免疫磁珠分选与顺铂富集喉癌Hep-2细胞系中肿瘤干细胞的比较

Comparation of magnetic activated cell sorting and chemotherapy agent DDP in enriching cancer stem cell of laryngeal carcinoma cell line Hep-2

目的比较免疫磁珠分选（MACS）及化疗药物顺铂（DDP）筛选富集喉癌Hep-2细胞系肿瘤干细胞的效果。方法以CD133MACS、DDP作用两种方法来富集喉癌Hep-2细胞系中的肿瘤干细胞，并以流式细胞术（FCM）检测处理后CD133^＋细胞的百分率。同时观察细胞形态学的改变，判断两种方法分选后细胞对后续实验的影响。结果经FCM检测，MACS分选喉癌Hep-2，CD133^＋细胞得率为64．33％，不同浓度DDP作用于喉癌Hep-2细胞48h，FCM检测CD133^＋细胞有不同的得率，其中质量浓度为4μg／ml时，所得CD133^＋≥细胞百分率最高，为50．7％。MACS与DDP各组比较差异均有统计学意义（P〈0．01）；两种方法处理后MACS组细胞的存活状态要好于DDP组。结论MACS分选纯度较高，对细胞损伤小，适合后续培养；但分选前耗时长，每次分选仅能用于一种Marker。DDP筛选简单易行，符合临床肿瘤干细胞（CSC）抵抗化疗的模式。但阳性细胞得率与细胞毒性成正比。MACS和DDP筛选肿瘤干细胞各有其优势及适用范围，实验中可根据不同目的来确定所用筛选方法。

Objective To compare effect of chemotherapy agent DDP to MACS in sorting cancer stem cells （CSC） of laryngeal carcinoma cell line Hep-2. Methods CD133 magnetic beads were applied to sort Hep-2 cells. Different dosages of DDP were used to treat Hep-2 cells for 48 hours. Enrichment rate of CD133^＋ cells by MACS and after DDP treatment was detected by Flow Cytometer （FCM）. Morphologic change was observed under inverse-phase microscope. Results FCM showed that the sorting rate of CD133^＋ cells through MACS was 64.33 %, while after DDP treatment for 48 hours, the rate of CD133^＋ cells was enriched significantly in each dosage of DDP, with the maximal rate was 50.7 %, in the dosage of 4 μg/ml. There was a significantly difference between MACS and each of DDP group （P 〈0.01）. Cells treated with DDP were abnormal in morphology. Conclusion MACS and DDP sorting has respective advantages in enriching CSC in Hep-2 cell lines.