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中国癌症防治杂志 ›› 2019, Vol. 11 ›› Issue (2): 131-137.doi: 10.3969/j.issn.1674-5671.2019.02.09

• 基础研究 • 上一篇    下一篇

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

  

  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

  • 出版日期:2019-04-25 发布日期:2019-05-29
  • 通讯作者: 麦华明。 E-mail:dddmai@126.com
  • 基金资助:
    国家自然科学基金项目(81460413)

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

  • Online:2019-04-25 Published:2019-05-29
  • Contact: MAI Huaming. E-mail:dddmai@126.com

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。

关键词: 口腔鳞状细胞癌, 磁珠分选, 干细胞, CD44, CD133, 诱导分化

Abstract: 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.

Key words: Oral squamous cell carcinoma, Magnetic bead sorting, Stem cells, CD44, CD133, Differentiation

中图分类号: 

  • R739.8