Detection Of FOXM1the Acute Leukemia Cells And The Relationship With The Antitumor Activity Of Metformin

Objective：We investigated the expressions of FOXM1mRNA in untreated acute leukemia (AL)patients, and analyzed the correlation with the patients’ prognosis. We tested theexpression of FOXM1in ML-2cells and Jurkat cells, after we have incubated themwith Metformin. The expressions and the relationship of FOXM1and mTOR weredetected after the ML-2cells were treated with Thiostrepton. The effect of Metforminon the apoptosis, cell cycle and proliferation of ML-2cells were tested.Method:1. Total mRNA was extracted from the untreated AL patients. After it was reversedto cDNA, the expression was tested through RT-PCR. We got the blood routine,bone marrow cytology, chromosome and gene detection data, aiming to discoverwhether the FOXM1expressed had the relationship with them. Then we assesswhether the FOXM1expressed means a poor prognosis.2. After having been incubated with Bortezomib for48h, the Jurkat cells and ML-2cells’ FOXM1expression was tested. Having treated the Jurkat cells with theFOXM1inhibitor Thiostrepton for16h, we tested the expression of FOXM1andmTOR to find whether their changes have relations.3. We treated ML-2cells with Metformin, then detected the expressions of FOXM1mRNA with RQ-PCR and the protein expression with Western Blot.4. After the ML-2cells having been incubated with Metformin for24h and48h,Flow CytoMeter stained Annexin V-PI was applied to detected the apoptosis, PIwas applied to tested the cell cycle. CFSE labled cell proliferation trail was servedto check out the proliferation changes.Results: 1. This study analyzed71patients with primary AL, the positive rate was85.92%. But there was no significant relation with the peripheral white blood cells, bone marrow blast cells percentage, and genetics prognosis level.2. After having been incubated with Bortezomib for48h, the Jurkat cells and ML-2cells’ FOXM1expression was depressed on dose-dependent. The expressions of FOXM1and mTOR were depressed after the Jurkat cells having been treated with Thiostrepton for16h. Their expressions have significant relation.3. The expressions of FOXM1mRNA, but it had no relationship with the dose. The protein level were depressed on dose-dependent manners.4. After the ML-2cells having been incubated with Metformin for24h and48h, the apoptotic ratio were increased in2μmol and8μmol groups. The ML-2cells’ cycle were arrested in G0/G1and G2/M, causing the S cell cycle decreased in the2μmol and8μmol groups after they having been treated for24h. The proliferation ability was remarkably inhibited after incubated with Metformin for24h in2μmol and4μmol groups.Conclusions:FOXM1gene expression was higher in AL patients than healthy persons. Bortezomib inhibit the expression of FOXM1in ML-2and Jurkat cells. The inhibition of Thiostrepton to the expression of FOXM1causes the depressed expression of mTOR. The two genes have significant correlation. FOXM1may be an attractive target for cancer therapy. Metformin down regulates the expression of FOXM1in both mRNA level and protein level. Metformin promotes the apoptosis of ML-2cells, inducts G0/G1and G2/M cell cycle arrest, and inhibits the proliferation. It may be a new option in leukemia treatment.