摘要
目的探讨转染sh Artemis干扰质粒对人肝癌细胞BEL-7402/5FU DNA损伤的影响。方法将人肝癌细胞分为正常对照组、脂质体对照组、空质粒对照组和转染sh Artemis干扰质粒实验组(sh Artemis实验组);建立DNA损伤模型,分别用0.625、1.25、2.5、5.0和10.0μg/m L的丝裂霉素C作用BEL-7402/5FU细胞24和48 h,MTT法检测细胞活性,Western blot观察磷酸化组蛋白H_2AX(γ-H_2AX)的表达;转染Artemis干扰质粒,检测Artemis、γ-H_2AX表达;彗星实验检测DNA的损伤。结果 0.625、1.25、2.5、5.0和10.0μg/m L的丝裂霉素C作用48h后肝癌细胞活力明显下降(P<0.05);丝裂霉素C刺激细胞48 h后,γ-H_2AX的表达量随着药物浓度的增加而增加;转染sh Artemis干扰质粒后,γ-H_2AX的表达量较正常对照组增加(P<0.05);sh Artemis实验组较正常对照组拖尾DNA量增加(P<0.05)。结论丝裂霉素C能够诱导肝癌细胞损伤;转染sh Artemis干扰质粒促进丝裂霉素C诱导的人肝癌细胞BEL-7402/5FU DNA损伤。
Objective To explore the cellular effects of shArtemis on DNA damage of human hepatic carcinoma cell line BEL-7402/5FU in vitro. Methods The experiment samples were divided into control group, Lipo- fectamine2000 group, contol plasmid group, shArtemis group;Human hepatic carcinoma cell line BEL-7402/SFU were treated with 0. 625,1.25,2.5.5.0 and 10.0 μg/mL Mitomycin C for 24 and 48 huors. BEL-7402/SFU cell activity examined by MTT and observed the expression of phosphorylated histone2AX （γ-H2AX） by Western blot. After 48 h transfected shArtemis interference plasmid, Western blot detected the quantity of Artemis and γ-H2AX; Commet assy detect the exent of DNA damage. Results The MTT results showed that after 48 h treat- ment within 0. 625, 1.25, 2.5, 5.0 and 10.0 μg/mL mitomycin C, cell activity of liver cancer significantly de- creased. In addition, after 48 h of incubation with mitomycin C, the expression of γ-H2AX increased in a dose- dependent manner; The the expression of γ-H2AX increased following inhibitated Artemis. Moreover the tailingDNA of shArtemis experimental group increased than in the control group. Conclusions Mitomycin C can induce DNA damage in human hepatic carcinoma cell line BEL-7402/SFU; shArtemis prmotes DNA damage induced by mitomyein C.
出处
《基础医学与临床》
CSCD
2016年第3期353-357,共5页
Basic and Clinical Medicine
基金
贵州省社发攻关项目([2009]3066)
遵义市红花岗区科技基金([2009]18)
贵州省教育厅特色重点实验室建设项目([2014]212)
