MiR-181c-5p对卵巢癌干细胞样细胞肿瘤血管生成拟态的作用及其机制  

Effect of miR-181c-5p on vasculogenic mimicry in ovarian cancer stem-like cells and its mechanism

作　　者：吴颖颖 文小玲 夏玉芳[1] 于啸[1] 娄艳辉[1] WU Yingying;WEN Xiaoling;XIA Yufang;YU Xiao;LOU Yanhui(Department of Gynecology,The Affiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学附属医院妇科,山东青岛266003 [2]佛山市第一人民医院妇产科

出　　处：《精准医学杂志》2024年第3期247-251,256,共6页Journal of Precision Medicine

基　　金：青岛市市南区科技计划项目(2022-2-006-YY)。

摘　　要：目的探讨miR-181c-5p对卵巢癌干细胞样细胞(OCS-LCs)肿瘤血管生成拟态(VM)的作用及其机制。方法采用无血清悬浮培养法将人卵巢癌细胞系OVCAR3细胞诱导形成OCS-LCs。将OVCAR3细胞分为A~C组,各组分别转染NC-miR-181c-5p、siRNA-miR-181c-5p和pRNA-miR-181c-5p。通过成球实验评估A~C组细胞成球能力。采用实时荧光定量PCR(RT-qPCR)方法检测A~C组细胞miR-181c-5p的相对表达量,采用Western blot实验检测A~C组细胞Oct-4、Nanog、HIF-1α和VEGF蛋白相对表达量。采用CCK-8实验检测A~C组细胞的活性,采用三维立体培养实验检测A~C组的血管形成率。结果OVCAR3细胞成功被诱导形成OCS-LCs。RT-qPCR实验结果显示,B组细胞的miR-181c-5p相对表达量显著低于A组,C组高于A组(t=2.25、8.68,P<0.05)。成球实验结果显示,B组与A组、A组与C组相比,细胞的成球周期显著缩短,最大的细胞球直径显著增大,成球率显著增加(t=5.56~33.66,P<0.05)。Western blot实验结果表明,B组与A组、A组与C组相比,Oct-4、Nanog、HIF-1α和VEGF蛋白相对表达量显著升高(t=4.51~56.15,P<0.05)。CCK-8实验结果显示,B组的细胞活性高于A组,C组低于A组(F=97.70~281.80,P<0.05)。三维立体培养实验结果显示,B组与A组、A组与C组相比较,血管形成率显著性提高(t=3.70、18.67,P<0.05)。结论miR-181c-5p可能通过降低细胞中HIF-1α和VEGF蛋白的表达,从而抑制OCS-LCs的VM形成。Objective To investigate the effect of miR-181c-5p on vasculogenic mimicry(VM)in ovarian cancer stem-like cells(OCS-LCs)and its mechanism.Methods OVCAR3 cells were induced into OCS-LCs by serum-free suspension culture.OVCAR3 cells were divided into groups A,B,and C and were transfected with NC-miR-181c-5p,siRNA-miR-181c-5p,and pRNA-miR-181c-5p,respectively.The sphere-forming experiment was used to assess the sphere formation ability of groups A,B,and C.RT-qPCR was used to measure the relative expression level of miR-181c-5p in cells of groups A,B,and C,and Western blot was used to measure the relative protein expression levels of Oct-4,Nanog,HIF-1α,and VEGF in cells of groups A,B,and C.CCK-8 assay was used to measure the viability of cells in groups A,B,and C,and the three-dimensional culture experiment was used to measure the angiogenesis rate of groups A,B,and C.Results OVCAR3 cells were successfully induced into OCS-LCs.RT-qPCR showed that group B had a significantly lower relative expression level of miR-181c-5p than group A,and group C had a significantly higher relative expression level than group A(t=2.25,8.68,P<0.05).The sphere-forming experiment showed that compared with group A,group B had significant decreases in sphere-forming cycle,and increases in maximum sphere diameter,and sphere-forming rate,and compared group A with group C,these indicators had the same changes(t=5.56-33.66,P<0.05).Western blot showed that compared with group A,group B had significant increases in the relative protein expression levels of Oct-4,Nanog,HIF-1α,and VEGF,and compared with group C,group A had significant increases in the relative expression levels of these proteins(t=4.51-56.15,P<0.05).CCK-8 assay showed that group B had a significantly higher cell viability than group A,and group C had a significantly lower cell viability than group A(F=97.70-281.80,P<0.05).The three-dimensional culture experiment showed that group B had a significant increase in angiogenesis rate compared with group A,and compared with group C,gro

关 键 词：卵巢肿瘤 肿瘤干细胞 微RNAs 缺氧诱导因子1 Α亚基 血管内皮生长因子类 新生血管化 病理性 体外培养技术 

分 类 号：R737.31[医药卫生—肿瘤] R364.3[医药卫生—临床医学]