The Preliminary Mechanism Of β-arrestin1 Via MiR-652-5p Regulating Glycolysis In Jurkat Cell Lines

Objective: Acute lymphoblastic leukemia(ALL)is a malignant hematological malignancy that originates in the lymphatic system and is one of the common tumors in children.Presenting symptoms of ALL include pale and fatigue from anemia,infection caused by neutropenia,bleeding due to hrombocytopenia and leukemic infiltration of liver,spleen,lymph nodes,mediastinal and central nervous system infiltration.ALL is divided into T-cell acute lymphoblastic leukemia(T-ALL)and B-cell acute lymphoblastic leukemia(B-ALL).Although T-ALL only accounts for approximately 10% to 15% of children with ALL,the survival rate of B-ALL has reached 90%,the cure rate of T-ALL is only 75%,the cure rate of refractory T-ALL and relapsed T-ALL are worse,and the new mechanism of T-ALL needs to be explored.Methods: Lentivirus were used to construct Jurkat cell lines with impaired β-arrestin1 and miR-652-5p gene,lactate production was measured by colorimetric assay,the production of ATP and ADP was detected by ATP/ADP kit.To detect the glycolysis levels in different Jurkat cell lines,the expression of hexokinase 2(HK2),glucose transporter 1(Glut1)and lactate dehydrogenase A(LDHA)involved in glycolysis were detected by RT-PCR;Bone marrow of children diagnosed with T-ALL were collected from 2017 to 2018,the patient samples were analyzed by transcriptome sequencing,the expression of β-arrestine1 and miR-652-5p were detected by RT-PCR,statistical analysis of the correlation between β-arrestin1 and mi R-652-5p expression was performed by SPSS;The target genes of miR-652-5p were predicted via mi RDB database and were confirmed by RT-PCR and western blot.Results: Jurkat cells lines were successfully constructed with impaired β-arrestin1,compared with the control cell lines(Jurkat Scram),the lactate production of Jurkat Siβ1 was increased(P<0.001),up-expression of glycolytic key enzyme HK2 was found in Jurkat Siβ1(P<0.05).Down-expression of β-arrestine1(P<0.01)and up-expression of miR-652-5p(P<0.05)were observed in patients with T-ALL.Compared with the control cell lines(Jurkat Scram),the expression of miR-652-5p in Jurkat Siβ1 was increased(P<0.001),and the expression of β-arrestine1 and miR-652-5p was negatively correlated.Jurkat cells lines were successfully constructed with impaired miR-652-5p,compared with the control cell line(Jurkat Mock),the lactate production of Jurkat Down was decreased(P<0.05).Compared with the control cell line,the expression of miR-652-5p target gene OTUD7 B was increased in Jurkat Down cell line(P<0.05),and decreased in Jurkat Siβ1 cell line(P<0.05).Conclusion: β-arrestine1 paly a role in the glycolysis of T-ALL,and its possible mechanism may through regulating the expression of miR-652-5p,which complete the mechanism of β-arrestin1 regulating the glycolysis of leukemia.