黏蛋白1在食管鳞癌上皮细胞-间质细胞转化过程中的作用  

The effect of mucin 1 in epithelial-mesenchymal transition of esophageal squamous cell carcinoma

作　　者：何晓鹏[1] 刘曦[2] 孙智[1] 史墨[1] 陈钢[1] 姜文鹏[1] 王洲 刘相燕[1] 

机构地区：[1]山东大学附属省立医院胸外科,济南250021 [2]山东大学医学院2011级临床医学系

出　　处：《中华实验外科杂志》2013年第8期1623-1625,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81272352）;山东省科学技术发展计划资助项目（2011YD19009）;山东省医药卫生科技发展计划资助项目（2011HD007）

摘　　要：目的观察黏蛋白1（MUC1）对食管鳞癌细胞生物学行为的影响并探讨其在上皮细胞一间质细胞转化过程中的作用。方法设计构建靶向MUC1基因的短发卡RNA（shRNA）慢病毒载体pGC-LV-MUC1小干扰RNA（siRNA）并感染人食管癌细胞株Eca109，检测其对MUC1表达的抑制作用；应用噻唑蓝（M1Tr）与Transwell侵袭实验检测抑制MUC1对肿瘤细胞增殖、侵袭能力的影响，并采用实时定量逆转录聚合酶链反应（RT-qPCR）与Westernblot检测E-钙黏蛋白（E-cadherin）在各组细胞中的表达差异。结果与对照组比较，感染pGC-LV-MUC1siRNA后，Eca109细胞内MUC1基因水平（1．000±0．061比0．612±0．019）和蛋白水平（11．230±1．120比7．440±0．970）明显降低（P〈0．05）；感染pGC-LV-MUC1siRNA能显著抑制Eca109细胞的增殖活性，48h（0．393±0．028比0．281±0．020），72h（0．792±0．052比0．463±0．028，P〈0．05）；感染pGC-LV-MUC1siRNA后，Eca109细胞的侵袭数量也明显降低[（87±4）个比（48±7）个，P〈0．05]；而Eca109细胞内E．cadherin基因水平（1．000±0．072比2．510±0．767）和蛋白水平（0．070±0．020比0．130±0．030）明显增高（P〈0．05）。结论MUC1可能通过诱导上皮细胞-间质细胞转化（EMT）而促进食管鳞癌细胞增殖、侵袭。Objective To detect the effect of gene silence targeting mucin 1 （MUC1） on biologi- cal behaviour of esophageal squamous cell carcinoma, and explore the function of MUC1 in the process of epithehal-mesenchymal transition. Methods To construct the short hairpin RNA （shRNA） lentiviral vec- tor targeting MUC1. After transfection, the interference effect of MUC1 on Eca109 cells was detected by re- al-time reverse transcriptase-polymerase chain reaction （RT-qPCR） and Western blotting. The cell prolif- eration of Eca109 was assessed by methyl thiazol tetrazolium （MTT） and the cell invasion was detected by transwell assay. Finally, RT-PCR and Western blot were used to examine the expression levels of E-cadherin in Eca109 cells. Results Compared with the untransfected control, MUC1 expression in Eca109 cells trans- fected with MUC1 shRNA was decreased significantly at both mRNA （ 1. 000 ±~ 0. 061 vs. 0. 612 ± 0. 019 ） and protein levels （ 11. 230 ± 1. 120 vs. 7. 440 ± 0. 970） （P 〈 0. 05 ）. Meanwhile, the cell proliferation of Eca109 ceils transfected with MUC1 shRNA was significantly lower at 48 h （0. 393 ± 0. 028 vs. 0. 281 ±0. 020）and 72 h （0. 792± 0. 052 vs. 0. 463 ±0. 028）, respectively （P all 〈0. 05）, and the invasion a- bility of which was decreased obviously （87 ± 4 vs. 48± 7, P 〈 0. 05 ）. Notable, E-cadherin expression in Eca109 cells transfected with MUC1 shRNA was significantly higher at both mRNA （ 1. 000 ± 0. 072 vs. 2.510±0.767） andprotein levels （0.070±0.020 vs. 0.130±0.030, P〈0.05）. Conclusion MUC1 might promote the invasion and metastasis of esophageal squamous cell carcinoma by the induction of epi- thelial-mesenehymal transition. [ Key words ]

关 键 词：食管鳞癌 上皮细胞-间质细胞转化 肿瘤侵袭转移 黏蛋白1 

分 类 号：R735.1[医药卫生—肿瘤]