| Objective:The study focuses on the relationship between the effect of I3C on apoptosis and the expression of p53/MDM2 protein in human laryngeal carcinoma Hep-2 cells. Therefore, it will provide a theoretical basis for the mechanism of I3C on laryngeal carcinoma and the drug action screeningMethods:(1) Culture of Hep-2 cell.(2) Cell vitality rate of Hep-2 cell treated with different concentration of I3C (0μM,75μM,.150μM) at various time point (Oh, 12h,24h,72h) was determined through the MTT assay.(3) The mRNA levels of p53, MDM2, BAX and Bcl-2 treated with 150μM I3C after 72h were investigated through semi-quantitative RT-PCR method.(4) The protein levels of MDM2, Bcl-2, Bax and p53 were investigated through Western Blotting method.Results:1 Compared with the control group (TO), cell vitality rate of I3C group decreased as the time elapsed when different concentration of drugs were given to Hep-2 cells (P<0.05); and cell vitality rate of the high concentration group (150 M) was lower than that of the low concentration group (75 M) at the same time point (P<0.05).2 The PCR results showed that the intracellular expression of MDM2 and Bcl-2 mRNA reduced by using 150M I3C on Hep-2 cells after 72h comparing with the control group (P<0.05) while the expression P53 and BAX mRNA increased (P<0.05).3 The Western Blotting results showed that the intracellular expression of MDM2 and Bcl-2 protein decreased by using 150M I3C on Hep-2 cells after 72h comparing with the control group (P<0.05), however, the expression of BAX and P53 protein increased (P<0.05).Conclusion:1.Inhibitory effect of I3C on the proliferation of Hep-2 cell is time-dependent and concentration dependent.2.13C upregulates BAX/Bcl-2 and then promotes the apoptosis of laryngeal carcinoma cells.3.13C downregulates ubiquitination of p53 to restrain the proliferation of laryngeal carcinoma Hep-2 cells. |
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