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

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。

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

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.