作　　者： ; ;

机构地区： 中南大学湘雅医院

出　　处： 《上海交通大学学报（医学版）》 2019年第9期991-997,991,共7页

摘　　要： 目的·探讨3-羟基-3-甲基-辅酶A合成酶1(3-hydroxymethyl-3-methylglutaryl-CoAsynthase1,HMGCS1)对于急性髓细胞性白血病(acute myelocytic leukemia,AML)HL-60细胞株药物敏感性的作用机制。方法·培养HL-60细胞,分别通过感染阴性对照慢病毒和HMGCS1慢病毒构建阴性对照组和HMGCS1过表达组细胞株,设置未作处理的HL-60细胞为空白对照组。采用实时荧光定量PCR(real-time quantitative PCR,qPCR)检测3组细胞中HMGCS1mRNA含量,并验证HMGCS1过表达组细胞株是否构建成功。应用Westernblotting检测HMGCS1对磷脂酰肌醇3激酶(phosphatidylinositol 3 kinase,PI3K)/蛋白激酶B(protein kinase B,PKB,又称AKT)信号通路中AKT及磷酸化AKT表达水平的影响。采用CCK8法检测HMGCS1及PI3K/AKT信号通路抑制剂LY29400对HL-60细胞活力的影响。应用qPCR和Western blotting检测LY29400对HMGCS1表达水平的影响。结果·与阴性对照组相比,HMGCS1过表达组细胞中HMGCS1 mRNA水平显著增加(P=0.000)。与空白对照组及阴性对照组相比,HMGCS1过表达组细胞中的磷酸化AKT的表达水平明显升高,而AKT的水平则无明显差异。与空白对照组及阴性对照组相比,HMGCS1可降低阿霉素对细胞活力的影响(P=0.003,P=0.006),而LY294002则可抑制由HMGCS1产生的作用(P=0.000)。在阴性对照组及过表达组细胞中,予以LY294002干预可降低HMGCS1 mRNA(均P=0.000)和蛋白的表达水平。结论·HMGCS1可以降低HL-60细胞对化学治疗药物阿霉素的敏感性,而PI3K/AKT信号通路抑制剂LY294002则可恢复其敏感性。 Objective · To explore the mechanism of 3-hydroxymethyl-3-methylglutaryl-CoA synthase 1 (HMGCS1) on drug sensitivity of acute myelocytic leukemia (AML) HL-60 cells. Methods · HL-60 cells were cultured. The negative control group and the HMGCS1 overexpressed group were constructed by infecting the negative control lentivirus and HMGCS1 lentivirus, and the untreated HL-60 cells were set as the blank control group. Real-time quantitative PCR (qPCR) was used to detect the expression of HMGCS1 mRNA in the 3 groups, and to verify whether the cell lines of the HMGCS1 overexpressed group were successfully constructed. The effect of HMGCS1 on the expression of AKT and phosphorylated AKT (p-AKT) in phosphatidylinositol 3 kinase (PI3K)/ protein kinase B (PKB / AKT) signaling pathway was detected by Western blotting. CCK8 method was used to detect the effects of HMGCS1 and PI3K/AKT signaling pathway inhibitor LY29400 on the activity of HL-60 cells. The effect of LY29400 on HMGCS1 expression was detected by qPCR and Western blotting. Results · Compared with the negative control group, the HMGCS1 mRNA expression was increased significantly in the HMGCS1 overexpressed group (P=0.000). Compared with the blank control group and the negative control group, the p-AKT protein level in the HMGCS1 overexpression group was significantly increased, while the AKT expression of the 3 groups was not significantly different. CCK8 method showed that compared with the blank control group and the negative control group, HMGCS1 could reduce the effect of adriamycin on cell viability in the HMGCS1 overexpressed group (P=0.003, P=0.006), while LY294002 could inhibit the effect produced by HMGCS1 (P=0.000). The intervention of LY294002 could reduce the expression levels of HMGCS1 and p-AKT protein and HMGCS1 mRNA (both P=0.000) in the negative control group and the blank control group. Conclusion · HMGCS1 can reduce the sensitivity of HL-60 cells to chemotherapy drug adriamycin, while PI3K/AKT signaling pathway inhibitor LY294002 can restore i

关 键 词： 羟基 甲基 辅酶 合成酶 胆固醇代谢 磷酸酰肌醇 激酶 蛋白激酶 信号通路 急性髓细胞性白血病

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