miR-513a-3p在结肠癌中高表达并诱导细胞生长和迁移的机制研究  

Studies on the mechanism of miR-513a-3p overexpression and induction of cell growth and migration in colorectal cancer

作　　者：缪作华[1] 刘素云 李丹 Miao Zuohua;Liu Suyun;Li Dan(Dept of Pathology, Golden Hospital, The First Affiliated Hospital of Gannan Medical College, Ganzhou 341000;The First Affiliated Hospital of Nanchang University, Nanchang 330000)

机构地区：[1]赣南医学院第一附属医院黄金院区病理科,赣州341000 [2]南昌大学第一附属医院,南昌330000

出　　处：《安徽医科大学学报》2021年第7期1088-1093,共6页Acta Universitatis Medicinalis Anhui

基　　金：江西省卫生计生委科技计划项目(编号:20195362)。

摘　　要：目的观察miR-513a-3p在结肠癌组织与癌旁正常组织的表达差异,探讨miR-513a-3p诱导结肠癌(CRC)细胞生长和迁移的机制。方法收集CRC患者30例作为研究对象,原位杂交和实时荧光定量PCR(qRT-PCR)分别检测CRC组织与癌旁组织miR-513a-3p的表达水平。取人CRC细胞株HT-29和SW480培养后,转染miR-513a-3p类似物作为促进表达组,设置阴性对照组,四甲基偶氮唑盐(MTT)比色法实验两组细胞活力,Transwell法检测细胞侵袭情况,细胞划痕实验检验细胞迁移能力,Western blot法检测NF-κB和Wnt/β-catenin通路相关蛋白表达水平。结果原位杂交检测结果显示,CRC组织miR-513a-3p阳性表达高于癌旁组织。qRT-PCR检测结果显示,CRC组织和癌旁组织miR-513a-3p相对表达量分别为(0.37±0.09)和(1.01±0.16),差异有统计学意义(P<0.01)。与阴性对照组比较,促进表达组细胞活力在3、4 d中均升高(P<0.01),促进表达组的HT-29和SW480细胞侵袭细胞比例升高(P<0.01),促进表达组HT-29和SW480的细胞迁移能力增强。Western blot结果显示,与阴性对照组比较,促进表达组的HT-29细胞和SW480细胞p-p65、p-IκBα、Wnt3a、Wnt5a和β-catenin蛋白相对表达量均升高(P<0.01)。结论CRC组织miR-513a-3p表达异常升高,过表达的miR-513a-3p促进CRC细胞生长、迁移和侵袭作用,其作用机制可能与NF-κB通路和Wnt/β-catenin通路激活有关。Objective To investigate the expression of miR-513a-3p in colorectal cancer tissues and adjacent normal tissues,and to explore the mechanism of miR-513a-3p inducing colorectal cancer(CRC)cell growth and migration.Methods The total of 30 patients with colon cancer were selected as the research objects.The expression levels of miR-513a-3p in colon cancer tissues and adjacent tissues were detected by in situ hybridization and real-time fluorescence quantitative PCR.CRC cell lines HT-29 and SW480 were cultured and transfected with miR-513a-3p analogues as the expression promoting group.The negative control group was set up.The cell viability was detected by tetramethyl azo salt(MTT)colorimetry,the cell invasion was detected by Transwell method and cell migration was detected by cell scratch test.The expression of NF-κB and Wnt/β-Catenin pathway related proteins was detected by Western blot.Results The results of situ hybridization showed that the positive expression of miR-513a-3p in colon cancer tissues was higher than that in adjacent tissues.The results of qRT-PCR showed that the relative expression of mir-513a-3p was(0.37±0.09)in colon cancer tissues and(1.01±0.16)in adjacent tissues,and the difference was statistically significant(P<0.01).Compared with the negative control group,the cell viability of the promoted expression group increased in 3 and 4 d(P<0.01),the invasion rate of HT-29 and SW480 cells in the promoting expression group increased(P<0.01),and the cell migration ability of HT-29 and SW480 in the promoting expression group was enhanced.The results of Western blot showed that compared with the negative control group,the relative expressions of p-p65,p-IκBα,Wnt3a,Wnt5a andβ-Catenin protein in HT-29 cells and SW480 cells significantly increased(P<0.01).Conclusion The overexpression of miR-513a-3p promotes the growth,migration and invasion of colon cancer cells.The possible mechanism is related to the activation of NF-κB pathway and Wnt/β-Catenin pathway.

关 键 词：miR-513a-3p 结肠癌 细胞迁移 NF-κB和Wnt/β-catenin信号通路 

分 类 号：R735.35[医药卫生—肿瘤] R331.1[医药卫生—临床医学]