CAJ | 학술논문

　　目的：研究Akt活性抑制剂LY294002抑制源自人小细胞肺癌NCI-H446细胞系肺癌球形成细胞即肺癌干细胞样细胞(LCSLCs)自我更新作用。方法：体外培养NCI-H446细胞系细胞。以干细胞条件培养基用超低粘附6孔细胞培养板悬浮培养富集和扩增LCSLCs。裸鼠皮下成瘤实验鉴别LCSLCs高致瘤特性。Western blot分析LCSLCs中Akt蛋白磷酸化水平。肿瘤球形成试验检测LY294002对LCSLCs自我更新的影响。结果：干细胞条件培养基悬浮培养6d，NCI-H446细胞系细胞呈三维非粘附性球体生长。 LCSLCs具有高致瘤特性。与NCI-H446细胞系细胞比较，LCSLCs信号分子Akt组成性活化。 LY294002有效降低LCSLCs中Akt磷酸化水平，并以剂量依赖方式抑制LCSLCs肺癌球形成(P＜0.05)。结论：靶向干预小细胞肺癌LCSLCs信号分子Akt组成性活化可能成为抑制肺癌干细胞特性治疗小细胞肺癌的新策略。
　　목적：연구Akt활성억제제LY294002억제원자인소세포폐암NCI-H446세포계폐암구형성세포즉폐암간세포양세포(LCSLCs)자아경신작용。방법：체외배양NCI-H446세포계세포。이간세포조건배양기용초저점부6공세포배양판현부배양부집화확증LCSLCs。라서피하성류실험감별LCSLCs고치류특성。Western blot분석LCSLCs중Akt단백린산화수평。종류구형성시험검측LY294002대LCSLCs자아경신적영향。결과：간세포조건배양기현부배양6d，NCI-H446세포계세포정삼유비점부성구체생장。 LCSLCs구유고치류특성。여NCI-H446세포계세포비교，LCSLCs신호분자Akt조성성활화。 LY294002유효강저LCSLCs중Akt린산화수평，병이제량의뢰방식억제LCSLCs폐암구형성(P＜0.05)。결론：파향간예소세포폐암LCSLCs신호분자Akt조성성활화가능성위억제폐암간세포특성치료소세포폐암적신책략。
Objective To investigate LY294002, a inhibitor specific to Akt activities, suppress the self-re-newal of lung cancer stem cell like cells (LCSLCs) from small cell lung cancer cell line NCI-H446 cell line cul-tured in vitro. Methods Human small cell lung cancer NCI-H446 cell line was cultured in vitro. Cells were plat-ed in stem cell conditioned culture system allowed for sphere forming, namely LCSLCs. In vivo tumorigenicity ex-periments were used to examine height tumorigenicity of LCSLCs. The phosphorylation level of signaling molecule Akt protein was determined using Wester bolt. Tumor sphere formation assay was used to the inhibitory effects of LY294002 on the self-renewal of LCSLCs. Results The small cell lung cancer cells were plated in stem cell con-ditioned culture medium in 6-well plates at a density of 5,000 cells/well which allowed for the formation of colonies separated from each other for 6d. LCSLCs had the height tumorigenicity in vivo in nude mouse model. The phosphorylation level of signaling molecule Akt protein in LCSLCs was heighter than those of small cell lung cancer cell line NCI-H446 cell line cultured in vitro. LY294002 significantly reduced the phosphorylation level of Akt protein in LCSLCs, and inhibited pulmosphere formation, in a concentration-dependent manner. Conclu-sion Targeting intervention of the constructively activation of signaling molecule Akt may can provide a novel strategy for small cell lung cancer therapy with activity of inhibition of LCSLCs characteristics.