Abstract
Objective To study the apoptotic activity of hydroxycamptothecin (HCFT) against human gastric carcinoma and its mechanism of action.Methods The apoptotic activity of HCPT against gastric carcinoma SGC-7901 cells was detected by MTT assay.The apoptosis occurrence was determined by Annexin V/PI staining and flow cytometry analysis.The mRNA expression of Bmi-1 was detected by real-time PCR analysis.Western blot analysis was used to detect the protein expression of Bmi-1.Results The IC50 values of HCPT against gastric carcinoma SGC-7901 cells was (4.87 ± 0.35) μ mol/L.After treatment with 0,5,10,20 μ mol/L HCPT for 48 h,the percent of apoptosis was elevated from (1.62 ± 0.37)% to (21.45 ±4.54)%,(36.67 ±5.38)%,(54.26 ±7.14)% and the 2-△Ct values of Bmi-1 mRNA was decreased from 0.614 ±0.022 to 0.445 ±0.018,0.376 ±0.012,0.215 ±0.010,there had significant difference among the different treatment (P < 0.01).Treated with 0,5,10,20 μ mol/L HCPT for 48 h,the protein expression of Bmi-1 was down regulated by HCPT treatment in a dose-dependent manner by Western blot analysis.Conclusion HCPT shows effective activity to induce apoptosis in gastric carcinoma cells,which is mainly related to the down regulation of Bmi-1. Key words: Stomach neoplasms; Apoptosis; Camptothecin; Polycomb gene Bmi-1
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