中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2015年
14期
2730-2734
,共5页
RNA 干扰%宫颈肿瘤%巨噬细胞移动抑制因子%上皮-间质转化%侵袭%转移
RNA 榦擾%宮頸腫瘤%巨噬細胞移動抑製因子%上皮-間質轉化%侵襲%轉移
RNA 간우%궁경종류%거서세포이동억제인자%상피-간질전화%침습%전이
RNA interference%Uterine cervical neoplasms%Macrophage migration inhibitory factor%Epithelial-mesenchymal transition%Invasion%Metastasis
目的：通过 RNA 干扰技术研究巨噬细胞移动抑制因子（MIF）对宫颈癌 Caski 细胞上皮-间质转化（EMT）的影响作用。方法通过构建重组真核表达载体 pGenesil-1-MIF siRNA，转染宫颈癌 Caski 细胞（Caski-pGenesil-MIF siRNA 组），同时设空载体转染组（Caski-pGenesil-1组）和空白对照组（Caski 组），采用荧光定量聚合酶链反应（PCR）和蛋白质印迹法（Western blot）检测 Caski 细胞中 MIF mRNA 相对表达量的变化，并检测转染前后 Caski 细胞中 EMT 相关指标E-cadherin 和 Vimentin 的表达变化。结果 pGenesil-1-MIF siRNA 转染 Caski 细胞后 MIF mRNA 的表达量减少，低于对照组，差异有统计学意义（P＜0.05）；RNA 干扰抑制 MIF 基因表达后，与对照组相比，pGenesil-1-MIF siRNA 组 Caski 细胞上皮标记物 E-cadherin mRNA 及蛋白表达水平上调（P＜0.05），间叶标记物 Vimentin mRNA 及蛋白表达水平显著下调（P＜0.05）。结论抑制 MIF基因表达可抑制宫颈癌 Caski 细胞发生 EMT 的能力。
目的：通過 RNA 榦擾技術研究巨噬細胞移動抑製因子（MIF）對宮頸癌 Caski 細胞上皮-間質轉化（EMT）的影響作用。方法通過構建重組真覈錶達載體 pGenesil-1-MIF siRNA，轉染宮頸癌 Caski 細胞（Caski-pGenesil-MIF siRNA 組），同時設空載體轉染組（Caski-pGenesil-1組）和空白對照組（Caski 組），採用熒光定量聚閤酶鏈反應（PCR）和蛋白質印跡法（Western blot）檢測 Caski 細胞中 MIF mRNA 相對錶達量的變化，併檢測轉染前後 Caski 細胞中 EMT 相關指標E-cadherin 和 Vimentin 的錶達變化。結果 pGenesil-1-MIF siRNA 轉染 Caski 細胞後 MIF mRNA 的錶達量減少，低于對照組，差異有統計學意義（P＜0.05）；RNA 榦擾抑製 MIF 基因錶達後，與對照組相比，pGenesil-1-MIF siRNA 組 Caski 細胞上皮標記物 E-cadherin mRNA 及蛋白錶達水平上調（P＜0.05），間葉標記物 Vimentin mRNA 及蛋白錶達水平顯著下調（P＜0.05）。結論抑製 MIF基因錶達可抑製宮頸癌 Caski 細胞髮生 EMT 的能力。
목적：통과 RNA 간우기술연구거서세포이동억제인자（MIF）대궁경암 Caski 세포상피-간질전화（EMT）적영향작용。방법통과구건중조진핵표체재체 pGenesil-1-MIF siRNA，전염궁경암 Caski 세포（Caski-pGenesil-MIF siRNA 조），동시설공재체전염조（Caski-pGenesil-1조）화공백대조조（Caski 조），채용형광정량취합매련반응（PCR）화단백질인적법（Western blot）검측 Caski 세포중 MIF mRNA 상대표체량적변화，병검측전염전후 Caski 세포중 EMT 상관지표E-cadherin 화 Vimentin 적표체변화。결과 pGenesil-1-MIF siRNA 전염 Caski 세포후 MIF mRNA 적표체량감소，저우대조조，차이유통계학의의（P＜0.05）；RNA 간우억제 MIF 기인표체후，여대조조상비，pGenesil-1-MIF siRNA 조 Caski 세포상피표기물 E-cadherin mRNA 급단백표체수평상조（P＜0.05），간협표기물 Vimentin mRNA 급단백표체수평현저하조（P＜0.05）。결론억제 MIF기인표체가억제궁경암 Caski 세포발생 EMT 적능력。
Objective To investigate the effect of RNA interference (RNAi) targeting macrophage migration inhibitory factor (MIF) on epithelial-mesenchymal transition (EMT) in human cervical carcinoma Caski cells. Methods Recombinant eukaryotic expression plasmid pGenesil-MIF siRNA was constructed and identified, then transfected into human cervical cancer cells Caski using a lipofectamine. Experimental cells were classified into 3 groups: Caski-pGenesil-MIF siRNA group, Caski-pGenesil-1 group and Caski group. Fluorescent quantitation polymerase chain reaction and Western blot were used to detect the the expression of MIF mRNA and the expression of EMT related markers such as E-cadherin and Vimentin in Caski cells was determined before and after transfection. Results The eukaryotic expression vector pGenesil-1-MIF siRNA significantly decreased the expression of mRNA of MIF in Caski cells (P<0.05). After RNA interference inhibited the expression of MIF gene in Caski cells, the expression of E-cadherin mRNA and protein in Caski-pGenesil-MIF siRNA group was significantly higher than those in control groups (P<0.05), while the expression of Vimentin in Caski-pGenesil-MIF siRNA group was significantly lower than those in control groups (P<0.05). Conclusion Inhibition of MIF gene expression in cervical cancer Caski cells can inhibit the ability of the occurrence of epithelial-mesenchymal transition.
