miR-505-3p对胃癌细胞增殖、克隆形成及迁移、侵袭的影响及机制研究  

Effect and Mechanism of miR-505-3p on Proliferation,Clone Formation,Migration and Invasion of Gastric Cancer Cells

作　　者：张金刚[1] 齐普良 吴刚 郭厚乐 王磊[4] ZHANG Jin-gang;QI Pu-liang;WU Gang;GUO Hou-le;WANG Lei(Department of Emergency Surgery,Qinghai Provincial People’s Hospital,Xining 810000,China;Department of General Surgery,Qinghai Provincial People’s Hospital,Xining 810000,China;Department of Surgical Oncology,Qinghai Provincial People’s Hospital,Xining 810000,China;Department of Pathology,Qinghai Provincial People’s Hospital,Xining 810000,China)

机构地区：[1]青海省人民医院急诊外科,西宁810000 [2]青海省人民医院普外科,西宁810000 [3]青海省人民医院肿瘤外科,西宁810000 [4]青海省人民医院病理科,西宁810000

出　　处：《现代检验医学杂志》2022年第4期69-74,153,共7页Journal of Modern Laboratory Medicine

基　　金：青海省2018年医药卫生科技项目课题(指导性项目)(No.Y2018-21)。

摘　　要：目的检测胃癌组织及细胞中miR-505-3p表达,并探究其对胃癌细胞增殖、克隆形成、迁移和侵袭的影响及潜在分子机制。方法利用实时荧光定量PCR实验(qRT-PCR)检测胃癌组织、癌旁正常组织及胃癌细胞和人正常胃黏膜细胞中miR-505-3p相对表达;构建miR-505-3p过表达、c-MYC过表达和敲低表达细胞系,通过CCK-8法,克隆斑点形成实验、Transwell侵袭/迁移实验检测miR-505-3p和c-MYC对胃癌细胞增殖、克隆形成及迁移、侵袭的影响;裸鼠皮下成瘤实验验证miR-505-3p对裸鼠肿瘤生长的影响;双荧光素报告实验验证miR-505-3p和c-MYC的靶向关系,并探究其两者间的表达调控作用;Western blot检测miR-505-3p和c-MYC对Wnt/β-catenin通路关键蛋白表达的影响。结果临床胃癌组织中miR-505-3p表达水平较正常癌旁组织显著下调(0.92±0.37 vs 1.74±0.59),差异有统计学意义(t=3.723,P<0.01)。胃癌细胞系MGC803(1.12±0.35),AZ521(2.31±0.24)和HGC-27(2.28±0.43)中miR-505-3p相对表达较人正常胃黏膜细胞系GES1(4.62±0.79)明显降低,差异有统计学意义(F=26.109,P<0.001)。miR-505-3p过表达组细胞增殖(0.92±0.27)、克隆形成(51.67±21.75)、迁移(43.25±15.47)、侵袭(38.53±14.76)能力较NC组(1.85±0.34,128.36±36.42,100.08±2.12,100.12±1.94)明显降低,差异均有统计学意义(t=3.131~7.166,均P<0.05);miR-505-3p过表达抑制了裸鼠生长。c-MYC是miR-505-3p的靶基因,miR-505-3p靶向负调控c-MYC。c-MYC过表达组细胞增殖(2.72±0.68)、迁移(147.15±20.36)、侵袭(145.46±22.73)能力较NC组(1.85±0.34,100.08±2.12,100.12±1.94)明显增高,差异均有统计学意义(t=2.455~4.456,均P<0.05);c-MYC过表达可逆转miR-505-3p对胃癌细胞增殖、迁移和侵袭的抑制作用。miR-505-3p过表达组Wnt(0.46±0.07),β-catenin(0.50±0.05)蛋白相对表达水平明显低于NC组(1.01±0.02,1.02±0.02),差异均有统计学意义(t=8.139,7.342,均P<0.001);过表达c-MYC能够逆转miR-505-3p对Wnt和β-catenin蛋�Objective The expression of miR-505-3p in gastric cancer tissues and cells was detected,and its effect on proliferation,clonogenesis,migration and invasion of gastric cancer cells and its potential molecular mechanism were explored.Methods QRT-PCR was used to detect the relative expression of miR-505-3p in gastric cancer tissues,normal adjacent tissues,gastric cancer cells and human normal gastric mucosa cells.MiR-505-3p overexpression,c-MYC overexpression and knockdown expression cell lines were constructed,and the effects of miR-505-3p and c-MYC on proliferation,clonal formation,migration and invasion of gastric cancer cells were detected by CCK-8 assay,clone spot formation assay and Transwell invasion/migration assay.The effect of miR-505-3p on tumor growth in nude mice was detected by subcutaneous tumorigenesis assay.Dual fluorescein reporting assay was used to verify the targeting relationship between miR-505-3p and c-MYC,and explored the expression regulation between the two.Western blot was used to detect the effects of miR-505-3p and c-MYC on the expression of key proteins in the Wnt/β-catenin pathway.Results The expression level of miR-505-3p in gastric cancer tissues was significantly down-regulated compared with normal adjacent tissues(0.92±0.37 vs 1.74±0.59),the difference was statistically significant(t=3.723,P<0.01).The relative expression of miR-505-3p in gastric cancer cell line MGC803(1.12±0.35),AZ521(2.31±0.24),and HGC-27(2.28±0.43)was significantly lower than that in human normal gastric mucosa cell line GES1(4.62±0.79),the differences was statistically significant(F=26.109,P<0.001).MiR-505-3p overexpression group showed proliferation(0.92±0.27),clonal formation(51.67±21.75),migration(43.25±15.47)and invasion ability(38.53±14.76)were significantly decreased compared with NC group(1.85±0.34,128.36±36.42,100.08±2.12,100.12±1.94),the differences were statistically significant(t=3.131~7.166,all P<0.05).MiR-505-3p overexpression inhibited the growth of nude mice.C-MYC was the target g

关 键 词：胃癌 微小核糖核酸-505-3p C-MYC 增殖 克隆形成 迁移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤] R730.4[医药卫生—临床医学]