Effect Of Arsenic Trioxide On Inducing Apoptosis Of T Cell Lymphoma Cell Line Hut-78 Cell

Objective: To investigate the effect of Arsenic Trioxide on T cell lymphoma cell line Hut-78 cells and the possible function and mechanism by detecting the proliferation, induce apoptosis and cell cycles effects of T-cell lymphoma cell line which was treated with Arsenic Trioxide of defferent concentrations.Methods: The inhibitory effect of Arsenic Trioxide on proliferation and its inducing apoptosis effect, influence on cell cycle of hut-78 cell line was deteced by MTT, Flow cytometry with PI staining and terminal deoxynucleotidy transferase-mediated dUTP nick end-labeling(TUNEL) assay. The transcription level changes of bcl-2 gene and VEGF gene was measured by RT-PCR.Results: 1. In the concentration range of 2 - 10μm/L of As₂O₃, with the increasing of concentration and extending of action time, the growth inhibition of Hut-78 cells was gradually increased. At the level of 2μmol / L concentration for 24 hours, the inhibition rate was lowest in each time group, and at the level of 10μmol / L for 72 hours the inhibition rate of Hut-78 cells reached to 64.16±6.65%. 2. Flow cytometry analysis of DNA content showed that, the sub-diploid peak of Hut-78 cells were treated with As₂O₃ for 24 hours was not obvious, but after 48 and 72 hours the sub-diploid peak was significant appeared. 3. To investigate the cell cycle change of Hut-78 cells was treated with Arsenic Trioxide, we used the Flow cytometry with PI staining. The results showed that the ratio of G0-G1 phase of Hut-78 cells which was treated without Arsenic Trioxide was higher, the ratio of G2-M phase of Hut-78 cells which was treated with Arsenic Trioxide was increased. When the Hut-78 cells was treated with 10μmol/L Arsenic Trioxide for 72 hours, the highest percentage of G2-M phase. was reached . 4. To investigate the apoptosis of Hut-78 cells was treated with Arsenic Trioxide, we used the Tunel method. The results showed that, the brown apoptotic cells was appeared when the Hut-78 cells was intervented with 2-10μmol / L As₂O₃ for 48 or 72 hours, .and with the increasing of concentration and extending of action time, the effect was gradually changed . Rate of apoptosis and necrosis were lowest at 2μmol/L for 48 hours ,but reached a maximum at 10μmol/L.for 72 hours 5. we used the RT-PCR method to observed the expression level change of Bcl-2 gene and VEGF gene of Hut-78 cells which was treated with Arsenic Trioxide in different concentration for different action time. It showed that Bcl-2 gene and VEGF gene expression decreased gradually with the concentration of As₂O₃ increasing and acting time extending.Conclusion: In a certain range of concentrations, arsenic trioxide can inhibit proliferation of Hut-78 cells and induce apoptosis of Hut-78 cells, and it was depended by both time and dose. It may be raleted to the transcription level of bcl-2 gene and VEGF gene decreased.