摘要
目的:探讨卡莫司汀(BCNU)对人脑肿瘤干细胞在细胞增殖及细胞周期方面的影响。方法:取初发的室管膜瘤手术切除标本1例,取材后制备为单细胞,分别接种于无血清培养基获得肿瘤干细胞,接种于含血清培养基获得肿瘤细胞。不同浓度的BCNU作用于细胞72h后,MTT法检测细胞在体外对BCNU的敏感性,并用终浓度为0.01μg/mL的BCNU作用细胞不同的时间段,通过流式细胞仪检测BCNU对细胞周期的影响。结果:(1)MTT显示,BCNU能抑制肿瘤干细胞及肿瘤细胞的生长,其对肿瘤干细胞(IC50=0.157μg/mL)的抑制略低于对肿瘤细胞(IC50=0.072μg/mL)(P<0.05)。(2)细胞周期方面,与阴性对照比较,两类细胞均表现为随药物作用时间的延长,S期细胞比例增高,并伴有凋亡细胞的增多,但肿瘤干细胞的改变(24h)出现晚于肿瘤细胞(6h)。结论:BCNU对体外培养的脑肿瘤干细胞生长有明显的抑制作用,作用机制可能为通过干扰S期DNA的合成起作用;与脑肿瘤细胞相比,脑肿瘤干细胞在MTT及细胞周期的检测中,均表现出对BCNU有一定的耐药性。
Objective To investigate the effects of BCNU on the proliferation and cell cycle of stem cells from human brain tumor. Methods Single cell suspensions generating from a specimen of primary ependymocytoma were cultured separately in serum-free and serum-supplemented medium to harvest BTSCs and BTCs. The cells were then treated with BCNU at different concentrations for 72 h and their sensitivity to BCNU in vitro was measured by MTT assay. They were also treated with BCNU at 0.01 μg/mL at different time points, then cell cycle was examined by flow cytometry. Results BCNU could inhibit the growth of both BTSCs and BTCs, with a slight lower level in BTSCs than in BTCs （0.157 μg/mL vs 0.072μg/mL for IC50, P〈0.05）. As compared with the negative controls, the proportion of the cells in S phase was increased with the prolonged treatment of BCNU, so was the amount of the apoptotic cells. However, the changes of BTSCs were behind that of BTCs （24 h vs6 h）. Conclusions BCNU has a marked inhibitory effect on the growth＇ of BTSCs cultured in vitro, whose mechanism may be involved in interfering DNA synthesis in the S phase. BTSCs reveal some resistance to BCNU as compared with BTCs.
出处
《实用医学杂志》
CAS
2008年第17期2933-2936,共4页
The Journal of Practical Medicine
关键词
脑肿瘤
肿瘤干细胞
细胞周期
细胞增殖
Brain neoplasms Tumor stem cell Cell cycle Cell proliferation
