Hao Y, Ji J, Liu C, Zhang N, Gong Y
Key Laboratory of Cancer Research and Clinical Laboratory Diagnosis, Bengbu Medical College, Bengbu 233030, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2022 Nov 20;42(11):1739-1746. doi: 10.12122/j.issn.1673-4254.2022.11.20.
To investigate the effects of matrine combined with LY294002 on proliferation, apoptosis and cell cycle of human myeloid leukemia K562 cells and explore the underlying mechanism.
The effects of different concentrations of matrine alone and in combination with LY294002 on the proliferation of K562 cells were examined with CCK-8 assay. The changes in morphology of K562 cells were observed following treatment for 48 h with 0.4 g/L matrine and 10 μmol/L Y294002, either alone or in combination, and cell apoptosis was detected using flow cytometry with annexin V-FITC/PI double labeling; the changes in cell cycle was detected with PI labeling. Western blotting was performed to examine the effect of matrine combined with LY294002 on expressions of p-mTOR, p-PI3K, Akt, p-Akt, cyclinD1, Bcl-2 and caspase-9 in the cells.
Treatment with different concentrations of matrine, both alone and in combination with LY294002, inhibited the proliferation of K562 cells in a time- and concentration-dependent manner. Compared with matrine treatment alone, the combined treatment caused more obvious morphological changes of the cells, significantly increased cell apoptosis ( < 0.01), and induced cell cycle arrest in G/G ( < 0.01). Western blotting showed that the protein expression levels of p-mTOR, cyclinD1, p-PI3K, p-Akt and Bcl-2 in K562 cells increased while the expression level of caspase-9 decreased significantly after the combined treatment ( < 0.01).
Matrine combined with LY294002 produces a synergistic inhibitory effect on K562 cells possibly by down-regulating the p-Akt expression in PI3K/Akt signaling pathway, reducing the expressions of p-mTOR, cyclinD1 and Bcl-2, and increasing the expression of caspase-9.
探讨苦参碱联合LY294002对人髓系白血病K562细胞增殖、凋亡及细胞周期的影响，并探讨其潜在机制。
采用CCK-8法检测不同浓度苦参碱单独及联合LY294002对K562细胞增殖的影响。用0.4 g/L苦参碱和10 μmol/L LY294002单独及联合处理K562细胞48 h后，观察细胞形态变化，采用Annexin V-FITC/PI双染流式细胞术检测细胞凋亡；用PI标记检测细胞周期变化。采用蛋白质免疫印迹法检测苦参碱联合LY294002对细胞中p-mTOR、p-PI3K、Akt、p-Akt、cyclinD1、Bcl-2和caspase-9表达的影响。
不同浓度苦参碱单独及联合LY294002处理均能抑制K562细胞增殖，且呈时间和浓度依赖性。与单独使用苦参碱处理相比，联合处理引起细胞形态变化更明显，细胞凋亡显著增加（P<0.01），并诱导细胞周期阻滞于G0/G1期（P<0.01）。蛋白质免疫印迹法显示，联合处理后K562细胞中p-mTOR、cyclinD1、p-PI3K、p-Akt和Bcl-2蛋白表达水平升高，而caspase-9表达水平显著降低（P<0.01）。
苦参碱联合LY294002可能通过下调PI3K/Akt信号通路中p-Akt的表达，降低p-mTOR、cyclinD1和Bcl-2的表达，增加caspase-9的表达，对K562细胞产生协同抑制作用。
