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目的:检测食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)中长链非编码RNA XLOC_008370(long non-colding RNA XLOC_008370,lnc RNA XLOC_008370)的表达及其甲基化状态,探讨XLOC_008370在ESCC发生及发展中的作用机制。方法:分别应用RT-PCR以及甲基化特异性PCR(methylation specific PCR,MSP)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ESCC细胞系(TE1、TE13、Eca109、T.TN、YES-2)以及ESCC组织及相应癌旁正常组织中XLOC_008370的表达和甲基化状态,分析其与临床病理特征的关系。结果:5种ESCC细胞中XLOC_008370的表达均呈阴性或弱阳性,经5-Aza-d C处理后,5种细胞中XLOC_008370的表达均升高。5种细胞中XLOC_008370基因呈高甲基化状态,应用5-Aza-d C处理后,Eca109、Yes-2细胞中XLOC_008370基因甲基化程度降低,非甲基化程度增加,其余3种细胞中XLOC_008370基因均表现为非甲基化状态。XLOC_008370在ESCC组织中的表达显著低于癌旁正常组织(P<0.05),并与TNM分期和组织学分化程度密切相关(P<0.05)。ESCC组织中XLOC_008370的启动子区甲基化率为(54.02%,47/87),显著高于癌旁正常组织(9.20%,8/87)(P<0.05),并与TNM分期和组织学分化程度密切相关(P<0.05)。发生XLOC_008370甲基化的ESCC组织中XLOC_008370的表达显著低于未发生甲基化的ESCC组织(P<0.05)。结论:XLOC_008370的异常低表达可能与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 detect the expression and methylation status of long non-coding RNA XLOC_008370 (long non-colding RNA XLOC_008370, lnc RNA XLOC_008370) in esophageal squamous cell carcinoma (ESCC) and to explore the role of XLOC_008370 in ESCC development and The mechanism of development. Methods: RT-PCR and methylation specific PCR (MSP) were used to detect the DNA methyltransferase inhibitor 5-Aza-2’-deoxycytidine (5-Aza-2’- (TE1, TE13, Eca109, T.TN, YES-2) before and after treatment with deoxycitydine and 5-Aza-d C and the expression and methylation status of XLOC_008370 in ESCC tissues and corresponding paracancerous normal tissues. Its relationship with clinicopathological features. Results: The expression of XLOC_008370 in all 5 ESCC cells was negative or weakly positive. The expression of XLOC_008370 in 5 kinds of ESCC cells was increased after 5-Aza-d C treatment. XLOC_008370 gene was hypermethylated in 5 kinds of cells. After 5-Aza-d C treatment, the methylation level of XLOC_008370 gene in Eca109 and Yes-2 cells decreased and the degree of non-methylation increased. In the other three kinds of cells, XLOC_008370 All showed non-methylation status. The expression of XLOC_008370 in ESCC tissues was significantly lower than that in adjacent normal tissues (P <0.05), and was closely related to TNM staging and histological differentiation (P <0.05). The methylation rate of XLOC_008370 in ESCC was 54.02%, 47/87, which was significantly higher than that in normal tissue (9.20%, 8/87) (P <0.05), and correlated with TNM staging and histological score Degree of closeness (P <0.05). The XLOC_008370 expression in XLOC_008370 methylated ESCC tissues was significantly lower than that in the non-methylated ESCC tissues (P <0.05). Conclusion: The abnormally low expression of XLOC_008370 may be closely related to the occurrence of ESCC, and its promoter methylation may be one of the mechanisms leading to its silence.