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目的:探讨益气养阴方对CD34+CD38-KG1a白血病干细胞活性的影响。方法:从KG1a细胞株中分选出CD34+CD38-KG1a干细胞,采用CCK-8法检测益气养阴方及联合注射用盐酸柔红霉素(DNR)对KG1a干细胞的抑制率;流式细胞术检测益气养阴方干预KG1a干细胞后各组细胞周期及凋亡比例;RT-PCR法检测益气养阴方干预KG1a干细胞后各组PTEN、m TOR m RNA的表达量。结果:经过免疫磁珠分选后CD34+CD38-KG1a细胞比例为(97.20±2.53)%。益气养阴方+DNR组48、72h抑制率明显增高(P<0.05),益气养阴方组抑制率未见增高。益气养阴方组和益气养阴方+DNR组G0/G1期比例明显降低,G2/M期比例明显增高(P<0.05)。益气养阴方组凋亡细胞比例增高,但死亡细胞未见增高,益气养阴方+DNR组的凋亡和死亡细胞均明显增高(P<0.05)。益气养阴方组、益气养阴方+DNR组PTEN m RNA表达增高(P<0.05),但益气养阴方+DNR组m TOR m RNA表达较其他3组均明显下降(P<0.05)。结论:益气养阴方对KG1a干细胞不具有直接抑制作用,但可促进DNR对KG1a干细胞的抑制作用,其机制可能是益气养阴方通过活化PTEN基因负调控PI3K/Akt信号通路,从而影响KG1a干细胞的细胞周期和凋亡。
Objective: To investigate the effect of Yiqi Yangyin Recipe on the activity of CD34 + CD38-KG1a leukemia stem cells. Methods: CD34 + CD38-KG1a stem cells were sorted out from KG1a cell line. The inhibition rate of KG1a stem cells was measured by CCK-8 method and Qi-qi nourishing Yin combined with daunorubicin hydrochloride (DNR) The effect of Yiqi Yangyin Prescription on the cell cycle and the percentage of apoptotic cells in KG1a stem cells were detected. The expression of PTEN and m TOR m RNA in KG1a stem cells were measured by RT-PCR. Results: The percentage of CD34 + CD38-KG1a cells after immunomagnetic bead sorting was (97.20 ± 2.53)%. The inhibition rate of Yiqi Yangyin Prescription + DNR group was significantly increased at 48 and 72h (P <0.05), while the inhibition rate of Yiqi Yangyin Prescription group was not increased. The proportion of G0 / G1 phase in Yiqi Yangyin Prescription and Yiqi Yangyin Prescription + DNR group was significantly lower, and the proportion of G2 / M phase was significantly increased (P <0.05). The proportion of apoptotic cells in Yiqi Yangyin prescription group increased, but the death cells did not increase. The apoptosis and death cells in Yiqi Yangyin prescription + DNR group were significantly increased (P <0.05). The expression of PTEN m RNA in Yiqi Yangyin prescription group and Yiqi Yangyin prescription + DNR group increased (P <0.05), but m TOR m RNA expression in Yiqi Yangyin prescription + DNR group was significantly lower than that in the other three groups (P < 0.05). Conclusion: Yiqi Yangyin Prescription has no direct inhibition on KG1a stem cells, but it can promote the inhibitory effect of DNR on KG1a stem cells. The mechanism may be that Yiqi Yangyin Prescription can negatively regulate PI3K / Akt signaling pathway by activating PTEN gene Cell Cycle and Apoptosis of KG1a Stem Cells.