CAJ | 학술논문

目的探讨前列腺癌细胞系LNCaP中分选出的干/祖(S/P)细胞上皮细胞-间充质细胞转换(EMT)的特点.方法采用流式细胞术从前列腺癌细胞系LNCaP中分选出S/P细胞,采用Western blot杂交和免疫荧光法检测上皮钙黏素(E Cadherin)、神经钙黏素(N Cadherin)、波形蛋白(Vimentin)、Snail等分子标志物的表达.采用琼脂糖凝胶克隆形成实验检测S/P细胞成瘤能力,细胞迁移实验检测细胞的迁移能力.结果与非S/P细胞相比,分选出的S/P细胞EMT标志物N Cadherin、Vimentin、Snail表达明显增强,E Cadherin的表达明显下降.细胞培养3周后非S/P细胞和S/P细胞可以在软琼脂凝胶中克隆性生长,阳性克隆分别为(18.34±1.21)个和(82.27±7.54)个,差异有统计学意义(t=8.617,P=0.001).细胞迁移48 h后,非S/P细胞迁移细胞为(25.33±5.13)个,S/P细胞为(74.33±7.64)个,差异有统计学意义(t=7.953,P=0.001).结论前列腺癌细胞系LNCaP中分选出的S/P细胞具有EMT的特点,成瘤性和迁移能力明显增强,这种现象可能促进肿瘤的侵袭和转移.
목적 탐토전렬선암세포계LNCaP중분선출적간/조(S/P)세포상피세포-간충질세포전환(EMT)적특점.방법 채용류식세포술종전렬선암세포계LNCaP중분선출S/P세포,채용Western blot잡교화면역형광법검측상피개점소(E Cadherin)、신경개점소(N Cadherin)、파형단백(Vimentin)、Snail등분자표지물적표체.채용경지당응효극륭형성실험검측S/P세포성류능력,세포천이실험검측세포적천이능력.결과 여비S/P세포상비,분선출적S/P세포EMT표지물N Cadherin、Vimentin、Snail표체명현증강,E Cadherin적표체명현하강.세포배양3주후비S/P세포화S/P세포가이재연경지응효중극륭성생장,양성극륭분별위(18.34±1.21)개화(82.27±7.54)개,차이유통계학의의(t=8.617,P=0.001).세포천이48 h후,비S/P세포천이세포위(25.33±5.13)개,S/P세포위(74.33±7.64)개,차이유통계학의의(t=7.953,P=0.001).결론 전렬선암세포계LNCaP중분선출적S/P세포구유EMT적특점,성류성화천이능력명현증강,저충현상가능촉진종류적침습화전이.
Objective To implore the characteristics of epithelial-mesenchymal transition(EMT) in human prostate cancer stem progenitor (S/P) cells isolated from LNCaP cell lines.Methods The S/P cells were obtained through florescence-activated cell sorting (FACS).Western blot and immunofluorescence assay were used to detect the S/P cells' EMT markers expression,such as E Cadherin,N Cadherin,Vimentin and Snail.Soft agar assay was used to detect the tumorigenesis ability of S/P cells.Cell migration assay was used to detect the migration ability of S/P cells.Results Compared with non S/P cells,the expressions of EMT markers,such as N Cadherin,Vimentin and Snail,were increased in S/P cells,while the expressions of epithelial marker and E Cadherin were decreased in S/P cells.After cultured for three weeks,S/P cells and non S/P cells both clonally grew.The colony numbers were (18.34±1.21) and (82.27±7.54),respectively (t =8.617,P =0.001).After cultured for 48 hours,the migration cells number was (25.33±5.13) in non S/P cells and (74.33±7.64) in S/P cells (t =7.953,P =O.001).Conclusions Human prostate cancer S/P cells isolated from LNCaP cell line have some characteristics of EMT,such as stronger tumorigenesis and migration ability,which could promote tumor invasion and metastasis.