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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 observe the activity of CD44 + CD24- / low breast cancer stem cell sorting by MACS immunomagnetic beads and to detect its relationship with multidrug resistance. Methods: CD44 + CD24- / low breast cancer stem cells were sorted out by multi-drug resistant breast cancer cell line MCF-7 / ADR using MACS immunomagnetic beads method. CD44 + CD24- / low cells were detected by flow cytometry The expression of P-glycoprotein (P-gp) on CD44 + CD24- / low cells was detected by flow cytometry. The expression of multidrug resistance-related gene MDR1 was detected by Real-time PCR Level. Results: The percentage of CD44 + CD24- / low cells after MACS immunomagnetic beads sorting was 93.85%, which was significantly stronger than that of non-CD44 + CD24- / low cells. The P-gp expression intensity of MCF-7 / ADR cell line and CD44 + CD24- / low breast cancer stem cells were 101 177.10 ± 2 171.86 and 114 906.70 ± 2 560.19 respectively, P <0.05. The MDR1 gene expression level of CD44 + CD24- / low breast cancer stem cells was (1.07 ± 0.02) times of MCF-7 / ADR cell line, P <0.05. CONCLUSION: The CD44 + CD24- / low cell subpopulations isolated by MACS immunomagnetic beads method have stronger self-renewal ability. High expression of P-gp protein and MDR1 gene may be the important reason of multidrug resistance in breast cancer.