Effect of 3-MA on arsenic trioxide induced apoptosis in Jurkat cells

Objective: To discuss the effect and mechanism of autophagy inhibitor 3-MA on arsenic trioxide inducing apoptosis of acute T-cell leukemia cell line Jurkat cells. Methods: Proliferation inhibition of Jurkat cells treated with arsenic trioxide was detected by XTT. Morphological characteristics of Jurkat cells treated with different concentrations arsenic trioxide were observed by electron microscope. Microtubule-associated protein 1 light chain 3B( LC-3B) protein expression was detected by Western blot and flow cytometry. Apoptosis rates of Jurkat cells treated with 3-MA combining arsenic trioxide were detected by flow cytometry using AnnexinV-FITC /PI double staining. Results: Arsenic trioxide inhibited the growth of Jurkat cells in a dose and time dependence. We observed different morphological characteristics of autophagy,apoptosis and necrosis accompanying more autophagosomes in Jurkat cells which were treated with arsenic trioxide 2. 5,5,10 μmol /L after 24 h. LC3 B mean fluorescence intensity( MFI) relative multiples were( 3. 1 ± 0. 2)fold,( 4. 6 ± 0. 31) fold,( 34. 2 ± 4. 5) fold with 5 μmol /L arsenic trioxide treated Jurkat cells 0,24,48 h,and the P values between each of the two groups were less than 0. 05,which increased depending time consistently with the growth inhibition rates. LC-3B protein expression gradually increased treated Jurkat cells with arsenic trioxide after 24 h,48 h. The growth inhibition rate( 60. 6 ± 8. 3) % was significantly different treated with arsenic trioxide combining 3-methyl adenine( 3-MA) while it was( 33. 4 ± 9. 1) % treated with arsenic trioxide alone,however,LC-3B protein expression gradually decreased. Jurkat cell apoptosis rate( 44. 96 ± 3. 60) % was significantly increased treated with arsenic trioxide combining autophagy inhibitor( 3-MA) while it was( 2. 94 ± 0. 26) % treated with arsenic trioxide alone,and this difference was statistically significant. Conclusion: 3-MA increased apoptosis rates of Jurkat cells inducing by Arsenic trioxide and it may be related with inhibition of autophagy and induction of apoptosis.