Effects Of Akt Inhibitor Deguelin On The Growth Of Bladder Cancer Cells And Its Mechanism

Objective:To study the effects of Akt inhibitor deguelin on the growth of human bladder cancer cell lines T-24, and to elucidate the possible mechanism of such effects. And To explore whether inhibition of PI3K/Akt signal pathway increases the tumor-killing effects of anticancer drugs cisplatin and Paclitaxel on bladder cancer T-24cells. This research objective provide a new theory for Akt inhibitor deguelin targeted therapy of bladder cancer and inhibition of PI3K/Akt signal pathway enhance the chemosensitivity of bladder cancer.Methods:MTT method was used to determine growth-inhibitory effect of deguelin on human bladder cancer T-24cells, Apoptosis induced by deguelin in T-24cells was determined using DNA fragmentation assay. Wound healing test was used to measure the migration ability of T-24cells.The cell cycle was detected by flow cytometry (FCM). The expression of MDM2and GSK3β in mRNA and protein levels were detected by RT-PCR and Western blotting respectively. The inhibitory rate of T-24cells after treatment with different concentrations of individual deguelin, cisplatin, Paclitaxel and deguelin plus cisplatin, Paclitaxel were observed by MTT assay, and their cell cycle were detected by flow cytometry.Results:The deguelin singnificantly inhibited cell proliferation, blocked the cell cycle in the G0/G1phase and induced apoptosis in T-24cells. The migration capacity of deguelin-treated cells was decreased compared with that of the negative control cells. The expression levels of MDM2mRNA and protein were down-regulated, while the expression levels of GSK3β mRNA and proteins were up-regulated when T-24cell were treated by deguelin. The inhibition ratio of proliferation of T-24cells was increased significantly by cisplatin, Paclitaxel in combination with deguelin. deguelin had synergistic effects with cisplatin, Paclitaxel, Flow cytometry showed that deguelin and cisplatin, Paclitaxel blocked the cell cycle mainly in the G0/G1stage, while the combined medication lengthen the G0/G1phase and shorten the S phase compared with individual deguelin, cisplatin, Paclitaxel.Conclusion:deguelin can inhibit the proliferation and induce the apoptosis in T-24human bladder cancer cells. Moreover, deguelin can make the growth of the T-24cells blocked in G0/G1phase and decreases the migration ability of T-24cells signifycantly. These effects may be related with its function in down-regulating MDM2and up- regulating GSK3β, both of them are down-stream signal meleculars in the Akt pathway.In the other, The tumor-killing effect of Cisplatin, Paclitaxel on T-24cells was increased significantly by inhibition of PI3K/Akt signal transduction pathway.