The Studies To Anti-tumor Effect And Its Mechanisms Of Nsaids Combined With Sodium Butyrate On Human Colon Cancer Cell Line HCT-116 In Vitro

Objective: To study the anti-tumor effect and its mechanisms of NSAIDs combined with sodium butyrate on human colon cancer cell line HCT-116 in vitro.Methods: We subculture the colon cancer cell strain HCT - 116, adjust cell to logarithmic growth period. Take logarithmic growth period cells to 96-well plates , make cell concentrations reach 1.0 x 104 each hole , 100ul each hole, cultivate three days later, adds celecoxib 100umol/L, 200umol/L, 400umol/L, sodium butyrate 0.5mmol/L, 1mmol/L, 2mmol/L, celecoxib 100 umol/L plug 0.5mmol/L sodium butyrate. After treatment by the different factors,we continue to cultivate cells 24h,48h,72h.Then we use MTT method analyze the effect of each drug on cells; application flow cytometry detect cell cycle distribution; application RT-PCR detect the express of COX - 2 and hTERT in colon cancer cell HCT-116 and the influence of non-steroidal anti-inflammatory drug and sodium butyrate to express of COX-2 and hTERT.Results:1. The MTT method shows celecoxib, sodium butyrate inhibit colon cancer cell HCT–116 growth significantly, with the increase of concentration and the extension of time, the effect of inhibition enhanced obviously (P < 0.05), the inhibition effect of combination celecoxib with sodium butyrate enhanced obviously than that adds them alone (P < 0.05).2. After colon cancer cell HCT–116 was intervention by celecoxib or sodium butyrate alone 48h,,the ratio of G0/G1 phase was increased and the ratio of S phase was decreased . With drug concentration increases, the effect get more obviously . Comparing with celecoxib or sodium butyrate alone,G0/G1 phase was increased and S phase was decreased significantly by celecoxib combined with sodium butyrate. By statistical analysis, compared with the control group,G0 / G1 phase increase, S phase decrease in all medicine-consumption group,the difference was statistically significant (P < 0.05), combined treatment group have statistically significant than single-agent group (P < 0.05).3. COX - 2mRNA and hTERTmRNA of colon cancer cell HCT-116 are high expression, RT - PCR products show positive strips by agarose gel electrophoresis , compared with standard DNA Marker, we can realize the strip at 329bp represent hTERTmRNA and the strip at 305bp represent COX-2mRNA。Compared with the control group,with the celecoxib concentration increased, COX - 2mRNA express decrease gradually,the difference was statistically significant (P < 0.05). Sodium butyrate group also presenting the same characteristics. Comparing with celecoxib or sodium butyrate alone, COX - 2mRNA express was decreased significantly by celecoxib combined with sodium butyrate (P < 0.05). Compared with the control group,with the celecoxib concentration increased, hTERTmRNA express decrease gradually,the difference was statistically significant (P < 0.05). Sodium butyrate group also presenting the same characteristics. Comparing with celecoxib or sodium butyrate alone, hTERTmRNA express was decreased significantly by celecoxib combined with sodium butyrate (P < 0.05).Conclusion:1 The growth of HCT-116 cell in vitro was inhibited by celecoxib or sodium butyrate and in dose-and-time dependent manner.The anti-carcinoma effect was significantly enhanced by celecoxib combined with sodium butyrate,and maybe its mechanisms were arresting cell cycle。2. THE mechanisms of celecoxib or sodium butyrate inhibit cell growth should be inhibit the expression of COX-2 and telomerase in transcription level, which block the cell cycle and apoptosis.