Effect of the PI3K/AKT signaling pathway regulated by HMGCS1 on drug sensitivity of HL-60 cells / 上海交通大学学报（医学版）

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 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 its sensitivity.