摘要
目的 观察重组人血管内皮抑素（rhES）对食管鳞癌KYSE-150细胞的放射敏感性的影响及其机制的初步探讨.方法 将食管鳞癌KYSE-150细胞分为空白对照组、rhES组、X射线照射组和rhES联合X射线照射组.细胞克隆形成法测定细胞存活分数；单击多靶模型拟合细胞存活曲线并计算放射增敏比（SER）；流式细胞术检测rhES联合X射线照射对细胞周期及凋亡的影响；反转录聚合酶链反应（RT-PCR）检测Cyclin B1、Cyclin D1、Bcl-2、Bax mRNA表达水平,Western blot检测HIF-1α、VEGF、VEGFR蛋白的表达水平.结果 KYSE-150细胞的Do、Dq、SF2值随着rhES浓度的增加逐渐减小,当达到Do照射剂量时,100和200 μg/ml rhES的放射增敏比分别为1.14、1.27；rhES联合X射线照射组与X射线照射组比较,细胞凋亡率增加、凋亡相关基因bax的表达增加、细胞VEGF蛋白及HIF-1α蛋白的表达水平降低（t=7.97、3.02、117.55、7.22,P＜0.05）.结论 rhES对体外食管癌细胞具有明显的辐射增敏作用,其机制与其抑制细胞HIF-1 α、VEGF蛋白表达以及对细胞凋亡相关基因bax的表达调控、诱导细胞凋亡密切相关.
Objective To study the effect of recombinant human endostatin （rhES）on the radiosensitivity of esophageal squamous cells KYSE-150 and its preliminary mechanism.Methods Cells were divided into four groups：control group without treatment,rhES group treated with recombinant human endostatin,radiation alone group exposed with X-rays,and combination group exposed with X-rays plus endostatin.Colony formation assay was used to measure cell survival fraction.A single-hit multi-target model was used to fit cell survival curve and calculate the sensitive enhancement ratio （SER）.Influence of rhES combined with X-ray radiation on cell cycle and apoptosis was measured by flow cytometry.Expressions of Cyclin B1,Cyclin D1,Bcl-2 and Bax mRNAs were determined by RT-PCR.Protein expressions of HIF-1α,VEGF,and VEGFR were determined by Western blot.Results D0,Dq and SF2 value of KYSE-150 cells decreased along with the concentration of rhES.At D0dose,the SER for 100 and 200 μg/ml rhES was 1.14 and 1.27,respectively.Compared with the radiation alone group,the apoptosis rate and bax expression increased,while the expressions of VEGF and HIF-1α decreased in the combination group （t =7.97,3.02,117.55,7.22,P 〈 0.05）.Conclusions rhES has radiosensitive effect on esophageal carcinoma cells KYSE-150 in vitro by inhibiting the expressions of HIF-1α and VEGF,regulating bax expression,and inducing apoptosis.
出处
《中华放射医学与防护杂志》
CAS
CSCD
北大核心
2013年第4期346-350,共5页
Chinese Journal of Radiological Medicine and Protection
基金
江苏省卫生厅指导性科研项目（Z201220）
常州市卫生局重大项目（ZD201105）
常州市科技支撑社会发展项目（CE20125021）
关键词
重组人血管内皮抑素
食管癌细胞
辐射敏感性
Recombinant human endostatin
Esophageal carcinoma cells
Radiosensitivity
