摘要
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 and methylation status of long non-coding RNA XLOC_002319 (lncRNA XLOC_002319) in esophageal squamous cell carcinoma (ESCC), and to elucidate its role in the progression of ESCC. Methods: Reverse transcription polymerase chain reaction (RT-PCR) and methylation specific PCR (MSP) were used respectively to detect the expression and methylation status of XLOC_002319 in esophageal cancer cell lines (TE1, TE13, Yes-2, Eca109, T.TN) treated or untreated with DNA methyltransferase inhibitor 5-aza-2′-detoxycytidine (5-aza-dC), and in cancer-adjacent normal tissues, esophageal intraepithelial neoplasia (EIN) tissues and ESCC tissues. Results:Negative or weak positive expression of XLOC_002319 was detected in the five esophageal cancer cell lines untreated with 5-aza-dC; however, after the treatment with 5-aza-dC, the expression of XLOC_002319 was enhanced. The methylation status of XLOC_002319 was highly expressed in esophageal cancer cell lines before the treatment of 5-aza-dc, however, after the treatment, the methylation level was decreased in Eca109 and T.TN cell lines, while the status in other three cell lines was negative. The expression of XLOC_002319 gene in ESCC tissues was significantly reduced compared to cancer-adjacent tissues and EIN tissues (P〈0.01), and was closely associated with pathological differentiation and TNM stage (P〈0.05). The methylation frequency of XLOC_002319promoter in ESCC tissues(63.75% /[51/80/]) was significantly higher than that in EIN tissues and adjacent normal tissues (P〈0.01), and it was also associated with lymph node metastasis, pathological differentiation and TNM stage (P〈0.05). The expressions of XLOC_002319 in ESCC tumor tissues with methylation of the gene was significantly lower than that in tumor tissues with unmethylation of the gene (P〈0.01). Conclusion: Aberrant low expression of lncRNA XLOC_002319 was closely related to the development and occurrence of ESCC, and promoter methylation might be one of the mechanisms for inactivation of XLOC_002319 in ESCC.
作者
梁佳
邝钢
刘胜男
郭炜
沈素朋
丁春艳
李慧杰
董稚明
LIANG Jia KUANG Gang LIU Shengnan GUO Wei SHEN Supeng DING Chunyan LI Huijie DONG Zhiming(Cancer Institute of Hebei Province, the Fourth Hospital Affiliated to Hebei Medical University, Shijiazhuang 050011, Hebei, China Department of Oncology, Shijiazhuang City First People's Hospital, Shijiazhuang 050011, Hebei, China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2016年第5期675-681,共7页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.81572441)
河北省自然科学基金资助项目(No.H2015206196)~~