Correlation Of Autophagy On The Effect Of As₂O₃Inhibiting Jurkat Cells

BackgroundAutophagy is a prevalent pathological physiological mechanism in normal cells and tumor cells. Autophagy activity is low in some cancer cells which is related with tumorigenesis. Anticancer drugs can induce autophagy in cells and involve related molecules of the regulation of autophagy, but it may also lead to apoptosis. Autophagy has both positive and negative impact on tumor cells, and it has a broad prospects as a target for anticancer drugs. Arsenic was first used in clinical treatment of acute promyelocytic leukemia (APL) as anti-leukemia drug. And it has been used to treat multiple myeloma, myelodysplastic syndrome and various solid tumors, but its exact mechanism is still not clear.ObjectiveInvestigate the the effect of autophagy inhibitors3-methyl adenine (3-MA) and arsenic trioxide (As2O3) in acute T cell leukemia cell line. The mechanism of apoptosis provides the evidence for further study about the development of autophagy in acute T-cell leukemia.Methods1. As2O3can inhibit the growth of Jurkat cells of acute T lymphoblastic leukemia cell line, and this effect is dose-and time-dependent manner.2. Cell morphology was observed under the electron microscope after24h of different concentrations of As2O33. detect microtubule-associated protein1light chain3B (LC-3B) protein expression with Western blot and flow cytometry. 4. Detect apoptosis of3-MA on arsenic trioxide in acute T-cell leukemia with AnnexinV-FITC/PI double staining and flow cytometry.Results1. Arsenic trioxide inhibited the growth of Jurkat cells in a dose and time dependence but not obvious,2. Different forms of autophagy, apoptosis and necrosis can be observed in the electron microscope.3. After5μmol/L As2O3treated Jurkat cells, LC-3B mean fluorescence intensity were3.1±0.2-fold,4.6±0.31-fold,34.2±4.5-fold, the differences between the two groups was statistically significant (P<0.05), with growth inhibition in a time-dependent manner; immunoblotting also showed LC-3B protein expression was increased gradually.4. After24hours treatment with arsenic trioxide, the apoptosis cells greatly increased but did not obviously change in Arsenic trioxide group compared with control group.ConclusionAutophagy inhibitor3-MA can increase apoptosis rate of arsenic trioxide (As2O3)-induced in Jurkat cells, the mechanism is closely related.with induction of apoptosis and inhibition of autophagy...