摘要
目的探讨小干扰RNA沉默激素非依赖前列腺癌DU145细胞白介素8基因对肿瘤干细胞生物学特性及耐药性的影响。方法用不同剂量的小干扰RNA沉默激素非依赖前列腺癌DU145细胞IL-8基因,应用流式仪检测ALDH阳性细胞比例的变化。采用悬浮细胞成球和克隆形成实验分别观察肿瘤干细胞自我更新和体外成瘤能力的变化。Annexin V-PE/7-AAD细胞凋亡双染试剂盒检测细胞凋亡率的变化,Western-blot法分析Akt蛋白的变化。结果 DU145细胞IL-8基因的沉默能降低肿瘤干细胞的比例,显著降低肿瘤干细胞悬浮细胞成球率和克隆形成率。多西他赛能显著抑制IL-8基因沉默DU145细胞的活性。Western-blot显示Akt的磷酸化被显著抑制。结论 IL-8基因的沉默能显著抑制激素非依赖前列腺癌干细胞的自我更新和体外成瘤能力,增加该群细胞对多西他赛的化疗敏感性,同时显著抑制了Akt的磷酸化,这可能是IL-8基因沉默抑制该群细胞干细胞特性的作用机制之一。
Objective To explore the effect of IL-8 silencing via small interfering RNA on androgen-independent prostate cancer stem cells and drug sensitive. Methods DU145 cells transfected with different concentration of IL-8 siRNA were collected. The percentage of ALDH ＋ cells were detected by flow cytometry. The sphere formation rate and cloning efficiency were determined by sphere formation assay and clonogenic assay. The viability of cells was detected by CCK-8 assay. The phosphorylation of Akt was evaluated by Western blot. Results The percentage of ALDH ＋ cells were declined under different concentration of IL-8 siRNA in DU145 cells. Sphere formation rate and cloning efficiency of prostate cancer stem cells were decreased by siRNA.The viability of IL-8 silencing DU145 cells was significantly suppressed compared with control groups and the Akt phosphorylation was activated. Conclusion IL-8 siRNA can eliminate androgen-independent prostate cancer stem cells and suppresses the selfrenew and clonogenic capacity of prostate cancer stem cells. IL-8 silencing sensitizes prostate cancer stem cells to docetaxel and inactivates the Akt pathway.
出处
《实用癌症杂志》
2016年第2期191-194,共4页
The Practical Journal of Cancer
