摘要
目的探讨人IL-18基因联合二萜生物碱对舌鳞癌细胞Tscca的增殖抑制与促凋亡作用。方法构建pEGFPN3-IL-18质粒,转染舌鳞癌细胞Tscca。荧光显微镜下观察质粒的转染效率;MTT检测pEGFPN3-IL-18联合二萜生物碱对舌鳞癌细胞Tscca生长的影响;流式细胞术检测细胞凋亡情况;蛋白质免疫印迹法检测细胞信号调节激酶Akt/p-Akt的蛋白水平。结果转染pEGFPN3-IL-18的舌鳞癌细胞Tscca较未转染细胞凋亡数增加,差异有统计学意义(P<0.05);0.2、0.4、0.6mg/mL二萜生物碱培养舌鳞癌细胞,随浓度增加,细胞凋亡数增多,差异有统计学意义(P<0.05);IL-18与二萜生物碱联合对Tscca细胞作用48h后,较单独使用二萜生物碱的增殖抑制作用明显,且呈浓度依赖性,差异有统计学意义(P<0.05);IL-18联合二萜生物碱还可剂量依赖性地降低p-Akt的蛋白水平。结论pEGFPN3-IL-18联合二萜生物碱对舌鳞癌细胞Tscca有协同抑制作用。
Objective To investigate the effects of human IL-18 gene combined with diterpenoid alkaloids in inhibiting the proliferation and inducing the apoptosis of tongue squamous carcinoma cells Tscca.Methods We constructed recombinant plasmid pEGFPN3-IL-18 and tranfected it into tongue squamous carcinoma cells Tscca.The transduction efficiency of the target cells was detected by fluorescent microscopy,cytotoxic effect of IL-18 gene with diterpenoid alkaloids on Tscca was detected by MTT assay,and apoptosis was detected by flow cytometry.Western blot was employed to examine the expression level of cellular signal-regulated kinase Akt/p-Akt.ResultsThe tongue squamous cells Tscca which transfected pEGFPN3-IL-18 had significantly increased apoptosis compared with non-transfected cells（P〈0.05）.Tongue carcinoma squamous cells cultured with diterpenoid alkaloids at the concentrations of 0.2,0.4 and 0.6 mg/mL had significantly increased apoptosis in a dose-dependent manner（P〈0.05）.Human IL-18 gene combined with diterpenoid alkaloids for 48 hours inhibited significantly Tscca in a concentration-dependent manner compared with diterpenoid alkaloids alone（P〈0.05）.The two in combination could also decrease the protein level of p-Akt dose-dependently.Conclusion The combination of pEGFPN3-IL-18 and diterpenoid alkaloids has a synergistic effect in inhibiting the growth of tongue squamous carcinoma cells Tscca.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2017年第2期215-220,共6页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
辽宁医学院校长基金-奥鸿博泽研究生科技创新基金(No.AH2015007)~~
