miR-26a在食管癌中的表达及其对食管癌细胞增殖、侵袭能力的分析研究

Expression and clinical value of miR-26a in esophageal squamous-cell carcinoma

目的探究食管癌组织中微小RNA(miR)-26a表达水平变化及其临床意义,研究分析其对食管癌细胞(TE-1细胞和Eca109细胞)增殖、侵袭能力的影响。方法选取2014年1月至2018年12月在江苏大学附属宜兴人民医院手术切除并经病理学检查确诊的食管癌标本45例,采用实时荧光定量PCR(RT-PCR)检测食管癌组织及癌旁正常组织中miR-26a水平,分析miR-26a水平与临床病理特征的关系。选用人ESCC细胞系(TE-1、Eca109)及正常人食管鳞状上皮细胞系(HEEC)作为研究对象,分为对照组(WT组)、空载体转染组(NC组)和miR-26a组(miR-26a组)。其中对照组不作处理;空载体转染组对细胞进行空载体转染;miR-26a组对细胞进行miR-26a质粒转染。Cell Counting Kit-8法检测细胞活性;Annexin V-FITC PI法检测细胞凋亡情况。标准程序用流式细胞仪检测,结果用细胞周期拟合软件分析细胞周期。集落形成实验检测细胞集落形成。结果食管癌组织中miR-26a水平低于癌旁组织(P<0.05)。miR-26a水平与食管癌肿瘤大小、肿瘤细胞分化不良、肿瘤浸润及局部淋巴结转移数量相关(P<0.05),而与年龄、性别无关(P>0.05)。转染miR-26a组TE-1细胞和Eca109细胞活力下降、集落形成减少、处于S期(DNA合成期)和G2/M期(有丝分裂期)的细胞显著减少,细胞凋亡增加。结论食管癌组织和细胞中,miR-26a的表达显著减少;在ESCC细胞(TE-1和Eca109)中过表达miR-26a,可抑制细胞的增殖,降低细胞活性,并促进其凋亡。miR-26a有望成为食管癌诊疗新的靶点。

【Objective】To study the expression of microRNA-26a in esophageal squamous-cell carcinoma and ESCC cells,and to analyze its effects on cell proliferation and invasion.【Methods】From January 2014 to December 2018,45 patients with esophageal squamous-cell carcinoma who were diagnosed with surgical resection by pathological examination in our hospital were selected as the subjects.Real-time quantitative PCR(RT-PCR)was used to examine miR-26a expression in 45 ESCC tissues and paired adjacent non-cancerous tissues.ESCC cells(Ecal09,TE-1)and normal esophageal epithelial cell HEEC were selected and divided into three groups:mock group,control group and MIR-26a mimic group.After transfection,the expression of miR-26a at mRNA level was detected by quantitative real-time PCR.Cell viability was detected by CCK-8 and the percentage of apoptosis was detected by flow cytomemc analysis.【Results】The expression levels of miR-26a in ESCC cancer tissues were significantly higher than those in the surrounding tissues.MiR-26a was significantly associated with tumor grade(P=0.006)and tumor size(P=0.016),nodal status(P=0.010),tumor invasion(P=0.038)but not with age,gender.Relative levels of miR-26a in ESCC cells(Ecal09,TE-l)were evidently lower than those in normal esophageal epithelial cell HEEC.MiR-26a mimic significantly suppressed cell proliferation and invasion(cell viability,cell apoptosis rate,cell cycle).【Conclusion】The expression quantity of miR-26a is related to invasion and appreciation ability,significantly associated with tumor grade and tumor size,nodal status,tumor invasion;miR-26a plays tumor suppressive role in ESCC,and may be a potential target for the diagnosis and therapy of ESCC.