长链非编码RNA F-box富含亮氨酸的重复蛋白19反义RNA 1调节微小RNA-342-3p/自噬相关蛋白10轴对结直肠癌细胞增殖、迁移和奥沙利铂耐药性的影响实验研究  

Effects of lncRNA FBXL19-AS1 on proliferation,migration,and oxaliplatin resistance of colorectal cancer cells by regulating miR-342-3p/ATG10 axis

作　　者：屠佳佳 王利云 孙波 王凡荣 冯辉 TU Jiajia;WANG Liyun;SUN Bo;WANG Fanrong;FENG Hui(Shuyang Hospital Affiliated to Nanjing University of Traditional Chinese Medicine,Shuyang 223600,China)

机构地区：[1]南京中医药大学附属沭阳医院,江苏沭阳223600

出　　处：《陕西医学杂志》2024年第12期1604-1610,共7页Shaanxi Medical Journal

基　　金：国家自然科学基金资助项目(81860218);国家中医药管理局“胃癌毒邪论治”重点研究室开放课题(202135)。

摘　　要：目的:探讨长链非编码RNA(lncRNA) F-box富含亮氨酸的重复蛋白19反义RNA 1(FBXL19-AS1)调节微小RNA(miR)-342-3p/自噬相关蛋白10(ATG10)轴对结直肠癌(CRC)细胞增殖、迁移和奥沙利铂耐药性的影响。方法:实时定量聚合酶链反应(RT-qPCR)检测组织及正常肠上皮细胞(NCM460)、人CRC细胞系RKO、HCT-8细胞及HCT-8/L-OHP细胞中FBXL19-AS1、miR-342-3p、ATG10 mRNA表达水平。将HCT-8细胞分为shFBXL19-AS1组、shRNA组、shFBXL19-AS1+inhibitor NC组和shFBXL19-AS1+miR-342-3p inhibitor组,并以未转染的细胞为空白组。将HCT-8/L-OHP细胞分为L-OHP+shFBXL19-AS1组、L-OHP+shRNA组、L-OHP+shFBXL19-AS1+inhibitor NC组和L-OHP+shFBXL19-AS1+miR-342-3p inhibitor组,并以未转染的细胞为空白组。细胞计数试剂盒8(CCK-8)检测HCT-8、HCT-8/L-OHP细胞增殖及其耐药性。划痕实验检测细胞迁移能力。免疫印迹法(Western blot)检测P-糖蛋白(P-gp)、增殖蛋白(Ki-67)、迁移侵袭增强子因子1(MIEN1)、ATG10蛋白表达水平。双荧光素酶报告基因实验分析miR-342-3p与FBXL19-AS1、ATG10靶向关系。结果:FBXL19-AS1、ATG10 mRNA表达在HCT-8/L-OHP细胞及CRC组织中增加,miR-342-3p表达则降低(均P<0.05)。shFBXL19-AS1组细胞增殖、细胞迁移、FBXL19-AS1和ATG10 mRNA表达以及Ki-67、MIEN1、ATG10蛋白表达较shRNA组和空白组降低,miR-342-3p表达则增加(均P<0.05)。shFBXL19-AS1+miR-342-3p inhibitor组细胞增殖、细胞迁移、ATG10 mRNA以及Ki-67、MIEN1、ATG10蛋白表达较shFBXL19-AS1+inhibitor NC组增加,miR-342-3p表达则降低(均P<0.05)。L-OHP+shFBXL19-AS1组细胞增殖、细胞迁移、细胞耐药性及P-gp、Ki-67、MIEN1、ATG10蛋白表达较L-OHP+shRNA组和空白组降低(均P<0.05)。L-OHP+shFBXL19-AS1+miR-342-3p inhibitor组细胞增殖、细胞迁移、细胞耐药性及P-gp、Ki-67、MIEN1、ATG10蛋白表达较L-OHP+shFBXL19-AS1+inhibitor NC组增加(均P<0.05)。结论:干扰lncRNA FBXL19-AS1通过上调miR-342-3p/ATG10轴抑制CRC细胞的增殖�Objective:To investigate the effects of long non-coding RNA(lncRNA)F-box and leucine rich repeat protein 19 antisense RNA 1(FBXL19-AS1)on proliferation,migration,and oxaliplatin resistance in colorectal cancer(CRC)cells by regulating the microRNA(miR)-342-3p/autophagy-related genes10(ATG10)axis.Methods:RT-qPCR was used to detect the expression levels of FBXL19-AS1,miR-342-3p and ATG10 mRNA in tissues and normal intestinal epithelial cells(NCM460),human CRC cell line RKO,HCT-8 cells,and HCT-8/L-OHP cells.HCT-8 cells were separated into shFBXL19-AS1 group,shRNA group,shFBXL19-AS1+inhibitor NC group,and shFBXL19-AS1+miR-342-3p inhibitor group,with non-transfected HCT-8 cells as the blank group.HCT-8/L-OHP cells were divided into L-OHP+shFBXL19-AS1 group,L-OHP+shRNA group,L-OHP+shFBXL19-AS1+inhibitor NC group,and L-OHP+shFBXL19-AS1+miR-342-3p inhibitor group,with the non-transfected group as the blank group.CCK-8 method was applied to detect the proliferation and drug resistance of HCT-8 cells and HCT-8/L-OHP cells.Scratch experiment was applied to detect cell migration ability.Western blot was applied to detect the expression levels of P-glycoprotein(P-gp),proliferative protein(Ki-67),migration and invasion enhancer factor 1(MIEN1)and ATG10 protein.Luciferase reporter gene was applied to analyze the targeting relationship between miR-342-3p and FBXL19-AS1,ATG10,respectively.Results:The expression of FBXL19-AS1 and ATG10 mRNA was obviously increased in HCT-8/L-OHP cells and CRC tissues,while the expression of miR-342-3p was obviously reduced(all P<0.05).The proliferation,migration,FBXL19-AS1 mRNA expression,Ki-67,MIEN1,and ATG10 protein expression in the shFBXL19-AS1 group were obviously reduced compared to the shRNA group and blank group,while the expression of miR-342-3p was obviously increased(all P<0.05).The proliferation,migration,ATG10 mRNA,Ki-67,MIEN1,and ATG10 protein expression in the shFBXL19-AS1+miR-342-3p inhibitor group were obviously increased compared to the shFBXL19-AS1+inhibitor NC group,while the ex

关 键 词：结直肠癌 长链非编码RNA F-box富含亮氨酸的重复蛋白19反义RNA 1 微小RNA-342-3p 自噬相关蛋白10 细胞增殖 细胞迁移 奥沙利铂 

分 类 号：R-33[医药卫生]