摘要
目的从胃癌细胞株中分离并培养侧群(SP)细胞,探讨其耐药性及致瘤性。方法应用流式细胞术从胃癌BGC-823细胞株中分选SP细胞,MTT法观察SP细胞和非SP细胞的增殖情况,流式细胞术检测不同浓度氟尿嘧啶(5-FU)对BGC-823细胞增殖抑制作用以及SP细胞比例、细胞周期情况;通过在NOD/SCID鼠体内种植不同数量的SP细胞和非SP细胞,比较两者的致瘤能力。结果流式细胞仪检测胃癌BGC-823细胞中存在0.9%～2.1%的SP细胞,MTT法显示SP细胞有较强的增殖活性。2.5、5、10和25μg/ml 5-FU作用于BGC-823细胞(2.0×106个)36h后,细胞数分别为2.6×106、2.3×106、2.0×106和1.8×106,SP细胞比例依次为12.9%、9.9%、4.0%和2.1%,SP细胞处于G0/G1期。SP细胞和非SP细胞分别以1×104细胞数接种NOD/SCID鼠,SP细胞形成瘤体的平均瘤重为0.577g,而非SP细胞为0.150g,且SP细胞成瘤时间较非SP细胞短且瘤体数量多、质量高。结论人胃癌细胞系BGC-823中的SP细胞具有较强的增殖活性和类似干细胞的耐药性,针对SP细胞可以为抗肿瘤治疗提供新的靶点。
Objective To observe the drug resistance and tumorigenic ability of side population cells（SP cells） which were sorted from BGC-823 cell lines.Methods SP cells were collected from BGC-823 cells by flow cytometry（FCM）.The cell proliferation of SP cells and non-SP cells were measured by methabenzthiazuron（MTT） assay.The FCM was employed to investigate the inhibitory effect of different concentrations of 5-fluorouracil（5-FU） on BGC-823 cells and measure the proportion and cell cycle of SP cells.The tumorigenic ability of the SP cells was assessed by in vivo transplantation into non-obese diabetic/severe combined immuno-deficiency（NOD/SCID） mice.Results There was 0.9%-2.1% SP cells in BGC-823 cells.It showed that SP cells had a higher proliferative ativity than non-SP cells by MTT assay.The cell numbers of BGC-823 treated by 2.5,5,10,25μg/ml 5-FU for 36h were 2.6×106,2.3×106,2.0×106,1.8×106;and the proportions of SP cells were 12.9%,9.9%,4.0%,2.1%,respectively.At the same time,it demonstrated that sorted SP cells has the tougher viability than non-SP cells when suffering with drug treatment.It showed that SP cells were in G0/G1 phase by FCM.The average weights of tumors formed by SP cells（1×104） and non-SP cells（1×104） were 0.577g and 0.150g,respectively.SP cells possessed higher tumorigenicity than non-SP cells,such as tumor growth time and tumor weight.Conclusion SP cells from human gastric cancer cell line BGC-823 possessed a higher proliferative activity and stem cell-like drug-resistence character.The research of SP cells could provide new targets for anti-tumor therapy.
出处
《临床肿瘤学杂志》
CAS
2012年第8期673-677,共5页
Chinese Clinical Oncology
基金
国家自然科学基金资助项目(30672771)
关键词
胃癌
肿瘤干细胞
氟尿嘧啶
致瘤能力
Gastric carcinoma
Cancer stem cells
5-fluorouracil
Tumorigenic ability
