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桔梗皂苷D(platycodin D,PD)对多种恶性肿瘤有显著抑制作用,对白血病细胞有明显的增殖抑制和诱导凋亡的作用,但其是否有效提高耐药细胞对伊马替尼的敏感性及其发挥功能的分子机制仍未阐明。为了研究PD与伊马替尼(imatinib,IM)联合用药对慢性粒细胞白血病耐药细胞株K562/R的作用及机制。细胞增殖实验检测桔梗皂苷D对伊马替尼增殖抑制功能的影响,采用CCK8测定PD和IM单药及联合用药对K562/R增殖的抑制作用;流式细胞术检测Annexin V-FITC/PI双标记细胞凋亡率的变化,Western blot法检测cleaved caspase-3,cleaved caspase-9,PARP,cleaved PARP,Bcr/abl,p-AKT,p-m-TOR蛋白表达。结果显示桔梗皂苷D联合伊马替尼对K562/R细胞的增殖抑制作用和细胞凋亡率比单独用药组效果明显。Western blot法检测结果显示与单药组比较,联合用药组可以明显上调cleaved caspase-3,cleaved caspase-9,cleaved PARP蛋白表达,同时下调PARP,Bcr/abl,p-AKT,p-mTOR蛋白的表达。结果表明桔梗皂苷D可以提高耐药细胞对伊马替尼的敏感性,联合用药在抑制细胞增殖、诱导凋亡、抑制Bcr/abl蛋白和激活PI3K/AKT/mTOR信号通路方面明显优于单独用药。
Platycodin D (PD) has a significant inhibitory effect on many malignant tumors, and has obvious effects on inhibiting proliferation and inducing apoptosis of leukemia cells, but whether it can effectively increase the sensitivity of the resistant cells to imatinib And the molecular mechanism of its function has not been elucidated yet. To investigate the effect and mechanism of combination therapy of PD and imatinib (IM) on K562 / R cell line resistant to chronic myeloid leukemia. Cell proliferation assay was used to detect the inhibitory effect of platycodin D on the proliferation inhibition of imatinib. The inhibitory effect of PDK and IM monotherapy on the proliferation of K562 / R cells was determined by CCK8. Flow cytometry was used to detect Annexin V-FITC / The changes of apoptosis rate were detected by Western blot. The expressions of cleaved caspase-3, cleaved caspase-9, PARP, cleaved PARP, Bcr / abl, p-AKT and pm- The results showed that the inhibitory effect of platypodin D combined with imatinib on the proliferation of K562 / R cells and the apoptosis rate were more obvious than those of the drug alone group. The results of western blot showed that compared with the single drug group, the combined treatment group could obviously upregulate the expression of cleaved caspase-3, cleaved caspase-9 and cleaved PARP, and down-regulate the expressions of PARP, Bcr / abl, p-AKT and p-mTOR expression. The results showed that platycodin D could increase the sensitivity of drug-resistant cells to imatinib. The combination therapy was significantly better than that of imatinib alone in inhibiting cell proliferation, inducing apoptosis, inhibiting Bcr / abl protein and activating PI3K / AKT / mTOR signaling pathway .