γ-synuclein在食管癌中的表达及其对食管癌Eca-109细胞侵袭力的影响

Expression of γ-synuclein in esophageal carcinoma and its effect on invasion of esophageal carcinoma Eca-109 cells

目的:研究γ-synuclein在食管癌中的表达及其对人食管癌细胞株Eca-109侵袭力的影响。方法:选取2010年1月至12月于河北医科大学第四医院食管癌患者组织标本30例,同时取10例正常食管组织作对照。采用免疫组化法观察食管癌组织中γ-synuclein的表达情况。采用脂质体法转染含γ-synuclein的重组质粒pcDNA3.0-γ-synuclein,应用RT-PCR检测转染后Eca-109细胞中γ-synuclein mRNA的表达,流式细胞术检测γ-synuclein蛋白的表达,利用Transwell实验检测转染后Eca-109细胞侵袭力的变化。结果:与正常食管组织相比,食管癌组织中γ-synuclein蛋白阳性表达率明显增强(83.3%vs40.0%,P<0.05),Ⅰ-Ⅱ期食管癌患者γ-synuclein表达率明显低于Ⅲ-Ⅳ期(22.2%vs 76.2%,P<0.05)。pcDNA3.0-γ-synu-clein质粒转染后,Eca-109细胞中γ-synuclein mRNA表达水平显著高于空质粒转染组和未转染组[(1.10±0.03)vs(0.42±0.03),(0.45±0.15),P<0.01],蛋白表达水平也明显增高[(1.05±0.061)vs(0.80±0.45),(0.79±0.46);P<0.05],穿膜细胞数明显增加[(167±2.51)vs(65±2.60),(70±2.50);P<0.01]。结论:食管癌组织高表达γ-synuclein,其表达上调能增强人食管癌Eca-109细胞的侵袭力,提示其在食管癌的转移和恶性发展中发挥重要作用。

Objective ： To detect the expression of γ-synuclein in esophageal adenocarcinoma and evaluate its effect on invasion of esophageal carcinoma Eca-109 cells. Methods： SP immunohistochemical staining was used to examine the expression of γ-synuclein in 30 esophageal carcinoma specimens and 10 normal control specimens from January to December 2010 （Fourth Hospital of Hebei Medical University）. The plasmid containing γ-synuclein （pcDNA3.0-γ-synuclein） was transfected into Eca-109 cells by lipofectamine. The expressions of γ-synuclein mRNA and protein were investigated by RT-PCR and flow cytometry, respectively. The invasion ability of transfected Eca-109 cells was dectected by Transwell assay. Results： The positive exppression rate of γ-synuclein was 83% in human esophageal carcinoma, which was significantly higher than that in normal esophageal tissues （P〈0.05）, and the positive rate of γ-synuclein in stage-Ⅱ esophageal carcinoma patients was significantly lower than that in stage Ⅲ-Ⅳ patients （22.2% vs 76.2%,P〈0.05）. By comparing transfected group in Eca-109 cells with empty plasmid group and control group, RT-PCR showed the expression level of γ-synuclein mRNA increased significantly （／[1.10±0.03／] vs ／[0.42±0.03／],／[0.45±0.15／]; P〈0.01）. Flow cytometry showed that the expression level of γ-synuclein protein increased significantly compared with the other two groups （／[1.05±0.06／] vs ／[0.80±0.45／], ／[0.79±0.46／]; P〈0.05）. Invasion through matrigel abilities of Eca-109 cells increased significantly （／[167±2.51／] vs ／[65±2.60／], ／[70±2.50／]; P〈0.01）. Conclusion： γ-synuclein is highly expressed in esophageal carcinoma tissues and can enhance the invasion of Eca-109 cells, which may play an important role in carcinogenesis progression and metastasis of esophageal carcinoma.