摘要
目的探究CircPIP5K1A靶向调节miR-101-3p对结直肠癌(CRC)SW480细胞生长、迁移、侵袭和凋亡的影响。方法qRT-PCR检测人CRC组织、邻近正常组织、CRC细胞株SW480、正常结肠上皮细胞FHC中CircPIP5K1A、miR-101-3p水平;将si-NC、si-CircPIP5K1A、si-CircPIP5K1A+inhibitor NC、si-CircPIP5K1A+miR-101-3p inhibitor分别转染至SW480细胞并命名为si-NC组、si-CircPIP5K1A组、si-CircPIP5K1A+inhibitor NC组、si-CircPIP5K1A+miR-101-3p inhibitor组,未做任何处理的SW480细胞记为NC组。双荧光素酶报告基因实验验证CircPIP5K1A与miR-101-3p的关系;qRT-PCR检测SW480细胞中CircPIP5K1A、miR-101-3p表达;CCK-8法检测SW480细胞增殖情况;流式细胞术检测SW480细胞凋亡率;Transwell检测SW480细胞侵袭、迁移数量;Western blot检测SW480细胞E-cadherin、Vimentin、N-cadherin蛋白水平;小鼠异种移植检测肿瘤生长。结果与邻近正常组织[(1.00±0.03)、(1.00±0.02)]或FHC细胞(1.00±0.00)相比,CRC组织和SW480细胞CircPIP5K1A水平[(1.85±0.21)、(1.54±0.16)]显著上调(P<0.05),miR-101-3p表达[(0.31±0.02)、(0.36±0.04)]显著下调(P<0.05)。与NC组、si-NC组相比,si-CircPIP5K1A组SW480细胞A 450值、迁移、侵袭细胞数量、CircPIP5K1A表达、N-cadherin、Vimentin蛋白水平显著下降(P<0.05),SW480细胞凋亡率、miR-101-3p表达、E-cadherin蛋白水平显著升高(P<0.05);而抑制miR-101-3p表达减弱了沉默CircPIP5K1A抑制SW480细胞增殖、迁移和侵袭的效果;CircPIP5K1A负向调控miR-101-3p。与NC组、慢病毒空载(LV-NC)组相比,PIP5K1A慢病毒载体(LV-PIP5K1A)组肿瘤体积、肿瘤质量显著下降(P<0.05);与LV-PIP5K1A组、LV-PIP5K1A+拮抗剂对照(antiagomir)组相比较,LV-PIP5K1A+miR-101-3p拮抗剂(miR-101-3p antiagomir)组肿瘤体积、肿瘤质量显著升高(P<0.05)。结论沉默CircPIP5K1A可能通过上调miR-101-3p表达来对CRC细胞SW480增殖、迁移和侵袭产生影响。
Objective To investigate the effects of CircPIP5K1A targeting miR-101-3p on the growth,migration,invasion and apoptosis of colorectal cancer(CRC)SW480 cells.Methods CircPIP5K1A and miR-101-3p levels in human CRC tissues,adjacent normal tissues,CRC cell line SW480 and normal colon epithelial cells FHC were detected by qRT-PCR.si NC,si CircPIP5K1A,si CircPIP5K1A+inhibitor NC and si CircPIP5K1A+miR-101-3p inhibitor were respectively transfected into SW480 cells and named si NC group,si CircPIP5K1A group,si CircPIP5K1A+inhibitor NC group,si CircPIP5K1A+miR-101-3p inhibitor group,and SW480 cells without any treatment were labeled as NC group.Dual luciferase reporter gene assay verified the relationship between CircPIP5K1A and miR-101-3p;The expression of CircPIP5K1A and miR-101-3p in SW480 cells was detected by qRT-PCR;The proliferation of SW480 cells was detected by CCK 8 method;The apoptosis rate of SW480 cells was detected by flow cytometry;The invasion and migration of SW480 cells were detected by Transwell;The protein levels of E-cadherin,Vimentin and N-cadherin in SW480 cells were detected by Western blot;Tumor growth was detected by xenotransplantation in mice.Results Compared with the adjacent normal tissues[(1.00±0.03),(1.00±0.02)]or FHC cells(1.00±0.00),CircPIP5K1A levels in CRC tissues and SW480 cells[(1.85±0.21),(1.54±0.16)]were significantly up-regulated(P<0.05).The expression of miR-101-3p[(0.31±0.02),(0.36±0.04)]was significantly down-regulated(P<0.05).Compared with NC group and si NC group,the A450 value,number of migrating and invading cells,CircPIP5K1A expression,N-cadherin and Vimentin protein levels of SW480 cells in si CircPIP5K1A group were significantly decreased(P<0.05).The apoptosis rate,miR-101-3p expression and E-cadherin protein level of SW480 cells were significantly increased(P<0.05).Inhibition of miR-101-3p expression weakened the inhibitory effect of CircPIP5K1A on the proliferation,migration and invasion of SW480 cells.CircPIP5K1A negatively regulates miR-101-3p.Compared with NC
作者
刘建军
王攀
袁华燕
吴镇江
印隆宽
白翔宇
LIU Jianjun;WANG Pan;YUAN Huayan;WU Zhenjiang;YIN Longkuan;BAI Xiangyu(Department of Gastrointestinal Sugery,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan,China)
出处
《医学研究与战创伤救治》
北大核心
2023年第8期801-807,共7页
Journal of Medical Research & Combat Trauma Care
基金
川北医学院科研发展计划项目(7500520021)。
