^125I粒子对胆管细胞癌HCCC-9810细胞凋亡及Livin基因表达的影响

Effect of 125I seeds brachytherapy on Livin expression and apoptosis of HCCC-9810 cholangiocarcinoma cells

目的探讨^125I粒子对胆管细胞癌HCCC-9810细胞凋亡及Livin基因表达的影响。方法实验分为空白对照组、空壳组、^125I粒子组(根据临床常用放射性活度分为:0.5 mCiGy ^125I粒子组、0.7 mCiGy ^125I粒子组、0.9 m CiGy ^125I粒子组)。体外培养HCCC-9810细胞,实验组经相应活度^125I粒子干预48 h后,通过免疫组化法、RT-PCR法分别检测各组细胞Livin蛋白及mRNA的表达,同时应用流式细胞术(FCM)检测各组细胞的凋亡率变化。结果与空白对照组比较,空壳组Livin蛋白和mRNA的表达及细胞凋亡率差异无统计学意义(P>0.05);与空白对照组及空壳组比较,^125I粒子组(0.5、0.7和0.9 mCiGy)中胆管细胞癌HCCC-9810细胞的凋亡率及Livin基因表达水平同时增加,差异有统计学意义(P<0.05)。0.5 mCiGy ^125I粒子组中HCCC-9810细胞的Livin蛋白和mRNA的表达水平最低而细胞凋亡率最大,0.7 mCiGy ^125I粒子组HCCC-9810细胞Livin蛋白和mRNA表达水平最高,但细胞凋亡率却最低,0.9 mCiGy ^125I粒子组中HCCC-9810细胞的Livin蛋白和mRNA的表达水平及细胞凋亡率介于两者之间。结论不同剂量的^125I粒子诱导胆管细胞癌HCCC-9810细胞凋亡的能力存在差异,其作用机制与其调节细胞内Livin基因表达水平高低密切有关。Livin作为凋亡抑制基因,其表达水平的高低可能与胆管癌细胞放射抵抗性有一定的相关性。

Objective To observe the effects of 125I seeds brachytherapy on apoptosis of HCCC-9810 cholangiocarcinoma cells and Livin expression.Methods The study included 5 groups:blank control group,vacant shell group,and 125I seeds group(0.5 mCiGy,0.7 mCiGy and 0.9 mCiGy group).HCCC-9810 cholangiocarcinoma cells were cultured in vitro.RT-PCR was used to detect the expression of Livin mRNA in HCCC-9810 cholangiocarcinoma cells 48 h after treatment with 125I seeds.The expression of Livin protein in the cells was indicated using Immunohistochemistry and Western Blot.Flow cytometry was performed to assess apoptosis by using annexin V-fluorescein isothiocyanate(Annexin V-FITC)/PI double staining.Results Compared with blank control group and vacant shell group,cell apoptosis rates of the three experimental groups were increased(P<0.05),also,the expression of Livin was significantly enhanced(P<0.05).Among the three experimental groups,the lowest expression of Livin along with the highest cell apoptosis rate were detected in 0.5 mCiGy 125I seeds group;but the expression of Livin in 0.7 mCiGy 125I seeds group was higher than that of another two groups,accompanying with the lowest cell apoptosis rate.However,there was no significant difference in Livin expression and apoptosis rate between blank control group and vacant shell group.Conclusion There is significant difference in ability to induce cell apoptosis among different doses of 125I,which may be related to the ability to elevate Livin expression in HCCC-9810 cholangiocarcinoma cells.Radio-resistance may be related to the level of Livin(an apoptosis suppressor gene)gene expression in cholangiocarcinoma cells.