前列腺癌细胞系中干祖细胞的分选和鉴定  

Isolation and identification of stem/progenitor cells from human prostate cancer cell line

作　　者：马志方[1] 许召良[1] 岳亮[1] 郝振文[1] 王东文[1] 

机构地区：[1]山西医科大学第一医院泌尿外科,太原030001

出　　处：《中华细胞与干细胞杂志（电子版）》2012年第4期21-25,共5页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：山西省留学回国人员科技活动择优资助项目(晋财社[2011]172);山西省高等学校留学回国人员科研资助项目(晋外教[2011]63);山西省回国留学人员科研资助项目(2012-085)

摘　　要：目的探讨在前列腺癌细胞系LNCaP中分选和鉴定干祖(S/P)细胞的方法。方法常规培养前列腺癌细胞系LNCaP,用CD133和CD44抗体标记细胞,流式细胞术分选,条件培养液培养分选出的S/P细胞,用定量RT-PCR、Westernblot和免疫荧光染色方法检测S/P细胞CD133、α2β1整合素、雄激素受体(AR)、CK5、CK8和PSA的表达,前列腺细胞球体形成实验检测S/P细胞自我更新能力,琼脂糖凝胶克隆形成实验检测S/P细胞成瘤能力。各指标两组间差异比较采用独立t检验。结果采用流式细胞术可以分选出约占1﹪的S/P细胞,与非S/P细胞相比,S/P细胞CK5、CD133、α2β1整合素表达增高(mRNA表达比较,t=17.376,3.859,8.611;P=0.000,0.021,0.001),CK8、AR和PSA的表达明显下降(mRNA表达比较,t=3.518,11.372,3.845;P=0.029,0.000,0.018)。细胞培养14d后,非S/P细胞和S/P细胞形成前列腺球的数量分别为(4.35±0.87)个和(35.12±4.56)个,两者比较差异具有统计学意义(t=13.013,P=0.000)。细胞培养14d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,阳性克隆数分别为(18.34±1.21)个克隆和(82.27±7.54)个克隆,两者比较差异有显著意义(t=8.617,P=0.001)。结论前列腺癌LNCaP细胞系中存在占极少数的S/P细胞,荧光激活流式细胞术可成功分选出S/P细胞。Objective To isolate and identify the stern/progenitor （S/P） cells from human prostate cancer cell line LNCaE Methods LNCaP cells were marked by CD133 and CD44 antibodies. The SP cells were obtained through florescence-activated cell sorting （FACS）. Quantitative RT-PCR, Western blot and immunofluorescence assay were used to detect the SP cells＇ markers, such as CD 133 and a2131 integrin. Differentiated cell markers Androgen receptor （AR）, CKS, CK8 and PSA were detected simultaneously. Sphere formation assay was used to detect the self-renewing ability of SP cells. Soft agar assay was used to detect the tumorigenesis ability of SP cells. Results About 1% SP cells could be obtained. Compared to non SP cells, CKS,CD133 and ct2131 integrin expression was higher in SP cells （mRNA expression t = 17.376, 3.859, 8.611; P = 0.000, 0.021, 0.001）.

关 键 词：前列腺癌 干细胞 CD133 CD44 

分 类 号：R737.25[医药卫生—肿瘤]