Study On The Relationship Between HIF-1α And Anti-proliferation Effect Of Evodiamine In LOVO Cells

AimTo investigate the anti-proliferation and apoptosis-inducing effects of evodiamine in human colon cancer cell LOVO, and its possible mechanisms underlying this process.MethodsCrystal violet staining, flowcytometry, luciferase reporter, RT-PCR,Western bloting has been used in the investigation. The details as following:1. The proliferation inhibitory effects of evodiamine on human colon cancer cell LOVO were analyzed with Crystal violet staining,FCM, and WB assay.2. The level of HIF-1α mRNA and protein affected by evodiamine in LOVO cells were detected with reporter, RT-PCR and WB.3. Recombinant adenovirus were used to investigate the proliferation inhibitory effect of evodiamine in LOVO cells.4. To analyze the protein level of Akt1/2, p-Akt1/2 and IGF-1.Evaluate the effect of IGF-1 on the changes of protein level of Akt1/2,p-Akt1/2 affected by evodiamine in LOVO cells.Results1. Evodiamine inhibits proliferation, arrests the cycle at S phase and promotes apoptosis.2. The transcriptional activities, mRNA and protein level of HIF-1α were decreased by evodiamine in LOVO cells.3. Adenovirus mediated HIF-1α knockdown strengthen the inhibitory of evodiamine in LOVO cells, while HIF-1α over-expression shows the opposite effects.4. Evodiamine inhibits protein level of p-Akt1/2 and IGF-1,over-expression of IGF-1 reverse the decreased protein level of p-Akt1/2 induced by evodiamine in LOVO cells.ConclusionEvodiamine showed significantly inhibitory effects in human colon cancer LOVO cells. These effects may be related with down regulating HIF-1α, which may be mediated by inhibiting PI3K/Akt signaling pathway.