姜黄素对食管癌EC-109细胞增殖抑制的研究

目的检测姜黄素对食管癌EC-109细胞的增殖抑制作用,并从细胞周期角度探讨其分子机制。方法以体外培养的食管癌EC-109细胞株作为研究对象,分为实验组和对照组,实验组给予不同浓度姜黄素,对照组给予等剂量生理盐水。20、40、80μmol/L姜黄素分别作用于食管癌Ec-109细胞,采用MTT法检测细胞增殖抑制率,应用流式细胞仪检测细胞周期各时相分布情况及计算细胞增殖指数,Western-blot检测细胞周期蛋白D1、E(cyclin D1、cyclin E)表达情况。结果 20、40、80μmol/L姜黄素分别作用食管癌EC-109细胞12、24、48 h后,与对照组比较,细胞增殖抑制率随着药物作用时间延长及剂量加大而增高,有统计学意义(F=71.40,P<0.00)。相同时间点不同浓度组间比较及相同浓度不同时间组间比较均有统计学意义(P<0.01),说明姜黄素呈现时间-剂量依赖性抑制细胞增殖。上述浓度姜黄素干预24 h后,G0/G1期细胞比例逐步增大,有统计学意义(P=6.27,P<0.05),不同浓度组间比较均有统计学意义(P<0.05),说明姜黄素阻滞EC-109细胞于G0/G1期呈剂量依赖关系。同样条件下,Western-blot检测到cyclin D1、cyclin E蛋白条带逐渐变窄,说明姜黄素能抑制周期蛋白表达。结论姜黄素通过下调食管癌EC-109细胞表达cyclin D1、cyclin E,使细胞阻滞于G/G期,起到抑制细胞增殖的作用。

The Study of the Proliferation Inhibition of Curcumin on EC-109 Cells

Objective To observe the effect of curcumin inhibit human esophageal carcinoma EC-109 cells proliferation,and try to explain it by way of cell cycle change.Methods Moderately differentiated human esophageal carcinoma EC-109 cells were treated with 20,40 and 80 μmol/L curcumin in vitro for 12,24 and 48 hour respectively,the control group was treated with normal saline.The proliferation-inhibiting capability was analyzed with MTT assay.Cell cycle were detected by flow cytometry.The expression of cyclin D1 and cyclin E was detected by Western-blot.Results After EC-109 cells were treated with 0,20,40 and 80μmol/L curcumin for 12,24 and 48 hour respectively,the ratio of proliferation-inhibiting was increased gradually when time went on or concentration of curcumin was added gradually,the change was statistically significant(P<0.01).It showed that curcumin effectively inhibited the proliferation of EC-109 cells in a concentration-time dependent way.EC-109 cells of G0/G1 phase increased gradually in a concentration-dependent way when EC-109 cells were treated with different concentration curcumin for 24 hour(P<0.05).The proliferation index was lessened in a concentration-dependent way too(P<0.05),which was(47.55±2.36)%,(40.75±4.23)%,(31.26±3.67)% and(22.78±2.02)% respectively when EC-109 cells were treated for one day with 0,20,40 and 80μmol/L curcumin respectively,the expression of cyclin D1 and cyclin E was also presented a tendency of down-regulation.Conclusion Curcumin can inhibited human esophageal carcinoma EC-109 cells line proliferation obviously in vitro,which is through inducing EC-109 cells stagnation at G0/G1 phase by down-regulated the expression of cyclin D1 and cyclin E.