miR-590-5p靶向SMAD7促进结肠癌细胞进展的机制研究

Mechanism study on miR-590-5p in promoting progression of colon cancer cells by targeting SMAD7

目的研究miR-590-5p在结直肠癌组织和结肠癌细胞中的表达和功能及可能相关的分子机制。方法对2022年6月—2023年12月本院收治的进行手术的结直肠癌患者进行样本采集,共收集结直肠癌组织及癌旁组织临床样本40对,通过实时荧光定量聚合酶链反应(qPCR)检测miR-590-5p在两者的表达情况并与临床特征进行相关性分析。通过瞬时转染进行体外细胞实验,使用细胞计数试剂盒-8(CCK-8)检测miR-590-5p对结肠癌细胞增殖的作用,运用Transwell实验检测miR-590-5p对结肠癌细胞迁移与侵袭的影响。通过生物信息分析筛选数据库筛选miR-590-5p的靶基因SMAD7,分别采用双荧光素酶实验、qPCR和蛋白质免疫印迹实验(western blot)验证二者的作用及相关性。结果结直肠癌组织中miR-590-5p的表达水平与结直肠癌的临床分期、神经脉管侵犯情况和淋巴结转移数目情况存在正相关关系;过表达miR-590-5p会提高结肠癌细胞的增殖、迁移和侵袭(P<0.05),而抑制miR-590-5p表达后结果则相反(P<0.05);双荧光素酶实验结果表明SMAD7是miR-590-5p的一个靶基因,qPCR实验和western blot实验结果则表明二者存在负相关关系。结论miR-590-5p在进展期结直肠癌中可能扮演着积极的角色,其可能通过靶向SMAD7促进结肠癌细胞的增殖、迁移与侵袭。

Objective To investigate the expression and function of miR-590-5p in colorectal cancer tissues and colon cancer cells,along with the potential associated molecular mechanisms.Methods From June 2022 to December 2023,clinical samples were collected from patients with colorectal cancer who underwent surgery at our hospital.A total of 40 pairs of colorectal cancer tissue and adjacent tissue samples were collected for the detection of miR-590-5p expression by using real-time fluorescence quantitative polymerase chain reaction(qPCR),and its correlation with clinical features was analyzed.Transient transfection was used to carried out in vitro cellular experiments,and to study the impact of miR-590-5p on colon cancer cell proliferation using the Cell Counting Kit-8(CCK-8),as well as assessing its effects on cell migration and invasion using the Transwell assay.Through bioinformatic analysis screening,SMAD7 was identified as a potential target gene of miR-590-5p,and this association was verified using dual luciferase assay,qPCR,and western blot assays.Results The expression level of miR-590-5p in colorectal cancer tissues showed a positive correlation with clinical stage,neurovascular invasion,and the number of lymph node metastases in colorectal cancer.Over expression of miR-590-5p was found to enhance the proliferation,migration,and invasion of colon cancer cells(P<0.05),whereas inhibiting the expression of miR-590-5p had the opposite effect(P<0.05).Furthermore,the results of the dual luciferase assay indicated that SMAD7 was a target gene of miR-590-5p.Subsequent qPCR and western blot assays affirmed a negative correlation between them.Conclusions MiR-590-5p may play an active role in advanced colorectal cancer by targeting SMAD7 to promote the proliferation,migration and invasion of colon cancer cells.