hsa_circ_0006950调控miR-124-3p/EZH2轴促进胰腺癌细胞增殖与迁移的机制研究  

Mechanism of hsa_circ_0006950 Regulating miR-124-3p/EZH2 Axis to Promote the Proliferation and Migration of Pancreatic Cancer Cells

作　　者：王军堂[1] 齐普良 张金刚[1] 阿吉德 何婧 WANG Jun-tang;QI Pu-liang;ZHANG Jin-gang;A Ji-de;HE Jing(Department of Emergency Surgery,Qinghai Provincial People’s Hospital,Xining 810007,China;Department of General Surgery,Qinghai Provincial People’s Hospital,Xining 810007,China;the Second Department of Internal Medicine,Qinghai Provincial Armed Police Corps Hospital,Xining 810014,China)

机构地区：[1]青海省人民医院急诊外科,西宁810007 [2]青海省人民医院普外科,西宁810007 [3]武警青海省总队医院内二科,西宁810014

出　　处：《现代检验医学杂志》2022年第5期93-99,共7页Journal of Modern Laboratory Medicine

基　　金：青海省卫生计生系统科研项目(2017-wjzdx-23):青海省卫生健康系统指导性科研项目(2020-wjzdx-27)。

摘　　要：目的 检测环状核糖核酸(circular RNA,cicrRNA)hsa_circ_0006950在胰腺癌(pancreatic cancer,PC)中的表达,探讨其对胰腺癌细胞增殖、迁移的影响及其潜在分子作用机制。方法 利用实时荧光定量PCR(qRT-PCR)检测hsa_circ_0006950在胰腺癌组织及细胞中的相对表达水平;转染hsa_circ_0006950干扰载体构建低表达细胞系,通过CCK-8实验、克隆斑点形成实验和Transwell实验检测hsa_circ_0006950对胰腺癌细胞增殖、克隆形成及迁移的影响;利用生物信息学网站预测hsa_circ_0006950的miRNA分子靶标及其调控网络(hsa_circ_0006950-miR-124-3p-EZH2),双荧光素酶基因报告实验验证hsa_circ_0006950和miR-124-3p,miR-124-3p和EZH2的靶向结合关系;转染过表达/干扰miR-124-3p和过表达EZH2细胞系,探究miR-124-3p和EZH2及hsa_circ_0006950调控miR-124-3p/EZH2轴对胰腺癌细胞克隆形成及迁移的影响。结果 胰腺癌组织中hsa_circ_0006950表达明显高于癌旁正常组织(3.57±0.52 vs 1.01±0.03),差异有统计学意义(t=21.980,P<0.001)。胰腺癌细胞PANC-1(7.51±0.62),AsPC-1(5.26±0.45),Capan-2(3.69±0.38),SW1990(3.25±0.32)and BXPC-3(3.86±0.35)中hsa_circ_0006950相对表达明显高于正常胰腺导管上皮细胞HPDE6-C7(1.00±0.01)中的表达,差异有统计学意义(F=88.585,P<0.001)。与对照组相比,干扰hsa_circ_0006950表达组细胞增殖能力(0.79±0.17 vs 1.83±0.42),克隆形成率(51.42%±5.84%vs 78.76%±13.65%)和迁移数目(104.64±24.73 vs 218.21±31.57)明显降低,差异均有统计学意义(t=3.976,3.190,4.905,P=0.016,0.033,0.008)。mi R-124-3p是hsa_circ_0006950的下游靶基因,EZH2是miR-124-3p的直接靶标。hsa_circ_0006950靶向负调控miR-124-3p,miR-124-3p靶向负调控EZH2。与对照组相比,过表达miR-124-3p组PC细胞增殖(0.21±0.16 vs1.75±0.47),克隆形成率(47.85%±4.13%vs 81.54%±2.33%)和细胞迁移数目(118.74±24.65 vs 202.36±31.45)明显降低,差异均有统计学意义(t=5.378,14.317,3.390,均P<0.001);共转染过表达EZH2后,mobjective To detect the expression of cicrRNA hsa_circ_0006950 in pancreatic cancer(PC)and investigate the effect of cicrRNA hsa_circ_0006950 on the proliferation and migration of PC cells and its potential molecular mechanism.Methods QRT-PCR was used to detect the relative expression level of hsa_circ_0006950 in pancreatic cancer tissues and cells.Low expression cell lines were constructed by transfection of hsa_circ_0006950 interference vector,and the effects of hsa_circ_0006950 on PC cell proliferation,clonal formation and migration were detected by CCK-8,clone spot formation assay and Transwell assay.Predicted miRNA molecular target and regulatory network of hsa_circ_0006950(hsa_circ_0006950-miR-124-3p-EZH2)by bioinformatics website,targeted binding relationship between hsa_ circ_0006950 and miR-124-3p,miR-124-3p and EZH2 was verified by double luciferase gene report experiment.Over expression/interference of miR-124-3p and over expression of EZH2 cell lines were transfected to explore the effects of miR-124-3p,EZH2 and hsa_circ_0006950 regulating miR-124-3p/EZH2 axis on the formation and migration of PC cell clones.Results The expression of Hsa_circ_0006950 in PC tissues(3.57±0.52 vs 1.01±0.03)was significantly higher than that in adjacent normal tissues,the difference was statistically significant(t=21.980,P<0.001).In PC cell PANC-1(7.51±0.62),AsPC-1(5.26±0.45),Capan-2(3.69±0.38),SW1990(3.25±0.32),the relative expression of hsa_circ_0006950 in BXPC-3(3.86±0.35)was significantly higher than that in HPDE6-C7(1.00±0.01)in normal pancreatic duct epithelial cells,the difference was statistically significant(F=88.585,P<0.001).Compared with the control group,the proliferation ability and clone formation rate of hsa_circ_0006950 expression group were 0.79±0.17 vs 1.83±0.42,and 51.42±5.84 vs 78.76±13.65 and the number of migrations(104.64±24.73 vs 218.21±31.57)were significantly decreased,the differences were statistically significant(t =3.976,3.190,4.905,P =0.016,0.033,0.008).MiR-124-3p is the downstr

关 键 词：胰腺癌 hsa_circ_0006950 微小核糖核酸-124-3p EZH2 增殖 克隆形成 迁移 

分 类 号：R736.7[医药卫生—肿瘤] R730.43[医药卫生—临床医学]