食管鳞癌组织ADAMTS-5表达临床意义研究  

Expression of ADAMTS-5 and its clinical significance in esophageal squamous carcinoma

作　　者：盖领[1] 刘军[1] 袁心露[2] 姚宁华[3] 顾术东[1] 沈洪[1] 茅国新[1] 

机构地区：[1]南通大学附属医院化疗科,江苏南通226001 [2]南通大学附属医院内分泌科,江苏南通226001 [3]南通大学附属医院放疗科,江苏南通226001

出　　处：《中华肿瘤防治杂志》2017年第8期546-551,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：南通市级科技计划(MS22016071;MS22015126)

摘　　要：目的探讨含Ⅰ型血小板反应蛋白的解聚素金属蛋白酶5(a disintegrin and metalloprotease with thrombospondinmotifs 5,ADAMTS-5)蛋白在食管癌组织中的表达,对食管癌细胞增殖及迁移能力的影响,并分析其对患者预后影响的临床意义。方法选取正常食管上皮细胞株(HEEC)和4株食管鳞状细胞癌细胞株(TE1、TE10、TE11以及Eca109),采用蛋白质印迹法检测ADAMTS-5蛋白在细胞内的表达。通过RNA干扰技术在食管鳞状细胞癌细胞株TE10中转染ADAMTS-5-shRNA和Neg-shRNA后采用蛋白质印迹法检测ADAMTS-5蛋白的表达,四甲基偶氮唑蓝(MTT)法、划痕实验检测干扰ADAMTS-5表达对TE10细胞增殖以及迁移的影响。采用免疫组化法检测2009-04-15-2012-11-20南通大学附属医院97例食管鳞癌组织中ADAMTS-5蛋白的表达,分析其表达与患者临床病理特征及预后的关系。结果在食管鳞癌细胞株TE1、TE10、TE11和Eca109中ADAMTS-5蛋白相对表达量分别为2.10±0.06、5.70±0.21、4.50±0.32和4.60±0.39,明显高于食管正常上皮细胞株HEEC蛋白相对表达量(1.10±0.07),其中在TE10中的表达强度最高。使用小干扰技术转染TE10细胞株48h后,蛋白质印迹法检测结果显示,以GAPDH为内参照,未处理组、阴性对照组和干扰组的ADAMTS-5蛋白相对表达量分别为1.00±0.05、1.10±0.09和0.50±0.07,干扰组蛋白表达相比阴性对照组(P<0.001)和未处理组(P<0.001)明显下降;MTT法检测结果显示,转染72h后,未处理组、阴性对照组和干扰组的增殖率分别为(100.00±10.02)%、(98.04±9.80)%和(72.19±12.24)%,干扰组相比阴性对照组(P<0.001)和未处理组(P<0.001)细胞增殖明显抑制;在划痕24h后,干扰组迁移率为(46.34±1.27)%,而阴性对照组为(73.17±1.54)%,干扰组迁移能力明显抑制,t=40.33,P<0.001;ADAMTS-5蛋白表达在97例食管鳞癌组织中的阳性率为59.79%(58/97),明显高于癌旁组织的28.87%(28/97),与分化程度、淋巴结转移、临床分期和浸润深度�OBJECTIVE To detect the expression of ADAMTS5 and to investigate its effects on proliferation and migration within ESCC cell line as well as to analyze the significance of prognosis by Kaplan-Meier analysis.METHODS The expression of ADAMTS5 protein was detected by western blotting assay in human esophageal epithelial cell line and 4kinds of esophageal carcinoma cell lines,such as TE1,TE10,TE11 and Eca109.TE10 was transfected with ADAMTS5-shRNA to knockdown the expression of ADAMTS5.We detected the proliferation and migration of TE10 cells through MTT assay and cell wound scratch assay.The expression of ADAMTS5 was analyzed using immunohistochemistry on 97 separate cases of esophageal carcinoma paraffin-embedded slices.Pearsonχ~2 test was administrated to examine the association between ADAMTS5 and clinicopathological character.Kaplan-Meier analysis was used to evaluate the influence of the ADAMTS5 expression on prognosis of ESCC patients.RESULTS Western blotting showed that the expression of ADAMTS5 in esophageal squamous cell carcinoma cell lines such as TE1（2.10±0.06,P〈0.001）,TE10（5.70±0.21,P〈0.001）,TE11（4.50±0.32,P〈0.001）and Eca109（4.60±0.39,P〈0.001）were significantly higher than that in human esophageal epithelial cell line（1.10±0.07）,especially in TE10.The knockdown effectiveness of the ADAMTS5-shRNA was tested by western blotting.It was showed that the relative expression of ADAMTS5 protein in an untreated group,negative control group and interference group were 1.00±0.05,1.10±0.09 and 0.50±0.07respectively;although the expression of the interference group was significantly down regulation than the negative control（P〈0.001）and untreated（P〈0.001）groups.After TE10 cell line was transfected with shRNA for 72 h,MTT results showed that the proliferation rate in untreated,negative control and interference groups were（100.00±10.02）%,（98.04±9.80）% and（72.19±12.24）% respectively.With the negative control group（P〈0.001）and untreated group（P〈0.0

关 键 词：食管鳞癌 RNA干扰 含Ⅰ型血小板反应蛋白的解聚素金属蛋白酶 增殖 迁移 预后 

分 类 号：R735.1[医药卫生—肿瘤]