circ-PKD2靶向miR-372-3p对结直肠癌细胞增殖、迁移和侵袭的影响  

Effect of circ-PKD2 on the proliferation,migration and invasion of colorectal cancer cells by targeting miR-372-3p

作　　者：何峰 罗少锋[1] 王威[1] 赵磊[2] 王译文 张兴 HE Feng;LUO Shao-feng;WANG Wei;ZHAO Lei;WANG Yi-wen;ZHANG Xing(Department of Clinical Laboratory,Puai Hospital,Tongji Medical College,Huazhong University of Science and Technology/Wuhan Fourth Hospital,Wuhan 430033,China;Department of Gastrointestinal Surgery,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China)

机构地区：[1]华中科技大学同济医学院附属普爱医院/武汉市第四医院检验科,武汉430033 [2]华中科技大学同济医学院附属协和医院胃肠外科,武汉430022

出　　处：《临床与实验病理学杂志》2022年第8期903-909,共7页Chinese Journal of Clinical and Experimental Pathology

基　　金：国家自然科学基金(81702392)。

摘　　要：目的 探讨环状RNA(circular RNA,circRNA)-PKD2在结直肠癌细胞增殖、迁移和侵袭中发挥的作用及可能的调控机制。方法 采用qRT-PCR分别检测结直肠癌组织、癌旁组织、结直肠癌细胞系(HCT116、HT-29、SW480、LoVo)与正常肠黏膜上皮细胞(HIEC)中circ-PKD2的表达水平。应用MTS实验、划痕愈合实验和Transwell实验分别检测各组细胞的增殖、迁移和侵袭能力。运用qRT-PCR和Western blot法分别检测circ-PKD2靶基因及下游蛋白的表达。结果 circ-PKD2在结直肠癌和癌旁组织中的表达量分别为1.28±0.14和6.38±0.29,circ-PKD2在结直肠癌组织中显著低表达(P<0.01)。与正常肠黏膜上皮细胞(HIEC)比较,circ-PKD2在结直肠癌细胞系中显著低表达(P<0.01),其表达水平最低的细胞系是HT-29细胞(P<0.01)。NC组和circ-PKD2组HT-29细胞中circ-PKD2的表达量分别为1.02±0.11和12.02±1.08,差异有统计学意义(P<0.01)。与NC组比较,circ-PKD2组HT-29细胞的增殖能力明显降低(P<0.05)。NC组和circ-PKD2组HT-29细胞划痕愈合率分别为(69.65±5.99)%和(25.58±4.33)%,NC组和circ-PKD2组HT-29细胞侵袭数分别为(89.28±11.07)个和(41.20±10.33)个。与NC组比较,circ-PKD2组HT-29细胞的迁移、侵袭能力明显降低(P均<0.01)。starBase v2.0软件分析circ-PKD2与miR-372-3p存在互补结合位点。circ-PKD2能够靶向结合miR-372-3p(P<0.01)。结直肠癌组织中circ-PKD2与miR-372-3p的表达呈明显负相关(P<0.01)。与NC组比较,circ-PKD2组HT-29细胞中miR-372-3p表达显著降低(P<0.01)。LATS2蛋白和Hippo信号通路蛋白的表达显著增加。结论 circ-PKD2在结直肠癌组织和细胞系中低表达,circ-PKD2通过靶向miR-372-3p抑制结直肠癌HT-29细胞的增殖、迁移和侵袭能力。Purpose To explore the role of circular RNA(circRNA)-PKD2 in the proliferation,migration and invasion of colorectal cancer cells and its possible regulatory mechanisms.Methods Real-time quantitative polymerase chain reaction(qRT-PCR) was used to detect the expression level of circ-PKD2 in colorectal cancer and adjacent tissues,colorectal cancer cell lines(HCT116,HT-29,SW480,LoVo) and normal intestinal mucosal epithelial cells(HIEC).The cell line with the lowest expression was selected as the research object,and the negative control plasmid(NC group) and circ-PKD2 overexpression plasmid(circ-PKD2 group) were transfected respectively.qRT-PCR was used to detect the expression level of circ-PKD2 in each group of cells.MTS experiment,scratch healing experiment and Transwell experiment were used to detect the proliferation,migration and invasion ability of each group of cells.Based on the bioinformatics software starBase v2.0,the target gene of circ-PKD2 was predicted.The dual luciferase reporter gene experiment was carried out to verify the targeting relationship.Pearson correlation analysis was performed on the expression of circ-PKD2 and target gene in colorectal cancer tissues.qRT-PCR and Western blot were used to detect the expression of circ-PKD2 target gene and downstream proteins.Results The expression of circ-PKD2 in colorectal cancer and para-cancerous tissues was 1.28±0.14 and 6.38±0.29,respectively.circ-PKD2 was significantly lower expressed in colorectal cancer tissues(P<0.01).Compared with normal intestinal mucosal epithelial cells(HIEC),circ-PKD2 was significantly lower expressed in colorectal cancer cell lines(P<0.01),and the cell line with the lowest expression level was HT-29 cell(P<0.01).The expression of circ-PKD2 in HT-29 cells in the NC group and circ-PKD2 group were 1.02±0.11 and 12.02±1.08,respectively,and the difference was statistically significant(P<0.01).Compared with the NC group,the proliferation ability of HT-29 cells in the circ-PKD2 group was significantly reduced(P<0.05).The wound

关 键 词：结直肠肿瘤 circ-PKD2 miR-372-3p 增殖 迁移 侵袭 

分 类 号：R735.3[医药卫生—肿瘤]