miR-19a在食管鳞癌组织及细胞中的功能机制研究

Study on functional mechanism of miR-19a in esophageal squamous cell carcinoma

目的探讨miR-19a在食管鳞癌组织及细胞中的功能机制。方法收集30对新鲜食管鳞癌患者癌组织及其癌旁正常组织,根据病理分化程度及是否有淋巴结转移分别分为高-中分化组和中-低分化组,淋巴结转移组和未转移组,实时荧光定量(qRT-PCR)检测miR-19a的表达。在细胞水平上,干扰和过表达miR-19a后,通过四甲基偶氮唑盐微量酶反应比色法(MTT法)、细胞划痕、transwell试验检测miR-19a对食管鳞癌细胞系Eca109的增殖、迁移和侵袭的影响;流式细胞术检测转染miR-19a后对食管鳞癌细胞系Eca109周期和凋亡的影响。结果组织水平上,miR-19a在食管鳞癌组织中表达量(11.07±12.08),明显高于与其对应的癌旁组织(1.36±1.03),差异具有统计学意义。miR-19a在中-低分化组(6.92±3.61)和转移组(6.50±4.21)中的表达量也明显高于与其对应的高-中分化组(2.92±2.84)和未转移组(2.55±2.06),差异具有统计学意义。细胞水平上,在食管鳞癌Eca109细胞中,转染miR-19a Mimics促进了食管鳞癌细胞系Eca109的增殖、迁移和侵袭,流式细胞术结果显示转染miR-19a Mimics后促进食管鳞癌细胞进入S期,抑制细胞凋亡,而转染miR-19a Inhibitor抑制了食管鳞癌细胞系Eca109的增殖、迁移和侵袭,并将细胞阻滞在G2期,促进了细胞凋亡。结论miR-19a在食管鳞癌组织中高表达,并与分化程度和转移相关,促进了食管鳞癌细胞恶性表型的改变,有可能作为食管鳞癌临床诊治及精准诊疗的候选分子靶点。

Objective To investigate the functional mechanism of miR-19a in esophageal squamous cell carcinoma.Methods 30 pairs of fresh esophageal squamous cell carcinoma tissues and adjacent normal tissues were collected,according to the degree of pathological differentiation and whether there is lymph node metastasis,they are divided into high to medium differentiation group and medium to low differentiation group,lymph node metastasis group and non-metastasis group.The expression of miR-19a was detected by real-time quantitative fluorescence(qRT-PCR).After interference and overexpression of miR-19a at the cellular level,the effects of miR-19a on the proliferation,migration and invasion of esophageal cancer cell line Eca109 were detected by microenzyme reaction colorimetry(MTT),cell scratch and transwell assay.The effect of miR-19a transfection on the cycle and apoptosis of Eca109 cell line was detected by flow cytometry.Results At the tissue level,the expression level of miR-19a in esophageal squamous cell carcinoma tissues(11.07±12.08)was significantly higher than that in adjacent tissues(1.36±1.03),and the difference was statistically significant.Moreover,the expression level of miR-19a in the medium to low differentiation group(6.92±3.61)and metastasis group(6.50±4.21)was significantly higher than that in the corresponding high to medium differentiation group(2.92±2.84)and non-metastasis group(2.55±2.06),and the difference was statistically significant.At the cellular level,in esophageal cancer Eca109 cells,transfection of miR-19a Mimics promoted the proliferation,migration and infiltration of esophageal cancer cell line Eca109.Flow cytometry results showed that transfection of miR-19a Mimics promoted esophageal cancer cells to enter the S phase and inhibited apoptosis.However,transfection with miR-19a inhibitor inhibited the proliferation,migration and invasion of the Eca109 cell line,and blocked the cells in G2 phase,promoting cell apoptosis.Conclusion miR-19a is highly expressed in esophageal cancer tissues and is related to differentiation and metastasis,which promotes the change of malignant phenotype of esophageal cancer cells,and it may be a candidate molecular target for clinical diagnosis and treatment of esophageal cancer.