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作 者:孙玉芳[1] 龙翔[2] 张小强[2] 魏益平[2] 徐建军[2] 吴永兵[2]
机构地区:[1]南昌大学第四附属医院,330003 [2]南昌大学第二附属医院胸心外科
出 处:《中华实验外科杂志》2014年第10期2197-2199,F0003,F0004,共5页Chinese Journal of Experimental Surgery
基 金:江西省教育厅科技研究项目(GJJ13170)
摘 要:目的 探讨四跨膜蛋白CD81表达和食管癌细胞侵袭转移的关系.方法 采用实时定量逆转录聚合酶链反应(RT-qPCR)及Western blot检测正常食管上皮细胞与3株不同转移潜能的食管癌细胞中CD81的表达;免疫组织化学检测12例食管癌及相应癌旁组织中CD81表达;慢病毒干扰CD81表达,研究CD81对食管癌细胞侵袭的影响;并研究CD81表达与E-钙黏蛋白(E-cadherin)表达的关系;最后通过免疫组织化学法检测食管癌组织中CD81与E-cadherin表达,以进一步验证.结果 CD81在食管癌细胞中表达明显高于正常食管上皮细胞(mRNA:1.87±0.09比1.25 ±0.05;蛋白:1.53 ±0.06比0.39±0.02),在食管癌细胞KYSE150、KYSE450、KYSE510中CD81 mRNA表达分别为2.40 ±0.11、2.10±0.08与1.10 ±0.09,蛋白表达为1.73 ±0.10、2.21 ±0.07与0.64±0.01;免疫组织化学检测显示12例癌组织中表达明显均高于癌旁组.食管癌细胞中干扰CD81表达后,细胞的侵袭能力下调(121 ±23比45 ±19;152 ±48比75 ±37),E-cadherin表达上调.食管癌组织中高表达CD81伴E-cadherin表达下调.结论 高表达四跨膜蛋白CD81可能通过下调E-cadherin蛋白表达促进食管癌侵袭.Objective To investigate the role of tetraspanin CD81 in the invasion and metastasis of esophageal cancer (ES) cells.Methods The expression of tetraspanin CD81 mRNA and protein was detected by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blotting in the normal esophageal epithelial cells (HEEC) and three ES cell lines with differential potentials of metastasis and invasion.The expression of CD81 in 12 cases of ES and their corresponding para-carcinoma tissues was detected by immunohistochemistry (IHC).By CD81 RNA interference,the role of CD81 in ES cell invasion and E-cadherin expression ws studied.Finally,the relationship between the expression of CD81 and E-cadherin was also investigated.Results RT-qPCR and Western blotting showed that the CD81 expression was higher in ES cells than in HEEC (mRNA:1.87 ±0.09 vs.1.25 ±0.05; protein:1.53 ±0.06 vs.0.39 ±0.02),and the expression of CD81 mRNA in KSYE450,KYSE150 and KYSE510 was 2.40 ±0.11,2.10 ±0.08 and 1.10 ±0.09,and that of protein was 1.73 ±0.10,2.21 ±0.07 and 0.64 ± 0.01,respectively.The IHC showed CD81 expression was higher in 12 cases of ES tissues than in the corresponding para-carcinoma tissues.The down-regulation of CD81 in ES cells impaired the cells invasion (121 ±23 vs.45 ± 19; 152 ±48 vs.75 ±37) and E-cadherin expression.The IHC showed that the CD81 overexpression was associated with low expression of E-cadherin in ES tissues.Conclusion Overexpression of CD81 promotes the invasion of ES by down-regulation of E-cadherin expression.
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