CAJ | 학술논문

目的观察甘露糖敏感性绿脓杆菌制剂(PA-MSHA)对肝细胞肝癌上皮-间充质转化(EMT)的作用.方法培养人肝癌细胞株MHCC97L及HepG2,以不同剂量的PA-MSHA作用于肝癌细胞,显微镜观察细胞形态学变化,免疫荧光技术及Western blot检测PA-MSHA作用前后肝癌细胞EMT相关基因及转录因子的表达,免疫组织化学及反转录-聚合酶链反应(RT-PCR)技术检测PA-MSHA治疗前后裸鼠模型肝癌组织中钙黏蛋白(E-cadherin)及转录因子Snail和Slug表达.结果 Western blot结果显示与对照组比较PA-MSHA治疗组肝癌细胞E-cadherin表达显著增强,而波形蛋白(Vimentin)、纤维粘连蛋白(Fibronecin)、核因子-kappa B(NF-κB)以及转录因子Snail和Slug的表达显著降低,其效应呈剂量依赖性,而外源性甘露糖显著抑制PA-MSHA对上述蛋白的调控作用.免疫荧光也显示PA-MSHA上调肝癌细胞E-cadherin表达而抑制Vimentin的表达,显微镜下细胞形态也由不规则的梭形转变为相对规则的卵圆形.免疫组织化学结果显示荷瘤裸鼠经PA-MSHA治疗后肝癌组织E-cadherin的表达显著增强,而Snail及Slug的转录则受到显著抑制(P＜0.05).结论 PA-MSHA可以通过识别结合肝癌细胞表面的甘露糖显著抑制肝癌EMT的发生.
목적 관찰감로당민감성록농간균제제(PA-MSHA)대간세포간암상피-간충질전화(EMT)적작용.방법 배양인간암세포주MHCC97L급HepG2,이불동제량적PA-MSHA작용우간암세포,현미경관찰세포형태학변화,면역형광기술급Western blot검측PA-MSHA작용전후간암세포EMT상관기인급전록인자적표체,면역조직화학급반전록-취합매련반응(RT-PCR)기술검측PA-MSHA치료전후라서모형간암조직중개점단백(E-cadherin)급전록인자Snail화Slug표체.결과 Western blot결과현시여대조조비교PA-MSHA치료조간암세포E-cadherin표체현저증강,이파형단백(Vimentin)、섬유점련단백(Fibronecin)、핵인자-kappa B(NF-κB)이급전록인자Snail화Slug적표체현저강저,기효응정제량의뢰성,이외원성감로당현저억제PA-MSHA대상술단백적조공작용.면역형광야현시PA-MSHA상조간암세포E-cadherin표체이억제Vimentin적표체,현미경하세포형태야유불규칙적사형전변위상대규칙적란원형.면역조직화학결과현시하류라서경PA-MSHA치료후간암조직E-cadherin적표체현저증강,이Snail급Slug적전록칙수도현저억제(P＜0.05).결론 PA-MSHA가이통과식별결합간암세포표면적감로당현저억제간암EMT적발생.
Objective To investigate the effects of pseudomonasaeruginosa mannose sensitive hamemagglutination vaccine (PA-MSHA) on the epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma (HCC).Methods MHCC97L and HepG2 cells were cultured and treated with different dose of PA-MSHA.After that,morphological changes of cells were observed under a light microscope.Immunofluorescence and Western blot analysis were used to evaluate the expression of EMT related genes and transcription factors.Immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) analysis were used to evaluate the expression of E-cadherin,Snail and Slug in the tumor tissue after the mice were treated with PA-MSHA.Results Western blot and immunofluorescence analysis reveled that the expression of E-cadherin was significantly elevated while expression of Vimentin was significantly inhibited by PA-MSHA,in a dose dependent manner.Moreover,the expression of fibronectin,nuclear factor-kappa B (NF-κB),Snail and Slug were also inhibited by PA-MSHA,but theses effects of PA-MSHA were blocked by mannose.After PA-MSHA treatment,HCC cells have altered from irregular shape to epithelial cobblestone appearance.In vivo study,PA-MSHA also significantly increased the expression of E-cadherin and inhibited the expression of Snail mRNA and Slug mRNA.Conclusion PA-MSHA can significantly inhibit EMT of HCC in a mannose-mediated manner.