柯里拉京对胶质瘤U251细胞及其干细胞增殖活性影响初探

Influence of corilagin on the proliferation in U251 glioma cells and glioma stem cells

目的从胶质瘤U251细胞中分离肿瘤干细胞并探讨柯里拉京对胶质瘤U251细胞及其干细胞增殖活性和细胞周期的影响。方法使用免疫磁珠法联合无血清培养法分离肿瘤干细胞,50μg/ml柯里拉京分别干预U251细胞及U251干细胞12 h、24 h,CCK-8检测干预后细胞存活率改变,流式细胞术检测细胞周期变化,倒置显微镜观察干预前后细胞的形态学变化。结果胶质瘤U251细胞中含有少量CD133+细胞,在无血清培养基中呈悬浮、球形生长,具有典型的干细胞特性,在有血清培养基中能够自我分化为胶质瘤细胞;柯里拉京可明显抑制胶质瘤U251细胞及其干细胞的增殖,且对胶质瘤干细胞的抑制作用强于胶质瘤细胞(P<0.05);柯里拉京可使胶质瘤U251细胞停留在G2/M期,使U251干细胞停留在S期(P<0.05)。结论胶质瘤U251细胞中含有少量肿瘤干细胞;柯里拉京可抑制胶质瘤细胞及其干细胞的增殖,影响细胞周期,但其对U251细胞及其干细胞的细胞周期影响不一,机制可能存在差异。

Objective To isolate the CD133＋ cancer stem cells fiom U251 cells and discuss the influence of corilagin on U251 cells and cancer stem cells. Methods Glioma stem cells were isolated fi＇om U251 cells by immunomagnetic beads technique and serum-flee culture method. The corilagin of 50 i^g/ml was used to interfere the U251 cells and U251 stem cells for 12 h and 24 h. CCK-8 was used to detect the survival rate of the cells after intervention. Cell cycle distribution was analyzed by flow cytometry and the morphological changes of the cells were observed before and after intervention under microscope. Results There were a small number of CD 133＋ cells in U251 cells. This kind of cells maintained suspension and sphere-like growth status in serum-free medium in vitro and showed characteristics of cancer stem cells, and could differentiate themselves to gliomas cells in serum medium. Corilagin inhibited the cell proliferation of U251 cells and their stem cells and the inhibition role was stronger in U251 stem cells （P 〈0.05）. Corilagin made the U251 cells stay in G2/M phase and the U251 stem cells in S phase （P 〈 0.05）. Conclusions There is a small number of tumor stem cells in U251 ceils. Corilagin can inhibit the cell proliferation and influence the cell cycle both in U251 cells and U251 stem cells, but there are different mechanisms between them.