Liu Miao, Zhou Pan
Department of Paediatrics, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.E-mail:
Hubei Medical Devices Quality Supervision and Test Institute, Wuhan 430075, Hubei Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Feb;31(1):38-44. doi: 10.19746/j.cnki.issn.1009-2137.2023.01.006.
To investigate the influence and mechanism of atorvastatin on glycolysis of adriamycin resistant acute promyelocytic leukemia (APL) cell line HL-60/ADM.
HL-60/ADM cells in logarithmic growth phase were treated with different concentrations of atorvastatin, then the cell proliferation activity was measured by CCK-8 assay, the apoptosis was detected by flow cytometry, the glycolytic activity was checked by glucose consumption test, and the protein expressions of PTEN, p-mTOR, PKM2, HK2, P-gp and MRP1 were detected by Western blot. After transfection of PTEN-siRNA into HL-60/ADM cells, the effects of low expression of PTEN on atorvastatin regulating the behaviors of apoptosis and glycolytic metabolism in HL-60/ADM cells were further detected.
CCK-8 results showed that atorvastatin could inhibit the proliferation of HL-60/ADM cells in a concentration-dependent and time-dependent manner (=0.872, =0.936), and the proliferation activity was inhibited most significantly when treated with 10 μmol/L atorvastatin for 24 h, which was decreased to (32.3±2.18)%. Flow cytometry results showed that atorvastatin induced the apoptosis of HL-60/ADM cells in a concentration-dependent manner (=0.796), and the apoptosis was induced most notably when treated with 10 μmol/L atorvastatin for 24 h, which reached to (48.78±2.95)%. The results of glucose consumption test showed that atorvastatin significantly inhibited the glycolytic activity of HL-60/ADM cells in a concentration-dependent and time-dependent manner (=0.915, =0.748), and this inhibition was most strikingly when treated with 10 μmol/L atorvastatin for 24 h, reducing the relative glucose consumption to (46.53±1.71)%. Western blot indicated that the expressions of p-mTOR, PKM2, HK2, P-gp and MRP1 protein were decreased in a concentration-dependent manner (=0.737, =0.695, =0.829, =0.781, =0.632), while the expression of PTEN protein was increased in a concentration-dependent manner (=0.531), when treated with different concentrations of atorvastatin for 24 h. After PTEN-siRNA transfected into HL-60/ADM cells, it showed that low expression of PTEN had weakened the promoting effect of atorvastatin on apoptosis and inhibitory effect on glycolysis and multidrug resistance.
Atorvastatin can inhibit the proliferation, glycolysis, and induce apoptosis of HL-60/ADM cells. It may be related to the mechanism of increasing the expression of PTEN, inhibiting mTOR activation, and decreasing the expressions of PKM2 and HK2, thus reverse drug resistance.
探讨阿托伐他汀对阿霉素耐药急性早幼粒细胞白血病（APL）细胞系HL-60/ADM糖酵解的影响及其机制。
对数生长期的HL-60/ADM细胞用不同浓度的阿托伐他汀处理，然后用CCK-8法检测细胞增殖活性，流式细胞术检测细胞凋亡，葡萄糖消耗试验检测糖酵解活性，蛋白质免疫印迹法检测PTEN、p-mTOR、PKM2、HK2、P-gp和MRP1的蛋白表达。将PTEN-siRNA转染至HL-60/ADM细胞后，进一步检测PTEN低表达对阿托伐他汀调节HL-60/ADM细胞凋亡和糖酵解代谢行为的影响。
CCK-8结果显示，阿托伐他汀能以浓度和时间依赖性方式抑制HL-60/ADM细胞的增殖（r = 0.872，r = 0.936），10 μmol/L阿托伐他汀处理24 h时增殖活性抑制最显著，降至（32.3±2.18）%。流式细胞术结果显示，阿托伐他汀以浓度依赖性方式诱导HL-60/ADM细胞凋亡（r = 0.796），10 μmol/L阿托伐他汀处理24 h时凋亡诱导最明显，达到（48.78±2.95）%。葡萄糖消耗试验结果显示，阿托伐他汀以浓度和时间依赖性方式显著抑制HL-60/ADM细胞的糖酵解活性（r = 0.915，r = 0.748），10 μmol/L阿托伐他汀处理24 h时这种抑制最明显，相对葡萄糖消耗量降至（46.53±1.71）%。蛋白质免疫印迹法表明，用不同浓度阿托伐他汀处理24 h后，p-mTOR、PKM2、HK2、P-gp和MRP1蛋白表达呈浓度依赖性降低（r = 0.737，r = 0.695，r = 0.829，r = 0.781，r = 0.632），而PTEN蛋白表达呈浓度依赖性增加（r = 0.531）。将PTEN-siRNA转染至HL-60/ADM细胞后，结果显示PTEN低表达减弱了阿托伐他汀对凋亡的促进作用以及对糖酵解和多药耐药的抑制作用。
阿托伐他汀可抑制HL-60/ADM细胞的增殖和糖酵解，并诱导其凋亡。其机制可能与增加PTEN表达、抑制mTOR激活以及降低PKM2和HK2表达从而逆转耐药有关。
