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目的:通过检测食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)中长链非编码RNA XLOC_002319(long non-colding RNA XLOC_002319,lncRNA XLOC_002319)基因的表达及其甲基化状态,探讨XLOC_002319基因在ESCC发生及发展中的作用。方法:分别应用RT-PCR以及甲基化特异性PCR(methylation specific PCR,MSP)的方法检测DNA甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷(5-aza-2’-deoxycitydine,5-aza-d C)处理前后的食管癌细胞株(TE1、TE13、Yes-2、Eca109和T.TN)、ESCC组织以及癌旁正常组织、食管上皮内瘤变(esophageal intraepithelial neoplasia,EIN)组织中XLOC_002319基因的表达和甲基化状态。结果:未经5-aza-d C处理的5种食管癌细胞中XLOC_002319基因的表达均呈阴性或弱阳性,经5-azad C的5种食管癌细胞中XLOC_002319基因的表达均增高。5株食管癌细胞在5-aza-d C处理前表现为XLOC_002319高甲基化状态,处理后,Eca109和T.TN细胞系中XLOC_002319基因甲基化程度降低,其余3株细胞系中XLOC_002319基因均表现为非甲基化状态。XLOC_002319基因在ESCC组织中的表达显著低于食管上皮内瘤变组织和癌旁正常组织(P<0.01),并与组织学分化程度和TNM分期密切相关(P<0.05)。ESCC组织中XLOC_002319基因启动子区甲基化率为63.75%(51/80),显著高于食管上皮内瘤变组织和癌旁正常组织(P<0.01),并与淋巴结转移、组织学分化程度和TNM分期密切相关(P<0.05)。发生XLOC_002319基因甲基化的ESCC组织中XLOC_002319基因表达显著低于未发生甲基化的ESCC组织(P<0.01)。结论:XLOC_002319基因在ESCC中的低表达可能与ESCC的发生密切相关,且其启动子区甲基化可能是导致其表达沉默的机制之一。
OBJECTIVE: To investigate the expression of XLOC_002319 and its methylation status in esophageal squamous cell carcinoma (ESCC) Occurrence and development of the role. Methods: RT-PCR and methylation-specific PCR (MSP) were used to detect the DNA methylation transferase inhibitor 5-aza-2’-deoxycytidine (5-aza- (TE1, TE13, Yes-2, Eca109 and T.TN), ESCC tissues and adjacent normal tissues, esophageal intraepithelial neoplasia (esophageal neoplasia) before and after treatment with 5-aza- , EIN) tissue XLOC_002319 gene expression and methylation status. Results: The expression of XLOC_002319 gene in 5 esophageal cancer cells without 5-aza-d C was negative or weakly positive. The expression of XLOC_002319 gene in 5 esophageal cancer cells with 5-azad C was increased. Five esophageal cancer cells showed hypermethylation status before treatment with 5-aza-d C, and the methylation level of XLOC_002319 in Eca109 and T.TN cell lines decreased after treatment. XLOC_002319 genes in the other three lines all showed Unmethylated status. The expression of XLOC_002319 in ESCC was significantly lower than that in esophageal intraepithelial neoplasia and adjacent normal tissues (P <0.01), which was closely related to the histological differentiation and TNM stage (P <0.05). The methylation rate of XLOC_002319 gene promoter region in ESCC was 63.75% (51/80), which was significantly higher than that in esophageal intraepithelial neoplasia and adjacent normal tissue (P <0.01), and was correlated with lymph node metastasis, histological differentiation And TNM stage is closely related (P <0.05). The gene expression of XLOC_002319 in ESCC tissues with XLOC_002319 gene methylation was significantly lower than that in ESCC tissues without methylation (P <0.01). Conclusion: The low expression of XLOC_002319 in ESCC may be closely related to the occurrence of ESCC, and its promoter methylation may be one of the mechanisms leading to its silence.