miR-411-5p在胃癌中的表达及其生物学功能

Expression and biological function of miR-411-5p in gastric cancer

目的检测miR-411-5p(hsa-microRNA-441-5p)在胃癌组织及胃癌细胞中表达情况以及对胃癌细胞功能的影响。方法采用q-PCR检测48对胃癌及癌旁组织、人胃癌细胞系BGC-823、BGC-803、SGC-7901、AGS以及人胃粘膜上皮细胞株GES-1中miR-411-5p的表达水平;采用miR-411-5p expression mimic或inhibitor转染表达差异最显著细胞后,通过平板克隆、CCK-8法、流式细胞仪检测miR-411-5p对胃癌细胞的增殖、迁移、凋亡的影响。结果胃癌组织中miR-411-5p的相对表达量(0.50±0.25)明显低于癌旁组织(1.19±0.44,t=-9.55,P<0.01);与胃正常上皮细胞GES-1(0.90±0.15)中的miR-411-5p相比,人胃癌细胞系BGC-823(0.39±0.08,t=9.33,P<0.01)、BGC-803(0.55±0.15,t=5.21,P<0.01)、SGC-7901(0.58±0.07,t=6.03,P<0.01)、AGS(0.66±0.10,t=4.02,P<0.01)表达相对较低; BGC-823细胞过表达miR-411-5p后,细胞中miR-411-5p的表达(4.00±0.59)较阴性处理组(0.86±0.06)或空白对照组(0.95±0.27)明显增高(t=-15.90、-15.52,P<0.01,转染组(119.60±19.23)克隆形成能力较阴性对照组(222.40±24.82)及空白对照组(242.60±17.39)明显减弱(F=50.67,P<001),CCK-8实验亦显示其增殖能力明显减弱,细胞凋亡实验显示转染组凋亡率(8.72±0.66)%较阴性对照组(1.21±0.18)%及空白对照组(1.10±0.27)%略微上升(P<0.01);转染组跨膜细胞数(348.80±24.13)较阴性对照组(908.40±57.94)和空白对照组显著减少(952.20±51.30,F=250.07,P<0.01)。结论在胃癌组织和细胞中,miR-411-5p是一个抑制性miRNA,其表达水平下调参与了胃癌的发生和发展,有可能成为胃癌生物靶向治疗的新靶点。

Objective To detect the expression of miR-411-5p(hsa-microRNA-441-5p)in gastric cancer tissues and gastric cancer cells,and investigate its effect on functions of gastric cancer cells.Methods The expression levels of miR-411-5p in gastric cancer,adjacent tissues,gastric cancer cell lines of human(BGC-823,BGC-803,SGC-7901,AGS)and human gastric mucosa epithelial cell lines GES-1 were detected by q-PCR.The effects of miR-411-5p on proliferation,migration and apoptosis of gastric cancer cells were detected by flatscreen cloning,CCK-8 and flow cytometry after the cells were transfected with mir-411-5p expression mimic or inhibitor.Results The relative expression of mir-411-5p in gastric cancer tissues(0.50±0.25)was significantly lower than that in adjacent tissues(1.19±0.44,t=-9.55,P<0.01).Compared with mir-411-5p in normal gastric epithelial GES-1(0.90±0.15),human gastric cancer cell line BGC-823(0.39±0.08,t=9.33,P<0.01),BGC-803(0.55±0.15,t=5.21,P<0.01),SGC-7901(0.58±0.07,t=6.03,P<0.01),AGS(0.66±0.10,t=4.02,P<0.01)showed relatively lower expression.After overexpression of miR-411-5p in BGC-823 cells,the expression of miR-411-5p in cells(4.00±0.59)was higher than that in the negative treatment group(0.86±0.06)and the blank control group(0.95±0.27).In the transfection group(119.60±19.23),the clonal formation ability was significantly reduced compared with the negative control group(222.40±24.82)and the blank control group(242.60±17.4)(F=50.67,P<0.01)the CCK-8 experiment also showed a significant reduction in proliferation capacity,and the cell apoptosis experiment showed that the number of transmembrane cells(348.80±24.13)in the transfection group was significantly lower than that of the negative control group(908.40+57.94)and the blank control group(952.20±51.30,F=250.07,P<0.01).Conclusion In gastric cancer tissues and cells,mir-411-5p was an inhibitory miRNA,and its down-regulated expression level was involved in the occurrence and development of gastric cancer,which may become a new target for biological targeted treatment of gastric cancer.