The Molecular Mechanisms Of Long Non-coding RNA MANTIS Affect The Growth Of Acute Myeloid Leukemia

Objective:Acute myeloid leukemia(AML)is a common malignant tumor in the blood system.It can be treated with high dose chemotherapeutic drugs to consolidate chemotherapy or stem cell transplantation to prolong the survival of patients,but the prognosis of acute myeloid leukemia is still very poor in recent years.It has been found that the pathogenesis of AML is related to many factors,such as recurrent somatic mutation and gene expression,but the molecular mechanism of AML is not clear,so the molecular mechanism of AML is deeply studied.In order to obtain specific biomarkers or new therapeutic targets,we can accurately predict the prognosis of patients with AML and choose the most appropriate treatment.It has been shown that long noncoding RNA plays an important role in the process of proliferation,invasion and metastasis of many kinds of tumors by taking part in the regulation of many signal pathways and biological functions,which can induce angiogenesis and play an important role in the process of proliferation,invasion and metastasis of many kinds of tumors.Play a role similar to oncogenes or tumor suppressor genes.In the hematopoietic system,lnc RNA is involved in the maturation and differentiation of blood cells,and is associated with the occurrence of many malignant hematological diseases,such as leukemia.lnc RNA MANTIS is a new type of differentially regulated lnc RNA with the function of promoting endothelial angiogenesis.The purpose of this study was to investigate the expression of MANTIS in AML and the role of MANTIS in the proliferation and apoptosis of U937 cells.To explore whether it is involved in Notch signaling pathway.Methods: 1 36 patients with acute myeloid leukemia and 11 volunteers were selected to collect and isolate bone marrow mononuclear cells to detect the expression of lnc RNA MANTIS by Real-time PCR;2 Cell culture;3 Overexpression pc DNA-MANTIS was transfected into U937 cells using Lipofectamine 3000(Invitrogen);4 The expression of MANTIS in overexpression pc DNA-MANTIS U937 cells was detected by Real-time PCR;5 Cell proliferation was measured by CCK-8 Cell Counting Kit;6 Apoptosis detection by Annexin ?-FITC/PI double stainning and flow cytometry analysis;7 Protein analysis by Western blot;8 Statistical analysis: The data are expressed as means ± Standard Deviation and the difference between two groups was evaluated using Student's test.A probability level of 0.05 is used to establish significance and indicated as P<0.05.Results: 1 Real-time PCR method was used to detect the relative expression of MANTIS in AML patients.The results showed that the expression of MANTIS in AML patients was significantly lower than that in normal controls,and there was statistical significance between the two groups(P<0.01).2 Real-time PCR method was used to detect the relative expression of MANTIS in in U937 cell line.The results showed that the expression of MANTIS in AML was significantly lower than that in normal controls,and the two groups had statistical significance(P<0.05).3 The expression of MANTIS in overexpression pc DNA-MANTIS U937 cells was detected by Real-time PCR that the MANTIS expression increased significantly compared with the control group,and the two groups had statistical significance(P<0.05).4 The cell viability of U937 cells transfected with pc DNA-MANTIS were greatly decreased compared with that of U937 cells transfected with pc DNA.The cell proliferation inhibition rate of U937 cell transfected with pc DNA-MANTIS for 12 h,24h,48 h and72h is(11.16±2.13)% ?(15.47±0.31)% ?(21.86±1.71)% and(42.97±2.61)% respectively;There is a statistical difference compared with the control group(P<0.05).5 U937 cells transfected with pc DNA-MANTIS could induce cell apoptosis.The apoptotic rates of U937,U937/vector and u U937/MANTIS were(31.71 + 0.32)%,(35.42 + 1.94)% and(59.06 + 2.37)%,respectively,The difference was statistically significant(P<0.05).6 To investigate the regulated effect of MANTIS on the Notch1 signaling pathway,U937 cells were transfected with pc DNA-MANTIS or vector pc DNA,then the protein level of Notch1 and DLL4,Hes1 were detected.Overexpression of MANTIS increased the protein level of Notch1 and DLL4,Hes1.There is statistically difference compared between the two groups(P<0.05).Conclusions: 1 Low expression of MANTIS was observed in the acute myeloid leukemia bone marrow blood and cells.Exogenous upregulation of MANTIS expression in U937 cell line can inhibit the proliferation and induce apoptosis.2 Exogenous upregulation of MANTIS expression in U937 cell line canincreasedthe protein level of Notch1 and DLL4,Hes1.