CAJ | 학술논문

目的：从人舌鳞癌组织中分离培养肿瘤干细胞细胞（CSC），鉴定其生物学特性，并研究PIWIL4在CSC中的表达和意义。方法收集9例不同临床分期舌鳞癌患者组织标本，通过酶消化和原代培养相结合等方法处理，采用无血清悬浮培养法分离培养获得含CSC的悬浮细胞球，流式细胞仪检测细胞表面分子标志CD133和CD44的表达，免疫磁珠分选系统分离CD133+CD44+细胞；采用半定量逆转录聚合酶链反应检测PIWIL4 mRNA在CSC中的表达；裸鼠皮下接种CSC，观察其成瘤能力。结果成功的从人舌鳞癌组织分离培养获得可悬浮生长、稳定传代的CSC，CSC高表达CD133和CD44，裸鼠皮下接种1×104、1×105个CSC均可全部成瘤，PIWIL4 mRNA在CSC的表达也显著高于正常舌组织细胞。结论采用无血清悬浮培养法成功从人舌组织中分离获得CSC，具有肿瘤干细胞特性，能高度表达PIWIL4，为靶向肿瘤干细胞的治疗提供了实验依据。
목적：종인설린암조직중분리배양종류간세포세포（CSC），감정기생물학특성，병연구PIWIL4재CSC중적표체화의의。방법수집9례불동림상분기설린암환자조직표본，통과매소화화원대배양상결합등방법처리，채용무혈청현부배양법분리배양획득함CSC적현부세포구，류식세포의검측세포표면분자표지CD133화CD44적표체，면역자주분선계통분리CD133+CD44+세포；채용반정량역전록취합매련반응검측PIWIL4 mRNA재CSC중적표체；라서피하접충CSC，관찰기성류능력。결과성공적종인설린암조직분리배양획득가현부생장、은정전대적CSC，CSC고표체CD133화CD44，라서피하접충1×104、1×105개CSC균가전부성류，PIWIL4 mRNA재CSC적표체야현저고우정상설조직세포。결론채용무혈청현부배양법성공종인설조직중분리획득CSC，구유종류간세포특성，능고도표체PIWIL4，위파향종류간세포적치료제공료실험의거。
Objective To isolate and cultivate cancer stem-like cells (CSC) from human squamous cell carcinoma of tongue and identify their biological characteristics, investigate the expression and signiifcance of PIWIL4 in CSC. Methods Tumor samples were obtained from 9 patients of carcinoma of the tongue at stages T3-T4. The method of suspension culture in speciifc serum free medium was employed to isolate and obtain CSC spheres by using enzymatic digestion combined with primary culture. 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. Semi-quantitative reverse transcription polymerase chain reactions were performed to detect the PIWIL4 mRNA expression in CSC. The tumor formation ability was evaluated by subcutaneous injection of CSC into nude mice. Results CSC were obtained from human squamous cell carcinoma of tongue and formed lfoating cell spheres under the suspension culture method with serum-free medium, and could propagate stably, which highly expressed CD133 and CD44 of CSC markers. And PIWIL4 mRNA expression level in CSC was signiifcantly higher than that in normal tongue tissues. Tumor formated in all mice after subcutaneous injection of 1×104 and 1×105 CSC and exhibited a high degree of similarity with the primary tumor in carcinoma of the tongue. Conclusion CSC can be isolated under the suspension culture method with serum-free medium, which possess the potentials of CSC and highly expressed PIWIL4.