摘要
目的:观察MACS免疫磁珠法分选CD44+CD24-/low乳腺癌干细胞活性,并检测其与多药耐药的关系。方法:运用MACS免疫磁珠法从多药耐药乳腺癌细胞株MCF-7/ADR中分选CD44+CD24-/low乳腺癌干细胞,流式细胞术测定分选前后CD44+CD24-/low细胞比例,微球体培养法检测分选细胞自我更新能力,流式检测CD44+CD24-/low细胞表面P-糖蛋白(P-gp)表达水平,Real-time PCR检测多药耐药相关基因MDR1表达水平。结果:MACS免疫磁珠法分选后,CD44+CD24-/low细胞比例为93.85%,其成球能力明显强于non-CD44+CD24-/low细胞亚群。MCF-7/ADR细胞株和CD44+CD24-/low乳腺癌干细胞P-gp表达强度分别为101 177.10±2 171.86和114 906.70±2 560.19,P<0.05。CD44+CD24-/low乳腺癌干细胞MDR1基因表达水平为MCF-7/ADR细胞株的(1.07±0.02)倍,P<0.05。结论:经MACS免疫磁珠法分选所得CD44+CD24-/low细胞亚群有更强的自我更新能力,高表达P-gp蛋白和MDR1基因可能是引起乳腺癌多药耐药的重要原因。
OBJECTIVE： To investigate the property of CD44＋CD24-/low breast cancer stem cells （BCSCs） sorted by MACS immunomagnetic beads and explore the relationship between BCSCs and muhidrug resistance （MDR）. METH- ODS： CD44＋CD24-/low breast cancer stem cells were sorted by MACS immunomagnetic beads. Flow cytometry was used to test the proportion of CD44＋CD24-/low wcells and explore the expression of P-gp. Self-renewal capacity was measured by microspheres culture assay. The expression level of muhidrug resistance gene MDR1 was detected by Real Time PCR. RESULTS： After sorting the proportion of CD44＋CD24-/low cells was 93.85%. CD44＋CD24-/low cells showed a higher self renewal capacity compared to the non-CD44＋CD24-/low cells. The expression of P-gp in MCF-7/ADR and CD44＋CD24-/low cells was 101 177.10±2 171.86 and 114 906.70±2 560.19 respectively,P〈0.05. The expression of MDR1 in CD44＋CD24-/low cells was （1.07±0.02） folds than MCF-7/ADR（P〈0.05）. CONCLUSION： CD44＋CD24-/low ceils sor- ted by MACS immunomagnetic beads have stronger self-renewal capacity and higher expression level of P-gp and MDR1, which may be an important cause of breast cancer muhidrug resistance.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2013年第12期891-893,903,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(81173601
30972932)
关键词
乳腺肿瘤
肿瘤干细胞
多药耐药
免疫磁珠分选
breast neoplasms
neoplastic stem cells
multidrug resistance
immunomagnetic separation
