中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
8期
1623-1625
,共3页
何晓鹏%刘曦%孙智%史墨%陈钢%姜文鹏%王洲%刘相燕
何曉鵬%劉晞%孫智%史墨%陳鋼%薑文鵬%王洲%劉相燕
하효붕%류희%손지%사묵%진강%강문붕%왕주%류상연
食管鳞癌%上皮细胞-间质细胞转化%肿瘤侵袭转移%黏蛋白1
食管鱗癌%上皮細胞-間質細胞轉化%腫瘤侵襲轉移%黏蛋白1
식관린암%상피세포-간질세포전화%종류침습전이%점단백1
Esophageal squamous cell carcinoma%Epithelial-mesenchymal transition%Tumor invasion and metastasis%Mucin 1
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-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(MTT)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Western blot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-MUC1 siRNA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-MUC1 siRNA能显著抑制Eca109细胞的增殖活性,48 h (0.393±0.028比0.281 ±0.020),72 h(0.792±0.052比0.463 ±0.028,P＜ 0.05)；感染pGC-LV-MUC1 siRNA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.
目的 觀察黏蛋白1(MUC1)對食管鱗癌細胞生物學行為的影響併探討其在上皮細胞-間質細胞轉化過程中的作用.方法 設計構建靶嚮MUC1基因的短髮卡RNA(shRNA)慢病毒載體pGC-LV-MUC1小榦擾RNA(siRNA)併感染人食管癌細胞株Eca109,檢測其對MUC1錶達的抑製作用；應用噻唑藍(MTT)與Transwell侵襲實驗檢測抑製MUC1對腫瘤細胞增殖、侵襲能力的影響,併採用實時定量逆轉錄聚閤酶鏈反應(RT-qPCR)與Western blot檢測E-鈣黏蛋白(E-cadherin)在各組細胞中的錶達差異.結果 與對照組比較,感染pGC-LV-MUC1 siRNA後,Eca109細胞內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)明顯降低(P＜0.05)；感染pGC-LV-MUC1 siRNA能顯著抑製Eca109細胞的增殖活性,48 h (0.393±0.028比0.281 ±0.020),72 h(0.792±0.052比0.463 ±0.028,P＜ 0.05)；感染pGC-LV-MUC1 siRNA後,Eca109細胞的侵襲數量也明顯降低[(87±4)箇比(48±7)箇,P＜0.05]；而Eca109細胞內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)明顯增高(P＜0.05).結論 MUC1可能通過誘導上皮細胞-間質細胞轉化(EMT)而促進食管鱗癌細胞增殖、侵襲.
목적 관찰점단백1(MUC1)대식관린암세포생물학행위적영향병탐토기재상피세포-간질세포전화과정중적작용.방법 설계구건파향MUC1기인적단발잡RNA(shRNA)만병독재체pGC-LV-MUC1소간우RNA(siRNA)병감염인식관암세포주Eca109,검측기대MUC1표체적억제작용；응용새서람(MTT)여Transwell침습실험검측억제MUC1대종류세포증식、침습능력적영향,병채용실시정량역전록취합매련반응(RT-qPCR)여Western blot검측E-개점단백(E-cadherin)재각조세포중적표체차이.결과 여대조조비교,감염pGC-LV-MUC1 siRNA후,Eca109세포내MUC1기인수평(1.000±0.061비0.612±0.019)화단백수평(11.230±1.120비7.440±0.970)명현강저(P＜0.05)；감염pGC-LV-MUC1 siRNA능현저억제Eca109세포적증식활성,48 h (0.393±0.028비0.281 ±0.020),72 h(0.792±0.052비0.463 ±0.028,P＜ 0.05)；감염pGC-LV-MUC1 siRNA후,Eca109세포적침습수량야명현강저[(87±4)개비(48±7)개,P＜0.05]；이Eca109세포내E-cadherin기인수평(1.000±0.072비2.510±0.767)화단백수평(0.070±0.020비0.130±0.030)명현증고(P＜0.05).결론 MUC1가능통과유도상피세포-간질세포전화(EMT)이촉진식관린암세포증식、침습.
Objective To detect the effect of gene silence targeting mucin 1 (MUC1) on biological behaviour of esophageal squamous cell carcinoma,and explore the function of MUC1 in the process of epithelial-mesenchymal transition.Methods To construct the short hairpin RNA (shRNA) lentiviral vector targeting MUC1.After transfection,the interference effect of MUC1 on Eca109 cells was detected by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blotting.The cell proliferation of Ecal09 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 Ecal09 cells.Results Compared with the untransfected control,MUC1 expression in Eca109 cells transfected 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 cells 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 ability 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) and protein 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 epithelial-mesenchymal transition.
