摘要
[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 evaluate the expression of c-Met in esophageal squamous cell carcinoma by constructing lentiviral vectors for RNA interference（RNA）of c-met gene and detecting its silencing effect on the biological behaviors of human esophageal squamous cell carcinoma CE81T-4cells.[Methods]Human esophageal squamous cell carcinoma CE81T-4cells were cultured.Lentiviral for RNA of c-met gene was constructed and the silencing effect was detected by RT-PCR and Western blotting.In vivo Tumorigenicity assay was performed to analyze in vivo proliferation of esophageal squamous cell carcinoma CE81T-4cells in a xenograft model.[Results]Lentiviral RNAi vectors targeting c-met gene were successfully constructed,and they efficiently inhibited the expression of c-met at mRNA and protein levels.Transfection of c-met lentivial RNAi vectors inhibited the invasion and tumorigenicity of CE81T-4cells.[Conclusion]Lentivirusmediated c-met RNAi efficiently inhibits invasion and tumorigenicity of CE81T-4cells.
出处
《中国中西医结合消化杂志》
CAS
2016年第11期847-850,共4页
Chinese Journal of Integrated Traditional and Western Medicine on Digestion
基金
新疆自治区重大科技专项计划(No:201430123-1)
