苦马豆素对骨肉瘤类肿瘤干细胞N-糖链表达的影响

The influence of swainsonine bring in the expression of N-glycan of osteosarcoma tumor stem cells

目的:检测苦马豆素对骨肉瘤类肿瘤干细胞(CSCs)N-糖链表达的影响。方法:通过无血清培养法培养小鼠骨肉瘤细胞株LM8,经磁珠细胞分选技术选得CD133及CD44双阳性LM8细胞,使用不同剂量的N糖基化抑制剂苦马豆素处理分选得到的骨肉瘤CSCs,通过凝集素结合实验(L-PHA)检测细胞表面β1,6分支N-糖链的表达水平。结果:通过无血清培养初步分离后再进行磁珠分选得到的细胞中CD133和CD44双阳性细胞比例为(96.5±1.2)%,骨肉瘤CSCs与L-PHA结合率达(90.3±2.1)%,高于LM8细胞的结合率(54.3±3.1)%(P<0.05)。用1 mg/m L的苦马豆素处理骨肉瘤CSCs后,细胞表面β1,6分支N-糖链的表达阳性率为90%,用5 mg/m L的苦马豆素处理骨肉瘤CSCs后,细胞表面β1,6分支N-糖链的表达阳性率为21%。结论:N糖基化抑制剂苦马豆素能抑制骨肉瘤CSCs表面β1,6分支N-糖链的表达。

Objective Detecting the influence of swainsonine bring in the expression of N- glycan of osteosarcoma tumor stem eells. Methods LM8 cells were cultured in the culture medium （free of serum）, then selected the LM8 ceils of both CD44 and CD133 positive by magnetic activated cell sorting technique. Detecting the levels of 131, 6 branch N-glycan expression in the selected LM8 cells （treating by different doses of N-glycosylation inhibitor swainsonine） surface through the lectin binding assay （L-PHA）. Results （96.5 ± 1.2）% of LM8 cells were both CD133 and CD44 positive which selected from all the LM8 cells in cuhure medium （free of serum） by magnetic activated cell sorting technique. Lectin binding assays shown that the bound rare of osteosarcoma tumor stem cells and L-PHA was （90.3 ±2,1）%, which was higher than normal LM8 cells and L-PHA by （54.3± 3.1）% （P 〈 0.05）. The positive rare of β1, 6 branch N-glycan on the selected LM8 cells（treat with lmg/mL swainsonine） surface was 90%. The positive rare of β1, 6 branch N-glycan on the selected LM8 cells （treat with 5mg/mL swainsonine） surface was 21%. Conclusion N-glyeosylation inhibitor swainsonine plays a role in inhibiting the expression of N-glycan on cell surface of osteosarcoma tumor stem cell.