免疫磁珠分选的口腔鳞状细胞癌干细胞诱导分化及成瘤活性研究  

Differentiation and tumor-forming activity of oral squamous cell carcinoma stem cells selected by immunomagnetic beads

作　　者：廖显翔 房芳 羊良慧 陈国生 王代友[1] 彭剑波[1] 刘诗奇 麦华明[1] LIAO Xianxiang;FANG Fang;YANG Lianghui;CHEN Guosheng;WANG Daiyou;PENG Jianbo;LIU Shiqi;MAI Huaming(Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Guangxi Medical University;Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction;Guangxi Universities and Colleges Key Laboratory of Oral and Maxillofacial Surgery Disease Treatment;Guangxi Clinical Research Center for Craniofacial Deformity, Nanning 530021, China)

机构地区：[1]广西医科大学附属口腔医院口腔颌面外科广西口腔颌面修复与重建研究重点实验室广西高校颌面外科疾病诊治研究重点实验室广西颅颌面畸形临床医学研究中心

出　　处：《中国癌症防治杂志》2019年第2期131-137,共7页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：国家自然科学基金项目(81460413)

摘　　要：目的探讨口腔鳞状细胞癌干细胞的诱导分化及成瘤活性。方法采用免疫磁珠分选法从6例原代培养的口腔鳞状细胞癌细胞中分选出CD133^+、CD44^+细胞,以流式细胞术检测其纯度及细胞周期,并绘制CD133^+细胞生长曲线。将分选出的细胞经诱导向成骨肪细胞及成脂肪细胞分化,将原代鳞癌细胞、分选所得的CD44^+细胞及CD133^+细胞进行体内成瘤实验。结果经免疫磁珠分选法成功获得CD44^+细胞、CD133^+细胞,其纯度经检测均在94%以上,且主要处于G0/G1期。CD133+细胞在接种后第2天进入对数期,细胞倍增时间为3.22 d。经诱导后CD44^+、CD133^+细胞成功分化为成骨细胞及成脂肪细胞。皮下注射后第3天可触及新生肿物,在3种细胞成瘤实验中CD133^+细胞组瘤体体积最大,HE染色显示符合低分化口腔鳞状细胞癌病理表现。结论免疫磁珠分选法可高效获得高纯度的口腔鳞状细胞癌干细胞。CD44^+、CD133^+细胞均具有较强的多向分化能力,CD133^+细胞成瘤性较强。Objective To investigate the differentiation and tumor-forming aitivity of oral squamous cell carcinoma stem cells. Methods The CD133^+ and CD44^+ cells were isolated by immunomagnetic beads from oral squamous carcinoma cells,and the purity of sorted cells and cell cycle were identified by flow cytometry.The CD133^+ cell growth curve was determined. The selected cells were induced to differentiate into osteoblasts and adipocytes.In vivo tumorigenesis experiments were carried out using primary squamous cells and CD44^+,CD133^+ cells. Results The purity of CD44^+ cells and CD133+ cells that were isolated by immuno-magnetic beads were more than 94% detected by flow cytometry. Most of their cell cycle were in G0/G1 phase.CD133^+ cells entered log phase on the 2nd day after inoculation,and the cell doubling time was 3.22 d.After induction,CD44+ and CD133+ cells were successfully differentiated into osteoblasts and adipocytes. On the 3rd day after subcutaneous injection,new tumors could be touched, the tumor volume in the CD133+ cell group was the greatest in three cells,and HE staining showed that it was consistent with the pathological manifestations of oral squamous cell carcinoma. Conclusion High-purity oral squamous cell carcinoma stem cells can be successfully sorted by immunomagnetic beads method.CD44+ and CD133+ cells have great multidirectional differentiation,but the tumorigenicity of CD133+ is greater.

关 键 词：口腔鳞状细胞癌 磁珠分选 干细胞 CD44 CD133 诱导分化 

分 类 号：R1739.8[医药卫生—妇幼卫生保健]