Hsa-miR-654-5p调控结肠癌细胞恶性生物学行为及临床意义研究

Research on the regulation of malignant biological behavior of colon cancer cells by Hsa miR-654-5p and its clinical significance

目的检测结肠癌中Hsa-miR-654-5p表达,分析其对结肠癌细胞恶性生物学行为的影响及临床意义。方法采用qRT-PCR法检测结肠癌细胞(HT-29、Caco-2、SW480、HCT116、LOVO)与人正常结肠上皮细胞(HCoEpiC),2020年1月—2022年12月在洛阳市中心医院中心实验室收集的90例结肠癌与癌旁组织中Hsa-miR-654-5p表达,分析其与患者临床病理特征及预后的关系。按照简单随机分组法进行实验分组:NC组、miR-con组、Hsa-miR-654-5p过表达组及敲减组(每组n=10)。采用噻唑蓝比色法、克隆形成实验、细胞划痕试验和Transwell实验测定各组细胞增殖、克隆、迁移、侵袭变化,蛋白印迹法检测E-钙黏蛋白、人波形蛋白及神经性钙黏附蛋白表达。结果Hsa-miR-654-5p在结肠癌细胞及组织中表达量明显增高,且不同Hsa-miR-654-5p表达水平患者肿瘤大小、分化程度、TNM分期及淋巴结转移比较差异均有统计学意义(P<0.05),而年龄、性别、病理学类型及癌胚抗原比较差异均无统计学意义(P>0.05)。与NC组和miR-con组相比,Hsa-miR-654-5p过表达组SW480细胞增殖及克隆能力、细胞迁移率、穿膜细胞数、E-钙黏蛋白、人波形蛋白及神经性钙黏附蛋白表达均显著增高,而敲减组细胞上述指标明显降低,组间差异均有统计学意义(P<0.05)。Hsa-miR-654-5p高表达患者中位生存期显著低于低表达者(20.44个月vs.38.16个月,χ^(2)=10.079,P<0.001)。结论Hsa-miR-654-5p在结肠癌中高表达,且与患者临床病理特征及预后相关,敲减表达后可抑制结肠癌细胞恶性生物学行为,机制可能与抑制上皮-间充质转化有关。

Objective Detect the expression of Hsa-miR-654-5p in colon cancer,analyze its impact on the malignant biological behavior of colon cancer cells and its clinical significance.Methods Detect the expression of Hsa-miR-654-5p in colon cancer cells(HT-29,Caco-2,SW480,HCT116,LOVO)and human normal colon epithelial cell(HCoEpiC),as well as in 90 cases of colon cancer and adjacent tissues collected from the central laboratory of Luoyang Central Hospital from January 2020 to December 2022 by qRT-PCR,analyze the relationship between its expression and clinical pathological characteristics and prognosis of patients.Conduct experimental grouping according to the simple random grouping method:NC group,miR-con group,Hsa-miR-654-5p overexpression group and knockdown group(n=10 per group).Cell proliferation,cloning,migration,and invasion changes in each group were measured by thiazole blue colorimetry,clone formation assay,cell scratch assay,and Transwell assay,detected the expression of Epithelia cadherin,human vimentin,and Nervous cadheri by Western blotting.Results The expression level of Hsa-miR-654-5p was significantly increased in colon cancer cells and tissues.There were significant differences in tumor size,differentiation,TNM staging,and lymph node metastasis(P<0.05),while there were no statistically significant differences in age,gender,pathological type,and carcinoembryonic antigen(P>0.05).The proliferation and clone ability,cell migration rate,number of transmembrane cells,and protein expression of Epithelia cadherin,human vimentin,and Nervous cadherin of SW480 cells in the Hsa-miR-654-5p overexpression group were significantly increased when compared with the NC group and miR-con group,while the knockdown group showed a significant decrease in the above indicators,with significant differences between groups(P<0.05).The median survival of colon cancer patients with high expression of Hsa-miR-654-5p was significantly lower than those with low expression(20.44 months vs.38.16 months,χ^(2)=10.079,P<0.001).Conclusion Hsa-miR-654-5p is highly expressed in colon cancer and significantly correlated with clinicopathological features and prognosis of patients.Knockdown of it can inhibit the malignant biological behavior of colon cancer cells,which may be related to the inhibition of epithelial-mesenchymal transition.