Effect Of Tumor Stem Cell And Its Mechanism In Chemotherapy Resistance Of Laryngeal Carcinoma Hep-2 Cells

PartⅠEffect of magnetic activated cell sorting and chemotherapy agent DDP in enriching tumor stem cells of laryngeal carcinoma Hep-2 cellsObjective:To compare effect of chemotherapy agent DDP to MACS in sorting TSC of laryngeal carcinoma Hep-2 cells.Method:CD133 magnetic beads were applied to sort Hep-2 cells. Different concentrations of DDP were used to treat Hep-2 cells for 48 hours. Enrichment rate of CD133~+ cells by MACS and after DDP treatment was detected by FCM.Morphologic change was observed under inverse-phase microscope.Result:FCM showed that the sorting rate of CD133~+ cells through MACS was 64.33%,while after DDP treatment for 48 hours,CD133~+ cells was enriched significantly in each group of different DDP concentrations,with the maximal rate of 50.7%,at the concentration of 4μg/ml.There is a significant difference between MACS and each group of DDP(P＜0.01).Cells treated with DDP were abnormal in morphology.Conclusion:MACS and DDP sorting has respective advantages in enriching TSC in Hep-2 cells. PartⅡTumor stem cell and chemotherapy resistance in laryngeal carcinoma Hep-2 cellsObjective:To observe the effect of laryngeal carcinoma Hep-2-TSC in chemical drug DDP resistance.Method:The cell survival rate was detected through MTT and the expression rate of CD133~+ cells was detected through FCM after each group had been treated with different concentrations of DDP for 24 hours,48 hours and 72 hours.Result:Cell survival rate did not have the relation with concentrations of DDP in the range of experiment.Percentage of CD133~+ cells at concentration of 1μg/ml,at 24 hours was less than that in control group.Percentage of CD133~+ cells was higher than that in other groups at concentration of 4μg/ml at each check point.At the point of 48 hours,percentage of CD133~+cells was the highest.Conclusion:Rate of CD133~+ cells in laryngeal carcinoma Hep-2 cells changed after different DDP treatment of different concentrations for different time intervals.CD133~+ cells could resist the effect of chemical drug DDP.At the concentration of 2μg/ml,DDP could induce differentiation of Hep-2-TSC.PartⅢOct-4 maintains stemness of Hep-2-TSC and regulates its differentiationObjective:To detect expression of Oct-4 in laryngeal carcinoma Hep-2 cells and after different concentrations of DDP treatment for 48 hours.Method:Human laryngeal carcinoma Hep-2 cells were cultured and different concentrations of DDP were added.Expression of Oct-4 of the cells was detected through ICC after 48 hours.Result:ICC showed Oct-4 was expressed in Hep-2 cells.Expression of Oct-4 after different concentrations of DDP treatment has positive correlation with that of CD133.Conclusion:Oct-4 may be a marker of laryngeal carcinoma Hep-2 cells as CD133.Oct-4 maintains stemness of Hep-2-TSC and regulates its differentiation.