上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (9): 991-.doi: 10.3969/j.issn.1674-8115.2019.09.009

• 论著·基础研究 • 上一篇    下一篇

3-羟基-3-甲基-辅酶A合成酶1调控 PI3K/AKT信号通路影响 HL -60细胞的药物敏感性

贾妍 1, 2，王惠雯 1，易晋牟 1，曾辉 2，卢敏 3   

  1. 1. 中南大学湘雅医院血液科，长沙 410008；2. 暨南大学附属第一医院血液科，广州 510630；3. 上海交通大学医学院附属瑞金医院上海血液学研究所，上海 200025
  • 出版日期:2019-09-28 发布日期:2019-11-02
  • 通讯作者: 曾辉，电子信箱：androps2011@hotmail.com。卢敏，电子信箱：min.lu@shsmu.edu.cn。为共同通信作者。
  • 作者简介:贾妍（1992—），女，住院医师，博士生；电子信箱： jiayan1992@outlook.com。
  • 基金资助:
    国家自然科学基金（81770184）；上海市教育委员会高峰高原学科建设计划（20161305）

Effect of the PI3K/AKT signaling pathway regulatedHMGCS1 on drug sensitivity of HL-60 cells

JIA Yan1,2, WANG Hui-wen1, YI Jin-mou1, ZENG Hui2, LU Min3   

  1. 1. Department of Hematology, Xiangya Hospital, Central South University, Changsha 410008, China; 2. Department of Hematology, The First Affiliated Hospital of Jinan University, Guangzhou 510630, China; 3. Shanghai Institute of Hematology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2019-09-28 Published:2019-11-02
  • Supported by:
    National Natural Science Foundation of China, 81770184; Shanghai Municipal Education Commission— Gaofeng Clinical Medicine Support, 20161305）。

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摘要： 目的 ·探讨 3-羟基 -3-甲基 -辅酶 A合成酶 1（3-hydroxymethyl-3-methylglutaryl-CoA synthase 1，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 HMGCS1 mRNA含量，并验证 HMGCS1过表达组细胞株是否构建成功。应用 Western blotting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（ P0.000）。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（ P0.003，P0.006），而 LY294002则可抑制由 HMGCS1产生的作用（ P0.000）。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 P0.000）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。

关键词: 3-羟基 -3-甲基 -辅酶 A合成酶 1, 胆固醇代谢, 磷酸酰肌醇 3激酶 /蛋白激酶 B信号通路, 急性髓细胞性白血病

Abstract:

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 constructedinfecting 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 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 of AKT and phosphorylated AKT (p-AKT) in phosphatidylinositol 3 kinase (PI3K) / protein kinase B (PKB / AKT) signaling pathway was detectedWestern 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 was detectedqPCR and Western blotting. Results · Compared with the negative control group, the HMGCS1 mRNA was increased significantly in the HMGCS1 overexpressed group (P0.000). Compared with the blank control group and the negative control group, the p-AKT protein level in the HMGCS1 over group was significantly increased, while the AKT 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 (P0.003, P0.006), while LY294002 could inhibit the effect producedHMGCS1 (P0.000). The intervention of LY294002 could reduce the levels of HMGCS1 and p-AKT protein and HMGCS1 mRNA (both P0.000) 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 its sensitivity.

Key words: 3-hydroxymethyl-3-methylglutaryl-CoA synthase 1 (HMGCS1), cholesterol metabolism, phosphatidylinositol 3 kinase (PI3K) / protein

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