盐霉素对低分化鼻咽癌干细胞放疗敏感性的影响

Effects of salinomycin on radiation sensitivity of low differentiation nasopharyngeal carcinoma stem cells

目的探讨盐霉素对低分化鼻咽癌干细胞放疗敏感性的影响。方法将鼻咽癌干细胞悬液放入标有1~24的孔板中培养,其中1~12为对照组,13~24为实验组,其中向对照组加入200μl的PBS缓冲液,向实验组加入200μl终浓度为2μmol/L的盐霉素,适宜环境中培养12 h后,两组细胞均给予6 Gy单次照射,放疗结束后,接种至培养皿中常规培养2 w,GIEMSA染色,计算细胞存活分数(SF),采用RT-PCR及Western印迹法检测选取培养24 h后人鼻咽癌干细胞中Rad51、Ku70、Ku80基因及蛋白表达水平。结果实验组培养皿中细胞SF明显低于对照组(P<0.05)。实验组培养皿鼻咽癌干细胞Rad51、Ku70和Ku80 mRNA表达水平低于对照组(P<0.05)。实验组培养皿鼻咽癌干细胞Rad51、Ku70和Ku80蛋白表达水平明显低于对照组(P<0.05)。结论盐霉素能够明显抑制人低分化鼻咽癌干细胞中Rad51、Ku70和Ku80表达,降低细胞存活分数,提高放疗敏感性和临床疗效。

Objective To investigate the effects of salinomycin on radiation sensitivity of low differentiation nasopharyngeal carcinoma stem cells. Methods Nasopharyngeal carcinoma stem cell suspension were put into orifice plate marked with 1 ~ 24,plated with 1 ~ 12 were selected as the control group,with 13 ~ 24 were selected as the experimental group. 200 μl PBS were added into control group,200 μl salinomycin( concentration was 2 μmol / L) were added into experimental group. Cells were cultured into suitable environment for 12 h,and were given 6 Gy single radiation therapy. After radiation therapy,cells were cultured into petri dish for 2 weeks. Cell survival fraction( SF)was calculated by GIEMSA staining,Rad51,Ku70,Ku80 gene and protein expression were detected by RT-PCR and Western blot after cultured 24 h. Results SF in experimental group was lower than that of control group( P < 0. 05). Expression of Rad51,Ku70,Ku80 mRNA in experimental group were lower than those of control group( P < 0. 05). Expression of Rad51,Ku70,Ku80 protein in experimental group were lower than those of control group( P < 0. 05). Conclusions Salinomycin could significantly inhibit the expressions of Rad51,Ku70 and Ku80 in low differentiation nasopharyngeal carcinoma cells,reduce the SF,improve radiation sensitivity and clinical curative effect.