人骨肉瘤细胞MG-63侧群细胞的分离及耐药性研究  

Isolation of side population cells from the MG-63 osteosarcoma cell line and characterization of their chemotherapeutic resistance

作　　者：陈晓明[1] 苏节[2] 李刚[2] 钟艳平[2] 

机构地区：[1]广西医科大学第一附属医院脊柱骨病外科,南宁530021 [2]广西医科大学医学科学实验中心

出　　处：《中国癌症防治杂志》2015年第3期196-200,共5页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：广西自然科学基金资助项目(No 2012GXNSFAA053134);广西区域性高发肿瘤防治研究重点实验室项目(GK2014-ZZ17)

摘　　要：目的分选人骨肉瘤细胞MG-63中的侧群(side population,SP)细胞,并探讨SP细胞对化疗药物耐药的机制。方法以流式细胞术仪检测、分选经DNA染料Hoechst 33342染色后MG-63中的SP细胞,采用CCK-8法检测顺铂对SP细胞与非侧群(non-SP)细胞的抑制率,并计算半数抑制浓度(IC50)。荧光定量PCR法检测SP细胞和non-SP细胞中MRP、XRCC1、MGMT、ABCG2 m RNA的表达水平。结果 MG-63细胞在细胞密度为9×105个/ml、Hoechst 33342终质量浓度为5μg/ml时,MG-63 SP细胞的比例最稳定,为(1.45±0.23)%,且细胞保持良好的活性。顺铂对SP细胞的IC50为11.608μg/ml,对non-SP细胞的IC50为6.876μg/ml,两者比较差异有统计学意义(P<0.05)。MRP、XRCC1、MGMT和ABCG2 m RNA在SP细胞中的表达分别是non-SP细胞的1.58倍、1.61倍、2.36倍和3.14倍(P<0.05)。结论人骨肉瘤MG-63细胞中存在SP细胞亚群,与non-SP细胞相比,SP细胞对顺铂的耐药性更为明显,可能与其耐药基因MRP、XRCC1、MGMT、ABCG2m RNA的高水平表达有关。Objective To isolate side population（SP）cells and non-SP cells from the MG-63 human osteosarcoma cell line and investigate the mechanism behind SP cell resistance to chemotherapy. Methods MG-63 SP cells were isolated by flow cytometry using the blue fluorescent dye Hoechst 33342.Resistance of SP and non-SP cells to Cisplatin（DDP）were analyzed using the Cell Counting Kit-8,and 50% inhibitory concentration （IC50） was calculated for both subpopulations. Expression of MRP,XRCC1,MGMT and ABCG2 transcripts was assayed by RT-PCR in SP and non-SP cells. Results SP cells accounted for （1.45±0.23）%of MG-63 cells at a density of 9× 105/ml after staining with 5μg/ml Hoechst 33342. DDP showed an IC50 of 11.608 g/mL in SP cells,significantly higher than the value in non-SP cells（P〈0.05）. Transcript levels were significantly higher in SP cells than in non-SP cells for MRP（1.58-fold higher）,XRCC1（1.61-fold）,MGMT（2.36-fold）and ABCG2（3.14-fold）（P〈0.05）. Conclusion MG-63 human osteosarcoma cells contain SP cells,which show higher resistance to DDP than non-SP cells. Up-regulation of MRP,XRCC1,MGMT and ABCG2 in SP cells may play a role in this drug resistance.

关 键 词：骨肿瘤 侧群细胞 肿瘤干细胞 耐药性 

分 类 号：R738.1[医药卫生—肿瘤]