miR-224-5p靶向GABRE对人结肠癌细胞系SW480增殖的影响

Effect of miR-224-5p targeting GABRE on proliferation of colorectal cancer cell line SW480

目的 探讨miR-224-5p通过靶点GABRE对结直肠癌细胞系增殖的抑制作用和可能的调控机制。方法 从基因表达综合数据库(GEO)中检索并筛选GPL18058的microRNA(miRNA)表达阵列,筛选出结直肠癌与正常组织差异表达的miR-224-5p后,通过Target Scan Human数据库预测其下游靶点GABRE,并利用UALCAN分析GABRE在结直肠癌组织中的表达情况和结直肠癌预后关系。利用实时定量PCR(qRT-PCR)检测结直肠癌细胞系中的miR-224-5p和GABRE的表达水平。在结直肠癌细胞系SW480中,沉默miR-224-5p后,CCK-8法与克隆形成实验分析其增殖情况。RT-PCR和Western blot检测miR-224-5p对GABRE表达的影响。在SW480细胞中低表达GABRE,评估其对结直肠癌调节中的miR-224-5p的选择性作用。结果 在结直肠癌组织和细胞系中,miR-224-5p的表达异常上调,抑制其表达后,SW480细胞的增殖能力降低(P<0.05)。在结直肠癌组织和细胞系的基因和蛋白质水平上,miR-224-5p正调控GABRE(P<0.05)。GABRE低表达抑制SW480细胞增殖(P<0.05)。结论 miR-224-5p促进结直肠癌中SW480细胞系的增殖,并通过正调控GABRE发挥作用。

Objective To investigate the inhibitory effect of miR-224-5p on the proliferation of colorectal cancer cell line and its possible regulatory mechanism by targeting GABRE. Methods microRNA(miRNA) expression arrays of GPL18058 were searched and screened from gene expression database(GEO). After screening out miR-224-5p differentially expressed between colorectal cancer and normal tissues, the downstream target GABRE was predicted by target scan human database, and the relationship between GABRE expression in colorectal cancer tissues was analyzed by using UALCAN. Quantitative real-time PCR was used to detect the expression of miR-224-5p and GABRE in colorectal cancer cell lines. In SW480, miR-224-5p was overexpressed. After silencing miR-224-5p in colorectal cancer cell line SW480,the proliferation of SW480 cells was analyzed by CCK-8 method and clone formation test. The effect of miR-224-5p on the expression of GABRE was detected by real-time PCR and Western blot. Then, GABRE was lower expressed in SW480 cells to evaluate its selective effect on miR-224-5p in colorectal cancer regulation. Results In colorectal cancer tissues and cell lines, the expression of miR-224-5p was upregulated, and inhibition of miR-224-5P reduced the proliferation ability of SW480 cells(P<0.05). miR-224-5p positively regulated GABRE at gene and protein levels in colorectal cancer tissues and cell lines(P<0.05). Low expression of GABRE inhibited the proliferation of SW480 cells(P<0.05). Conclusion miR-224-5p promote the proliferation of SW480 cell line, and plays a role by positive regulating GABRE.