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目的:探讨微RNA-1250(microRNA-1250,miR-1250)及其宿主基因细胞凋亡关联酪氨酸激酶(apoptosis-associated tyrosine kinase,AATK)在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)发生和发展中的作用。方法:分别采用实时荧光定量PCR法和甲基化特异性PCR法检测AATK基因在食管癌细胞系和ESCC组织及其相应癌旁正常组织中的表达情况和甲基化状态,同时采用实时荧光定量PCR法检测miR-1250在ESCC组织及其相应癌旁正常组织中的表达情况。然后,分析miR-1250表达和AATK基因表达及其甲基化状态分别与ESCC组织病理学特征的关系,以及三者之间的相关性。结果:5-杂氮-2’-脱氧胞苷(5-aza-2’-deoxycytidine,5-Aza-dC)处理后,4株食管癌细胞系中AATK基因表达水平明显升高(P值均<0.05),并且其甲基化程度明显降低。在ESCC组织中,AATK mRNA平均表达量低于癌旁正常组织(P<0.05),AATK基因甲基化率明显高于癌旁正常组织(P<0.05);AATK mRNA相对表达量和AATK基因甲基化率均与淋巴结转移、组织分化程度及TNM分期密切相关(P值均<0.05),而与患者年龄和性别无关(P值均>0.05)。同时,ESCC组织中miR-1250相对表达量明显低于癌旁正常组织(P<0.05),并与淋巴结转移、组织分化程度及TNM分期密切相关(P值均<0.05),而与患者年龄和性别无关(P值均>0.05)。ESCC组织中miR-1250与AATK mRNA表达明显相关(P<0.05),而且miR-1250和AATK mRNA表达与AATK基因甲基化状态也均明显相关(P值均<0.05)。结论:ESCC的发生和发展可能与miR-1250和AATK低表达以及AATK基因高甲基化状态有关,而且miR-1250与AATK mRNA表达具有一致性。
Objective: To investigate the expression of microRNA-1250 (miR-1250) and its host gene apoptosis-associated tyrosine kinase (AATK) in esophageal squamous cell carcinoma ) The occurrence and development of the role. Methods: The expression and methylation status of AATK gene in esophageal cancer cell lines and ESCC tissues and corresponding normal tissues were detected by real-time fluorescence quantitative PCR and methylation-specific PCR respectively. Real-time fluorescence quantitative PCR method was used to detect the expression of miR-1250 in ESCC tissues and corresponding normal tissues. Then, the relationship between the expression of miR-1250, the expression of AATK gene and the methylation status of miR-1250 and the histopathological features of ESCC were analyzed, and the correlation between them was analyzed. Results: After treatment with 5-aza-2’-deoxycytidine (5-Aza-dC), the expression of AATK gene in four esophageal cancer cell lines was significantly increased <0.05), and the degree of methylation was significantly reduced. The average expression of AATK mRNA in ESCC tissues was lower than that in adjacent normal tissues (P <0.05), and the methylation rate of AATK gene was significantly higher than that in adjacent normal tissues (P <0.05). The relative expression of AATK mRNA and AATK gene A There was a close correlation between the rate of lymph node metastasis, histological differentiation and TNM stage (all P <0.05), but not with age and sex (P> 0.05). Meanwhile, the relative expression level of miR-1250 in ESCC was significantly lower than that in adjacent normal tissues (P <0.05), and closely correlated with lymph node metastasis, histological differentiation and TNM stage (all P <0.05) No gender (P> 0.05). The expression of miR-1250 and AATK mRNA was significantly correlated with ESCC (P <0.05), and the expression of miR-1250 and AATK mRNA was also correlated with the methylation status of AATK gene (all P <0.05). Conclusion: The occurrence and development of ESCC may be related to the low expression of miR-1250 and AATK and the hypermethylation status of AATK gene, and the consistency of miR-1250 and AATK mRNA expression.