miR126抑制食管鳞癌血管内皮生长因子表达的实验研究

  目的  探讨miR126与血管内皮生长因子在食管鳞癌中的表达及关系, 以及其对食管鳞癌细胞系的影响。  方法  随机选取2011年3月至2011年4月间手术治疗的食管鳞癌患者手术标本4例, , 取癌组织及癌旁正常黏膜。提取肿瘤及正常癌旁组织microRNA, 逆转录后用实时定量PCR的方法检测miR126的表达情况; 提取标本中的mRNA和蛋白用实时定量PCR和Westernblot的方法检测VEGF的表达; 培养TE-1细胞系并转染miR126模拟体, 检测miR126对TE-1细胞增殖和VEGF表达的影响。  结果  与正常组织相比, miR126在癌组织的表达明显降低; 在RNA和蛋白水平, VEGF水平均明显升高; 转染miR126模拟体的细胞BrdU掺入量较转染阴性对照的细胞明显增多, VEGF在RNA和蛋白水平上均明显降低。  结论  在食管鳞癌组织中miR126水平下调, VEGF表达增高; miR126可抑制TE-1细胞的增殖, 其抑制细胞增殖的作用可能是通过下调VEGF的表达而实现的。 

Experimental study on the miR126 inhibition of vascular endothelial growth factor expression in esophageal squamous cell carcinoma

  Objective  This study aimed to explore miR126 expression and its relationship with vascular endothelial growth factor(VEGF) in esophageal squamous cell carcinoma, as well as the effect of miR126 expression on an esophageal carcinoma cell line.  Methods  Four patients with esophageal squamous cell carcinoma were randomly selected in our hospital from 2011.3 to 2011.4.Tumor and normal tumor-adjacent tissue specimens were collected from the patients, and miRNA was extracted.The expression of miR126 was detected by real-time quantitative PCR.Furthermore, total mRNA and protein were extracted to detect VEGF expression by real-time quantitative PCR and Western blot analysis.miR126-like mRNA was then transfected to the TE-1 cell line to determine its effect on the proliferation of TE-1 cells and VEGF expression.  Results  miR126 expression was significantly downregulated in esophageal carcinoma tissues compared with normal esophageal tissues.By contrast, the expression of VEGF RNA and protein significantly increased in esophageal carcinoma tissues compared with normal esophageal tissues.The incorporation of BrdU in cells transfected with miR126-like mRNA significantly increased compared with untransfected cells, whereas the levels of VEGF RNA and protein significantly decreased in transfected cells.  Conclusion  miR126 expression was downregulated and VEGF was upregulated in esophageal squamous cell carcinoma.Therefore, miR126 may downregulate the expression of VEGF and inhibit the proliferation of TE-1 cells.