 100072;Department of Proctology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China)
出处
《解剖学杂志》
2025年第2期149-153,173,共6页
Chinese Journal of Anatomy
