The Changes Of MTOR Signal Pathway In T Cells Of Aplastic Anemia

Objective:(1) To explore the role of mTOR signal pathway during the progress ofaplastic anemia (AA) by detecting the phosphorylated molecules, such as p-Akt, p-mTOR,p-TSC, p-P70S6K, p-S6,\p-4EBP1et al. and the upstream negative regulate factor Sirt1by flowcytometry, the downstream negative regulate factor GAS5was detected by real-timequantitative PCR (RT-PCR).(2) To further clarify the actived mechanisms in T cells of aplastic anemia, and toguide the search for new treatments and drugs.Methods:(1) To detect the expression of p-Akt, p-mTOR, p-TSC, p-P70S6K, p-S6,p-4EBP1and Sirt1in CD3+T cells of13aplastic anemia patients’ peripheral bloodsamples (AA group).14peripheral blood samples collected from volunteers were used asnegative control, leukemia cell line CEM was used as positive control.(2) To detect the expression of GAS5in CD3+T cells of27aplastic anemia patients’bone marrow samples.24bone marrow samples collected from volunteers were used asnegative control, leukemia cell line CEM was used as positive control group.(3) To compare the difference of Sirt1/p-Akt, Sirt1/p-TSC, Sirt1/p-mTOR,Sirt1/p-4EBP1, Sirt1/p-P70S6K, Sirt1/p-S6between the AA group and the negative controlgroup. Analyse the correlation of absolute neutrophil count (ANC) and lymphocyte count(Ly) with the ratios.Results:(1) The espression of p-Akt, p-TSC, p-mTOR, p-P70S6K, p-S6, p-4EBP1in CD3+T cells of aplastic anemia patients were (8.77±1.61),(45.05±25.53),(15.97±11.20),(17.81±11.22),(19.79±9.05),(140.60±86.05), respectively. The espression of p-Akt, p-TSC,p-mTOR, p-P70S6K, p-S6, p-4EBP1in CD3+T cells of the control group were (2.83±1.04),(16.65±10.84),(2.95±1.23),(3.55±1.46),(2.96±1.01),(26.83±16.82), respectively. So, the expression of the phosphorylated molecules in aplastic anemia patients were obviouslyhigher than the control, P<0.05.(2) The espression of Sirt1in CD3+T cells of aplastic anemia patients was (9.08±2.10)which had no significant with the control group (10.39±2.31), P=0.149. But the ratios ofSirt1/p-Akt, Sirt1/p-TSC, Sirt1/p-mTOR, Sirt1/p-4EBP1, Sirt1/p-P70S6K were muchlower than the negative control group, P<0.05. There were positive correlation betweenANC and the ratios of Sirt1/p-TSC, Sirt1/p-mTOR, Sirt1/p-4EBP1, Sirt1/p-P70S6K andnegative correlation between Ly and the ratios of Sirt1/p-mTOR, Sirt1/p-4EBP1(P<0.05).The expression of GAS5in the aplastic anemia patients was (9.04±4.26) which was muchlower than the control group (12.43±4.50), P<0.05.(3) In the positive control group cell line CEM, The espression of p-Akt, p-TSC,p-mTOR, p-P70S6K, p-S6, p-4EBP1were (19.50±4.72),(103.9±20.09),(31.64±17.37),(35.36±15.35),(46.78±14.70),(264.74±42.01), which were all obviously higher than theAA group.(4) In the positive control group cell line CEM, the expression of Sirt1was(9.24±2.53), there were no obvious difference between the positive control group and theAA group, P>0.05. The expression of GAS5in the positive control group was (2.89±1.41)which was lower than the AA group, P<0.05.Conclusions:(1) The phosphorylated molecules in the AA group were significantlyhigher than the negative control group, but lower than the positive control group. whichsuggested that the mTOR signal pathway in T cells of aplastic anemia was in relativeactivation, and likely to be involved in the activation of T lymphocytes in AA group.(2) The expression of Sirt1in AA group relatively insufficient. And the downstreamregulator GAS5in AA group was obviously lower than the negative control group, whichsuggested the lack of GAS5and Sirt1further promote the activation of T lymphocytes inAA group.(3) The ratios of Sirt1with the phosphorylated molecules of mTOR signal pathwayshowed: the mTOR pathway in AA group migrated than the normal, which resulted inincreased lymphocyte percentage and inhibition of hematopoiesis.(4) The mTOR signal pathway was activated in the acute T lymphocyte leukemia cellline CEM, and the insufficient expression of GAS5and Sirt1may be involved in theleukemia development.