DAPPER1对食管鳞状细胞癌细胞恶性生物学行为的影响及其表观遗传学调节机制  

Effect of DAPPER1 on malignant biological behavior of esophageal carcinoma cells and its epigenetic regulatory mechanism

作　　者：郭艳丽[1] 邝钢[1] 周珍[1] 董稚明[1] 郭炜[1] 沈素朋[1] 梁佳[1] 郭鑫[1] 

机构地区：[1]河北医科大学第四医院河北省肿瘤研究所病理研究室,石家庄050011

出　　处：《临床与实验病理学杂志》2017年第8期827-831,共5页Chinese Journal of Clinical and Experimental Pathology

基　　金：国家自然科学基金(81472335);河北省医学科学研究重点课题计划项目(20170698)

摘　　要：目的检测DAPPER1蛋白对食管鳞状细胞癌(esophageal squamous cell cancer,ESCC)细胞株恶性生物学行为的影响,并进一步分析ESCC组织中DAPPER1蛋白表达及引起其表达异常的可能机制。方法应用MTT、克隆形成、划痕修复实验检测DAPPER1对ESCC细胞株恶性生物学行为的影响;应用RT-PCR及甲基化特异性PCR(methylation specific PCR,MSP)技术检测细胞株(TE13、T.Tn、Eca109)中DAPPER1 mRNA的表达及其启动子区甲基化状态;应用免疫组织化学(immunohistochemistry,IHC)法检测ESCC组织中DAPPER1蛋白的表达。结果 DAPPER1在3株细胞中呈弱表达或阴性,应用甲基化抑制剂5-氮杂-2'-脱氧胞苷(5-aza-2'-deoxycytidine,5-Aza-Dc)处理细胞后,其表达强度在各细胞株中均有不同程度的增加;同时,DAPPER1过表达及5-Aza-Dc处理可明显抑制TE13细胞的增殖及迁移能力;而应用组蛋白去乙酰化酶抑制剂trichostatin A(TSA)处理细胞株后,DAPPER1在各细胞株中的表达无明显改变;DAPPER1蛋白在ESCC组织中的表达较癌旁组织明显下调(P<0.01),且其表达与该基因启动子区域异常甲基化状态有关(P<0.01)。结论 ESCC中DAPPER1主要起抑癌基因作用,并且该基因启动子区域的异常高甲基化可能是引起其表达下调的主要机制之一。Purpose To investigate the effect of DAPPER1 on the malignant biological behavior in esophageal carcinoma cells, and further to analyze the expression and the possible reg- ulated mechanism of DAPPER1 in esophageal squamous cell cancer （ESCC） samples. Methods MTT assay, colony forma- tion assay and wound healing were used to examine the effect of DAPPER1 on malignant biological behavior in esophageal carci- noma cells, RT-PCR and MSP methods were applied respective- ly to examine the expression and the methylation status of DAP- PER1 in three ESCC cell lines （TE13, T. Tn, Eca109）. Immu- nohistochemistry was used to examine the DAPPER1 protein ex- pression. Results The negative or weak expression of DAP- PER1 was detected in ESCC cell lines. After treated with 5-Aza- 2＇-deoxycytidine （5-Aza-dC, a demethylation agent）, the ex-pression level of DAPPER1 was obviously increased. Mean- while, over expression of DAPPER1 or treated with 5-Aza-Dc could obviously inhibit proliferation and immigration abilities of TE13 cell line. The level of DAPPER1 expression had no obvi- ously change after treated with trichostatin A （TSA）. Decreased protein expression of DAPPERI was observed in ESCC tumor tis- sues compared with non-cancerous tissues （P 〈0. 01 ）, and as- sociated with the methylation status of this gene （ P 〈 0. 01 ）. Conclusion DAPPER1 plays an important role of tumor sup- pressor gene in esophageal carcinoma, and the aberrant hyperm- ethylation may be one of the main mechanisms in abnormal ex- pression of this gene.

关 键 词：食管肿瘤 甲基化 增殖 迁移 DAPPER1 

分 类 号：R735.1[医药卫生—肿瘤]