| Background:Acute leukemia is a kind of malignant clonal disease of hematopoietic stem cells.It is difficult to treat the disease with large number of patients,large age span and easy recurrence.The therapeutic effect of conventional chemotherapy and bone marrow transplantation varies from person to person.Therefore,it is urgent to seek better and more effective targeted therapy.Lipid metabolism is one of the main links in tumor metabolic remodeling,which plays an important role in regulating the occurrence and development of tumor.As a key enzyme in the metabolism of long-chain fatty acids,ACSL5(Long-chain acyl-Co A synthetase 5)participates in the metabolic regulation of fatty acids in vivo.At present,there are few studies on the dyslipidemia of the disease and the effect of ACSL5 on the proliferation and apoptosis of leukemia cells.Objective:By retrospectively analyzing the blood lipid levels of patients with acute leukemia diagnosed for the first time,the leukemia lipid metabolism related gene ACSL5 was excavated by bioinformatics analysis,so as to further explore the mechanism of ACSL5 affecting the proliferation and apoptosis of leukemia cells by regulating lipid metabolism.Methods:1.According to inclusion and exclusion criteria,246 acute leukemia patients with complete clinical data who were hospitalized for the first time from January2015 to December 2020 were enrolled as the experimental group.At the same time,109 patients with non-blood system diseases and healthy people served as the control group The baseline data and blood lipid levels of the two groups were statistically analyzed by using SPSS 25.0.2.Differential expression of metabolism-related genes in acute myeloid leukemia(AML)analyzed by bioinformatics;Analysis of the expression levels of lipid metabolism related gene ACSL5 in different tumors by GEPIA database;Analysis of the expression levels of ACSL5 in different types of acute leukemia by GEO database;The m RNA expression levels of ACSL5 gene in bone marrow samples from patients with acute myeloid leukemia were determined by reverse transcription polymerase chain reaction(RT-PCR);NB4 cell lines stably infected with sh ACSL5 and sh NC recombinant lentivirus were constructed as ACSL5 knockdown group and control group;Cell proliferation was assessed by manual counting and CCK-8 assays;Cell cycle was detected by PI single staining;Apoptosis was detected by annexin V/PI double staining;The expression levels of apoptosis-related genes were detected by RT-PCR;Apoptosis of cells treated with cytarabine was detected by Annexin V/PI double staining.Results:1.Serum triglyceride levels in patients with acute leukemia who were first diagnosed were higher than those in the control group(P < 0.001).Further classification showed that triglyceride levels increased more significantly in patients with acute myeloid leukemia compared with the control group,and the difference was statistically significant(P<0.001).2.Bioinformatics analysis suggested that lipid metabolism was abnormally active in AML,and further analysis in the GEPIA database showed that the lipid metabolism related gene ACSL5 was up-regulated in AML and associated with poor prognosis.Meanwhile,GEO database analysis showed that ACSL5 expression was up-regulated in AML and B-cell acute lymphoblastic leukemia.High expression of ACSL5 in patients with acute leukemia was also confirmed by RT-PCR(P<0.05).3.CCK-8 and manual counting results showed that the cell growth rate of ACSL5 knockdown group was significantly slower than that of control group(P<0.01).Flow cytometry showed that compared with the control group,the ACSL5 knockdown group had a relatively increased number of cells in the G1 phase and a relatively decreased number of cells in the G2 phase,whereas the change in the number of cells in the S phase was not obvious.4.Flow cytometry showed that the apoptosis rate of ACSL5 knockdown group was higher than that of control group and the difference was statistically significant(P<0.05).The detection of apoptosis related genes by RT-PCR showed that the expression level of Caspase-3 increased and the expression level of Bcl-2 decreased in the ACSL5 knockdown group compared with the control group.5.Different concentrations of cytarabine(0,0.2,0.4,0.6,0.8μmol/L)were used to treat the ACSL5 knockdown group and the control group,and the results showed that the apoptosis rate was concentration-dependent.Under the same concentration of cytarabine treatment,the apoptosis rate of ACSL5 knockdown group was significantly higher than that of control group,and the difference was statistically significant(P<0.001).Conclusion:1.Acute leukemia patients had abnormal blood lipid levels,and triglyceride levels were significantly increased.2.The lipid metabolism pathway of AML was abnormally active,and the lipid metabolism-related gene ACSL5 was abnormally expressed,which was highly expressed in AML patients with poor prognosis.3.Knockdown of ACSL5 could promote the apoptosis of AML cells,inhibit the proliferation of AML cells,and increase the sensitivity of AML cells to cytarabine. |
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