Combination Effect of ATO/THAL On PI3K/AKT/ Mtor Pathway in AML Cell Lines

Background: Acute myeloid leukemia (AML) is a heterogene-ous group of malignant that is caused by an uncontrolled pro-tein of clonal neoplastic cells and its accumulation in the bone marrow.AML resistant cell line (CD34+) is 10-15 times more susceptible than sensitive cell line (CD34-) to chemotherapy Anticancer effect. PI3K/AKT/mTOR signaling cascade is cru-cial to many widely different physiological processes such as cell survival and apoptosis. The previous studies demonstrated the cytotoxic activity of Arsenic trioxide (ATO) in many can-cers. Thalidomide (THAL) is also an angiogenesis inhibitor it blocks the survival in cells. Therefore, investigating the effect of ATO and THAL on PI3K/AKT/mTOR and PTEN mRNA expression is the primary goal of this study.Materials and Methods: KG-1 and U937 cells (respectively as resistant and sensitive cell lines to chemotherapy) treated with ATO and THAL with different dose and time manner. Cell proliferation was evaluated by MTT assay. The rate of apopto-sis was measured by flow cytometry (Annexin-V/PI) also cell cycle analysis was done by flow cytometry (PI). Furthermore, the effect of cited compounds on the mRNA expression level of PI3K/AKT/mTOR and PTEN were measured by Real Time-PCR.Results: Effective dose and IC50 of THAL in KG-1 and U937 respectively were 80µM and 60µM. Effective dose and IC50 of ATO in KG-1 and U937 respectively were 1.618µM and 1µM. Annexin-V/PI staining indicated that ATO and THAL can in-duce apoptosis in both cell lines. Cell cycle analysis showed that cells arrested in subG1/G1 in the presence of ATO/ THAL. Results of Real Time-PCR showed that the level of mRNA Ex-pression of PI3K/AKT/mTOR was decreased and PTEN was increased in both cell lines.Conclusion: According to the obtained results combination of THAL and ATO has an anti-leukemic effect by downregulating of PI3K/AKT/mTOR and upregulating of PTEN.

کلیدواژه ها:

AML ، Arsenic Trioxide ، Thalidomide ، PI3K