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[目的]讨论c-Met在食管鳞癌中的表达;构建针对人c-met基因的RNA干扰(RNAi)慢病毒载体,观察其对高侵袭性食管鳞癌细胞c-met基因的沉默效应以及对细胞生物学行为的影响。[方法]培养食管鳞癌CE81T-4细胞株,构建c-met基因的RNAi慢病毒载体,用荧光定量RT-PCR与Western blotting检测其对c-met的干扰效率,用裸鼠移植瘤模型评估RNA干扰后细胞成瘤能力的影响,用裸鼠皮下食管肿瘤生长曲线、肿瘤组织的苏木精-伊红染色及Western blotting验证c-met表达情况。[结果]成功构建c-met基因的RNAi慢病毒载体,qRT-PCR与Western blotting实验显示其对食管鳞癌CE81T-4细胞c-met mRNA及蛋白的表达的抑制均较明显,裸鼠移植瘤实验显示RNA干扰后细胞的成瘤能力降低,裸鼠皮下食管肿瘤生长曲线、肿瘤组织的苏木精-伊红染色、Western blotting实验显示裸鼠移植瘤体内c-met的表达抑制较明显。[结论]c-metRNAi慢病毒载体能够抑制食管鳞癌CE81T-4细胞的侵袭及成瘤能力。
[Objective] To investigate the expression of c-Met in esophageal squamous cell carcinoma (ESCC) and to construct RNA interference (RNAi) lentiviral vector targeting human c-met gene and to observe its silencing effect on c-met gene in highly invasive esophageal squamous cell carcinoma Impact on cell biology behavior. [Method] The esophageal squamous cell carcinoma CE81T-4 cell line was cultured and RNAi lentiviral vector of c-met gene was constructed. The interference efficiency of c-met was detected by fluorescence quantitative RT-PCR and Western blotting. The effect of RNA interference on the ability of tumorigenesis was examined. The growth curve of subcutaneous esophageal cancer in nude mice and hematoxylin-eosin staining and Western blotting were used to verify the expression of c-met. [Results] The RNAi lentiviral vector with c-met gene was successfully constructed. The results of qRT-PCR and Western blotting showed that the expression of c-met mRNA and protein in esophageal squamous cell carcinoma CE81T-4 cells were significantly inhibited. Experiments show that RNAi can reduce the ability of tumorigenesis of tumor cells. The growth curve of subcutaneous esophageal tumor in nude mice and hematoxylin-eosin staining of tumor tissue and Western blotting showed that the expression of c-met in nude mice transplanted tumor was inhibited obviously. [Conclusion] c-metRNAi lentiviral vector can inhibit the invasion and tumorigenicity of esophageal squamous cell carcinoma CE81T-4 cells.