摘要
目的探讨RNA干扰乏氧诱导因子1α(HIF-1α)对Eca109食管癌细胞放疗敏感性的影响。方法构建干扰HIF-1α质粒pSilencer 2.1 HIF-1α,采用实时荧光定量Real-time PCR、Western Blot方法验证干扰效果,通过MTT、流式细胞仪和克隆形成试验观察RNA干扰HIF-1α对Eca109食管癌细胞的放疗敏感性的影响。结果 RNA干扰可抑制Eca109细胞中HIF-1α和VEGF mRNA转录和蛋白的表达;HIF-1α基因沉默后细胞凋亡增加,且肿瘤细胞对放疗的敏感性增加。结论靶向HIF-1α的shRNA能有效抑制Eca109食管癌细胞的HIF-1α和VEGF基因表达,促进细胞凋亡,增加肿瘤细胞的放疗敏感性。
Objective To investigate the radiosensitization of silencing HIF-1α by RNA interference in human esophageal carcinoma cell lines Eeal09. Methods Vectors containing short hairpin RNA（shRNA） targeting HIF-1α gene was constructed and transfeeted into human esophageal carcinoma cell lines Eeal09, and the Real-time PCR and Western Blot were used to compare the efficiency of gene silencing. After irradiation the proliferation of cells were measured by methyhhiazolyl tetrazolium（MTT）, the cell apoptosis was detected by flow cytometry and the radiosensitivity of cells was evaluated by clone formation assay. Results After transfected with HIF-1α shRNA, the Eca109 cells mRNA and protein expression of HIF-1α and VEGF genes were in- hibited obviously. The proliferation rate of Eea1O9/shRNAs-HIF-1α group was obviously slower than control groups, and the apoptosis rate of the difference was statistically significant. The shRNA HIF-1α could also en- hance the radiosensitivity in esophageal cancer Ecal09 cells with clone formation assay. Conclusions Transfee- tion of shRNA targeting at HIF-1α gene can efficiently inhibit the expression of HIF-1α and VEGF in Ecal09 cells, which can suppress the abilities of proliferation, improve apoptosis and increase the radiosensitivity of hu- man esophageal carcinoma cell lines.
出处
《中国肿瘤外科杂志》
CAS
2012年第6期345-348,共4页
Chinese Journal of Surgical Oncology
