摘要
目的研究P1WIL2在人膀胱癌肿瘤于细胞样细胞（CSLC）中的表达和意义，探讨PIWIL2化靶向肿瘤干细胞治疗中的作用。方法用无血清悬浮培养法从人膀胱移行细胞癌细胞株BIU-87中分离扶得悬浮细胞球，流式细胞仪检测细胞表面分子标志CDl33和CD44的表达，免疫磁珠分选系统分离CDl33＋C1）44＋细胞；分刖采用Transwell小室侵袭实验、膜联蛋白／碘化丙锭双标流式细胞术、半定量逆转录聚合随链反应（RT—PCR）检测CSLC的体外侵袭能力、凋亡率和PIWIL2 mRNA的表达。结果BIU-87细胞可在无血清培养攮小形成可悬浮生K、稳定传代的肿瘤干细胞样细胞球。与贴壁的BIU-87细胞相比，CSLC高表达CDl33和CD44，具仃更强的体外侵袭力、较低的凋亡率，P1WIl2mRNA在CSLC的表达也显著高于BIU-87细胞。结论采用尤血清悬浮培养法从人膀胱移行细胞癌细胞株中分离获得的CSLC，具有自我更新平和多分化的能力，高懂表达PIWIL2。
Abstract cells （ CSLC ） from human bladder transitional cell carcinoma and study the role of PIWIL2 in the targeting therapy of cancer stem cells. Methods The method of suspension culture in specific serum free medium was employed to isolate and obtain cancer stem - like cells spheres from human bladder transitional cell carcinoma line BIU - 87. The expressions of cell surface markers CD133 and CD44 were detected by flow cytometry. CD133 ＋ CD44 ＋ cells were separated with magnetic activated cell sorting system. Transwell invasive method, Annexin - V/PI double - labeled flow cytometry technique and semi -quantitative reverse transcription polymerase chain reaction （RT- PCR） were performed to detect the invasive ability, apoptosis rate and P1WI12 mRNA expression level of CSLC. Results CSLC derived from BIU - 87 cells were obtained and formed floating cell spheres under the suspension culture melhod with serum - free medium, and could propagate stably, which highly expressed CD133 and CD44 of cancer stem cell markers, with stronger invasive ability and lower apoptosis rate compared with BIU - 87 cells. And PIWIL2 mRNA expression level in CSLC was significantly higher than that in BIU -87 cells. Conclusions CSLC can be isolated and enriched under the suspension cuhure method with serum - free medium, which possess the potentials of self - renewal and differentiation. Therefore, PIWIL2 is highly expressed in CSLC.
出处
《国际泌尿系统杂志》
2011年第6期722-726,共5页
International Journal of Urology and Nephrology
基金
国家自然科学基金青年基金（30801138）
广东省自然科学基金面上项目（S2011010004230）
广东省医学科研基金面上项目（A2011632）
韶关市医药卫生科研汁划面上项目（Y11019）
关键词
癌
侈行性细胞
膀胱肿瘤
肿瘤千细胞
Carcinoma, Transitional Cell
Bladder Neoplasms
Tumor Stem Cells
