ABSTRACT
Objective To analysis the effect of metformin on the expression of heat shock protein and VEGF in 5637 cell line of bladder cancer. Methods 5637 cell line of bladder cancer were selected, divided into blank control group, metformin 2 mM group, 5 mM group, 10 mM group and 20 mM group, 5637 cell line of bladder cancer were treated in each group.The cell proliferation inhibition rate was detected by MTT colorimetric assay, cell clone formation rate was detected by plate clone formation test, flow cytometry was used to detect cell cycle change, the expression levels of HSP-70, HSP-90α, VEGF were detected by Western blot method.Results Compared with control group, the inhibition rate of 5637 cell line of bladder cancer after 24, 48, 72 h were higher in all concentrations of metformin group(P<0.05), the higher the concentration and the longer the action time, the stronger inhibitory effect on cell proliferation.Compared with control group, the clone formation rate of 5637 cell line of bladder cancer in all concentrations of metformin group was lower, the difference was statistically significant(P<0.05).Compared with control group, the proportion of G1 phase cells were higher, the proportion of S phase cells were lower in in all concentrations of metformin group (P<0.05), the proportion of G2 phase cells in metformin 2 mM group, 5 mM group were lower(P<0.05).Compared with control group, the expression levels of HSP-70, HSP-90α, VEGF protein were lower in all concentrations of metformin group, the difference was statistically significant(P <0.05).Conclusion Metformin has obvious inhibitory effects on the expression of HSP-70, HSP-90α, VEGF in 5637 cell line of bladder cancer, can inhibit cell proliferation and cloning, promote the occurrence of G1 block, cause cell apoptosis, the effect was dose dependent.
ABSTRACT
Objective To evaluate the effect of rapamycin on proliferation and cell cycle of human bladder cancer cell line 5637. Methods Total?ly 5637 cells were maintained in RPMI 1640 containing 10%fetal bovine serum,100 U/mL penicillin and 100μg/mL streptomycin for adherent cul?ture. Cells were grown at 37℃in a 5%CO2 incubator. The 5637 cells were treated with various concentrations of rapamycin solution(50 nmol/L, 100 nmol/L,200 nmol/L,and 400 nmol/L). The control group was daily treated with single RPMI 1640 without medication. The growth repression rate for 5637 cells was evaluated by MTT. Flow cytometry was used to investigate the effect of rapamycin of different concentrations on cell cycle of 5637 cells. Results Compared to the control group,rapamycin can inhibit the proliferation of 5637 cells in a concentration and time dependent manner. Rapamycin inhibited 5637 cells at G0/G1 thus inhibiting cell proliferation but not inducing apoptosis. Conclusion Rapamycin inhibited growth of 5637 bladder carcinoma cells and arrested cell cycle at G0/G1,indicating that mammalian target of rapamycin may play an important role in proliferation of 5637 cells and act as a new tumor therapeutic target in patients with bladder cancer.