摘要
目的探讨微小RNA-3651(miR-3651)与食管癌侵袭进展的关系及分子机制。方法应用肿瘤基因组图谱(TCGA)数据库检索食管癌相关的miRNA。实时定量聚合酶链反应(Real-time PCR)法检测30例临床新鲜食管癌组织、癌旁正常食管黏膜miR-3651表达;同时检测食管癌细胞株Eca-109、TE-10及食管上皮细胞株HET-1A中miR-3651表达,对数据库检索结果进行验证。用miR-3651抑制物(inhibitors)转染Eca-109细胞,检测转染对Eca-109细胞活性及迁移、侵袭能力的影响;同时检测抑制miR-3651后Eca-109细胞迁移侵袭相关基因表达情况。结果TCGA数据库结果显示miR-3651在食管癌表达(10.332±18.538)高于正常组织(2.923±2.216,Z=2.913,P<0.01);高表达miR-3651是患者预后差的标志(χ^(2)=5.112,P<0.05)。验证结果显示,食管癌组织中miR-3651水平(4.474±0.790)明显高于正常食管黏膜(1.005±0.241,t=23.005,P<0.01)。Eca-109、TE-10细胞miR-3651水平(1.107±0.093、0.823±0.121)明显高于HET-1A(0.276±0.050,F=62.413,P<0.01)。miR-3651 inhibitors转染后Eca-109细胞活性明显低于空白组、对照组(F=194.228,P<0.01);转染组细胞迁移侵袭能力明显低于空白组、对照组(F=100.881,P<0.01;F=136.652,P<0.01)。miR-3651 inhibitors组Eca-109细胞MMP-9、ICAM-1表达明显低于空白组和对照组[mRNA:(0.399±0.036)、(1.087±0.073)、(1.061±0.092),F=79.330,P<0.01;(0.437±0.052)、(1.002±0.086)、(1.123±0.111),F=35.742,P<0.01.蛋白:(0.446±0.116)、(1.034±0.246)、(1.047±0.227),F=8.471,P=0.018;(0.384±0.109)、(1.311±0.297)、(1.332±0.302),F=13.783,P=0.006],而TIMP-1的表达明显高于空白组和对照组[mRNA:(2.788±0.269)、(1.178±0.164)、(1.301±0.099),F=44.326,P<0.01.蛋白:(0.841±0.191)、(0.354±0.120)、(0.328±0.076),F=13.291,P<0.01]。结论miR-3651可能通过调控一些迁移侵袭相关基因的表达而促进食管癌进展转移,检测miR-3651表达有助于判断食管癌预后。
Objective To investigate the relationship between microRNA-3651(miR-3651)and the invasion and progression of esophageal cancer and its molecular mechanism.Methods The TCGA database was used to search for miRNAs related to esophageal cancer.Real-time quantitative PCR(real time PCR)method was used to detect the expression of miR-3651 in 30 cases of clinical fresh esophageal cancer tissues and normal esophageal mucosa adjacent to the cancer.Simultaneously the miR-3651 expression was detected in esophageal cancer cell lines Eca-109,TE-10 and esophageal epithelial cell line HET-1A,to verify the database search results.Eca-109 cells were transfected with miR-3651 inhibitors to detect the effect of transfection on Eca-109 cell viability and migration and invasion ability.Also the expression of genes related to migration and invasion of Eca-109 cells after miR-3651 inhibition was detected.Results The results of the TCGA database showed that miR-3651 expression was significantly higher in esophageal cancer(10.332±18.538)than in normal tissue(2.923±2.216,Z=2.913,P<0.01).Highly expressed miR-3651 was a sign of poor prognosis(χ^(2)=5.112,P<0.05).The verification results showed that the level of miR-3651 in esophageal cancer tissue(4.474±0.790)was significantly higher than that in normal esophageal mucosa(1.005±0.241,t=23.005,P<0.01).The level of miR-3651 in Eca-109(1.107±0.093)and TE-10 cells(0.823±0.121)was significantly higher than that in HET-1A cells(0.276±0.050,F=62.413,P<0.01).Eca-109 cell activity was significantly weaker in miR-3651 inhibitors group than in blank group and control group(F=194.228,P<0.01),and migration and invasion ability in miR-3651 inhibitors group was significantly reduced as compared with that in blank group and control group(F=100.881,136.652,P<0.01).The expression levels of matrix metalloproteinase(MMP)-9 and intercellular adhesion molecule-1(ICAM-1)in Eca-109 cells were significantly reduced in miR-3651 inhibitors group as compared with those in blank group and control group[mRNA:(0.39
作者
李淑军
姚继方
江蒲
Li Shujun;Yao Jifang;Jiang Pu(Operating Theatre,Fourth Hospital of Hebei Medical University,Shijiazhuang 050019,China;Department of Thoracic Surgery,Fourth Hospital of Hebei Medical University,Shijiazhuang 050019,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第8期1455-1458,共4页
Chinese Journal of Experimental Surgery
关键词
食管癌
微小RNA
侵袭
生物信息学
预后
Esophageal cancer
MicroRNA 3651
Invasion
Bioinformatics analysis
Prognosis
