Inhibitive Effect of Chonglou Recipe on Proliferation of Human Gastric Cancer Cell Line MKN-45 and Its Possible Molecular Mechanism

Objective To investigate the effect of Chonglou Recipe(CP) on proliferation,cell cycle and aopotosis of human gastric cancer cell line MKN-45,and to explore the possible molecular mechanism.Methods After treating human gastric cancer cell line MKN-45 with CP at various concentrations(0.025～1.6 mg·mL-1) for 48 h,the proliferation rate of cell line MKN-45 was detected with methyl thiazolyl tetrazolium(MTT) method.The apoptosis and cell cycle phases were examined by flow cytometery(FCM).Reversed transcriptase polymerase chain reaction(RT-PCR) method was used to detect the mRNA expression of survivin,c-IAP1,c-IAP2 and Bcl-2.Results CP inhibited the proliferation of human gastric cancer cell line MKN-45 in a dose-dependent manner.The median effective concentration IC50 after incubation with CP for 48 hours was 0.67 mg·mL-1.After treatment with CP at the dosage of 0.2,0.4,and 0.8 mg·mL-1 for 48 hours,the apoptotic rate of cell line MKN-45 was 7.0 %,13.8 % and 36.3 %,respectively,showing a dose-dependent trend.The apoptotic rate was 3.4 % in the control group.After treatment with CP at the dosage of 0.8 mg·mL-1 for 24 h,the percentage of MKN-45 at G0/G1 phase was 73.16 %,and was 63.30 % in the control group,which was higher than the percentage of MKN-45 at S phase(22.52 % in CP group and 32.48 % in the control group).After MKN-45 cells were exposed to CP at the indicated concentration of 0.8 mg·mL-1 for 48 h,the mRNA expression of survivin,c-IAP1,c-IAP2 and Bcl-2 were down-regulated.Conclusion CP can inhibit the proliferation and induce apoptosis in human gastric carcinoma cell line MKN-45.Disturbing the cell cycle and down-regulating the mRNA expression of survivin,c-IAP1,c-IAP2 and Bcl-2 may be one of its therapeutic mechanisms.