MiR-1-3p通过抑制CAPRIN1调控胰腺癌发生发展

MiR-1-3p Regulates the Development of Pancreatic Cancer via Inhibiting CAPRIN1

目的:探讨miR-1-3p在胰腺癌发生发展中的分子机制。方法:以MIA-PaCa-2,SW 1990为研究目标,通过qRT-PCR技术检测miR-1-3p的表达量,利用TargetScan和miRDB数据库预测miR-1-3p的下游靶基因及结合位点,并通过构建双荧光素酶报告基因,进一步确认miR-1-3p与靶基因的结合。利用CCK8细胞增殖实验及平板克隆形成实验检测过表达miR-1-3p及敲低CAPRIN1对细胞增殖的作用;利用流式检测细胞周期;利用蛋白质免疫印迹方法检测miR-1-3p对CAPRIN1及其下游基因的影响;通过流式来确认,过表达miR-1-3p及敲减CAPRIN1基因对细胞周期的影响。结果:miR-1-3p在胰腺癌细胞MIA-PaCa-2,SW 1990中低表达;miR-1-3p直接与CAPRIN1的3'-untranslated region(3'-UTR)结合;过表达miR-1-3p或抑制CAPRIN1基因的表达可明显抑制胰腺癌细胞的增殖能力,同时也产生细胞周期阻滞。结论:miR-1-3p通过抑制CAPRIN1基因表达,而产生细胞周期阻滞进而抑制胰腺癌细胞的增殖能力。

Objective:To investigate the molecular mechanism ofmir-1-3p in the development and progression of pancreatic cancer.Methods:With MIA-PACA-2 and SW1990 as the research target,the expression level of mir-1-3p was detected by qRT-PCR technology,and the downstream target genes and binding sites of mir-1-3p were predicted by TargetScan and miRDB databases,and the double luciflucase reporter gene was constructed,the binding of miR-1-3p to target genes was further confirmed.CCK8 cell proliferation assay and plate clonal formation assay were used to detect the effects of overexpression of miR-1-3p and knockdown CAPRIN1 on cell proliferation.Western blot was used to detect the effect of miR-1-3p on CAPRIN1 and its downstream genes.Finally,the effects of miR-1-3p overexpression and CAPRIN1 knockdown on cell cycle were confirmed by flow cytometry.Results:In pancreatic cancer cells MIA-PACA-2,SW 1990,miR-1-3p was low in mRNA level.Mir-1-3p directly binds to the 3'-untranslated region(3'-UTR)of CAPRIN1.Overexpression of miR-1-3p or inhibition of CAPRIN1 gene expression can significantly inhibit the proliferation of pancreatic cancer cells,and also induce cell cycle arrest.Conclusion:MiR-1-3p inhibits the proliferation of pancreatic cancer cells by inhibiting the expression of CAPRIN1 gene,resulting in cell cycle arrest.