微小RNA-27b-3p靶向干扰素调节因子4调控结肠癌细胞B细胞淋巴瘤-6表达的研究  

Modulation of B cell lymphoma/leukemia-6 by microRNA-27b-3p targeting interferon regulatory factor 4 in colon cancer cells

作　　者：赖斌[1] 胡林[1] 周小双 温华斌 李红浪[1] Lai Bin;Hu Lin;Zhou Xiaoshuang;Wen Huabin;Li Honglang(Department of Gastroenterology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,China;The Second Clinical College of Nanchang University,Nanchang 330029,China)

机构地区：[1]南昌大学第二附属医院肠胃外科,330006 [2]南昌大学第二临床学院,330029

出　　处：《中华实验外科杂志》2021年第1期164-167,共4页Chinese Journal of Experimental Surgery

基　　金：江西省教育厅科技项目(GJJ190073)。

摘　　要：目的观察微小RNA(microRNA,miR)-27b-3p与干扰素调节因子4(IRF4)的靶向关系,分析其对结肠癌细胞B细胞淋巴瘤分子-6(bcl-6)表达的影响。方法2015年12月至2017年1月在南昌大学第二附属医院确诊为结肠癌患者共53例,以肿瘤组织作为研究组,以正常结肠黏膜组织作为对照组。应用实时荧光定量聚合酶链反应(qPCR)检测miR-27b-3p的表达。应用免疫组织化学法检测结肠癌中干扰素调节因子4(IRF4)和B细胞淋巴瘤-6(bcl-6)的表达。选择人结肠癌细胞系116,构建阴性对照组、miR-27b-3p转染组、miR-27b-3p和IRF4共转染组,应用免疫印迹实验检测各组中IRF4和bcl-6的表达。应用双荧光素酶报告基因实验观察miR-27b-3p与IRF4的结合情况。组间比较采用t检验、配对t检验及方差分析。结果结肠癌中miR-27b-3p的表达(2.67±0.40)明显低于对照组(3.68±0.45,t=6.180,P<0.01),miR-27b-3p的表达在不同肿瘤最大径(≥5 cm为2.49±0.39,<5 cm为2.87±0.36)、浸润深度(浆膜及以外为2.44±0.34,未及浆膜为2.88±0.36)、脉管癌栓(有癌栓为2.28±0.42,无癌栓为2.77±0.31)、片状坏死(有坏死为2.36±0.42,无坏死为2.79±0.39)和淋巴结转移(有转移为2.37±0.41,无转移为2.81±0.22)分组的表达中差异有统计学意义(t=4.250、5.180、5.690、5.110、5.190,P值均<0.05)。miR-27b-3p与生存时间有关(χ^2=5.080,P<0.05),差异有统计学意义。miR-27b-3p与IRF4(r=-0.600,P<0.05)、miR-27b-3p与bcl-6(r=-0.690,P<0.05)均呈负相关性。双荧光素酶报告基因实验显示miR-27b-3p与IRF4具有靶向关系。miR-27b-3p转染组中IRF4和bcl-6的表达低于阴性对照组(t=3.240、4.330,P<0.05),差异有统计学意义,共转染组能逆转上述改变。结论miR-27b-3p靶向负调控IRF4调节结肠癌细胞bcl-6的表达,miR-27b-3p低表达是肿瘤形成和进展的重要事件,检测miR-27b-3p的表达可能与生存时间有关。Objective To observe the targeting relationship between microRNA(miRNA,miR)-27b-3p and interferon regulatory factor 4(IRF4),and analyze its effect on B cell lymphoma/leukemia-6(bcl-6)in colon cancer cells.Methods A total of 53 patients with colorectal carcinoma admitted at Second Affiliated Hospital of Nanchang University were selected from December 2015 to January 2017.Tumor tissues served as the study group,and normal colonic mucosa as the control group.The expression of miR-27b-3p was detected by real-time fluorescent quantitative polymerase chain reaction(PCR).The expression levels of IRF4 and bcl-6 were detected by immunohistochemistry.We selected human colorectal tumor116(HCT-116)cells of colon cancer,and constructed negative control group,miR-27b-3p transfection group,miR-27b-3p and IRF4 co-transfection group.IRF4 and bcl-6 were detected in different groups by Western blotting.The targeting relationship between miR-27b-3p and IRF4 was observed by double luciferase reporter gene assay.Results Expression of miR-27b-3p was significantly lower in colorectal cancer tissues(2.67±0.40)than that in the control group(3.68±0.45,t=6.180,P<0.01).Expression of miR-27b-3p was significantly different in tumor maximum diameter(≥5 cm:2.49±0.39,<5 cm:2.87±0.36),invasion depth(serosa and beyond:2.44±0.34,less serosa:2.88±0.36),vascular tumor thrombus(tumor thrombus:2.28±0.42,no tumor thrombus:2.77±0.31),necrosis(necrosis:2.36±0.42,no necrosis:2.79±0.39)and lymph node metastasis(metastasis:2.37±0.41,no metastasis:2.81±0.22,t=4.250,5.180,5.690,5.110,5.190,P<0.05).MiR-27b-3p was correlated to survival time(χ^2=5.080,P<0.05).Negative correlation was found between miR-27b-3p and IRF4(r=-0.600,P<0.05),and between miR-27b-3p and bcl-6(r=-0.690,P<0.05).Targeting relationship was found between miR-27b-3p and IRF4 by double luciferase reporter gene.than that in negative control group(t=3.240,4.330,P<0.05),the expression of IRF4 and bcl-6 was lower in miR-27b-3p transfection group.Co-transfection group could reverse the

关 键 词：结肠癌 微小RNA 干扰素调节因子4 增殖 预后 

分 类 号：R735.35[医药卫生—肿瘤]