Study On Reversal Of Multidrug Resistance In Oral Squamous Cell Carcinoma Tca8113 Cells By Artemisinin

Objective: Through establishing a human oral squamous cell carcinoma Tca8113 cells model of carboplatin(Ka),a chemotherapeutic drug by addition in vitro,to observe the expression of drug resistance-related genes P-gp and GST-π in human oral squamous cell carcinoma Tca8113 cells resistant of carboplatin(Ka),and then using Chinese medicine artemisinin to intervene oral squamous carcinoma Tca8113 cells resistant of carboplatin(Ka).The changes of P-gp and GST-π were compared before and after and the changes in the expression of corresponding proteins were compared before and after too.Finally,to investigate reversal of multidrug resistance in oral squamous cell carcinoma Tca8113 cells by artemisinin.Method: 1.Establishment of drug-resistant cell line of human oral squamous carcinoma Tca8113 by induction in vitro.The chemotherapy drug used in this experiment was carboplatin(Ka),and the induction method was increasing the drug gradually by small doses of long duration exposure.2.The expression of resistance related genes P-gp and GST-πin drug-resistant cell lines was detected by immunocytochemical method.3.Tca8113 cells resistant to carboplatin(Ka)were cultured in vitro.With different concentrations of artemisinin(0 μg/ml,12.5 μg/ml,25 μg/ml,50 μg/ml,100 μg/ml)on resistant cells(24hours,48 hours,72hours),after that,the effect of artemisinin on the proliferation of drug-resistant cells was detected by CCK8 method.And the changes of P-gp and GST-π in different concentrations of artemisinin(0 μg/ml,12.5 μg/ml,25 μg/ml,50 μg/ml,100 μg/ml)in drug-resistant cells were detected by RT-PCR.The changes of protein expression levels of P-gp and GST-π in different concentrations of artemisinin in the drug-resistant cells were detected by Western-blot.Results: 1.The immunocytochemistry results showed that the positive expression rate of P-gp and GST-π in Tca8113 cells resistant of carboplatin(Ka)was much higher than that of Tca8113 cells,and it was thought that the expression of P-gp and GST-π was increased in Tca8113 cells resistant of carboplatin(Ka)than in Tca8113 cells(P=0.018,P=0.020,P<0.05).2.The CCK8 cell proliferation experiment results showed that artemisinin could inhibit the proliferation of Tca8113 cells resistant of carboplatin(Ka),and different concentrations of artemisinin intervention group compared with the control group,the difference was statistically significant(P=0.000,P<0.001);With the increase of concentration of artemisinin,the inhibition rate of cell proliferation was also increased,and the inhibition rate reached the maximum when the concentration of artemisinin was 100μg/ml.Under the same artemisinin was 100 μg/ml,the inhibition rate was also increased with the increase of time,and the inhibition rate reached the maximum at 72 hours.(P=0.000,P<0.001).3.RT-PCR results showed that compared with the resistant group,the mRNA expression of both P-gp and GST-π mRNAs decreased gradually with the increasing concentration of artemisinin(P<0.05).Different concentrations of artemisinin reversal group compared with non drug resistant group,the differences were statistically significant(P-gp gene F=5.670,P=0.000;GST-π gene F=2.108,P=0.000).4.Western-blot results showed that compared with the resistant group,the protein expression levels of P-gp and GST-π decreased in the reversed group treated with artemisinin(P<0.5).Different concentrations of artemisinin reversal group compared with non drug resistant group,the differences were statistically significant(P-gp gene F=206.661,P=0.000;GST-π gene F=153.016,P=0.000).Conclusion: 1.Artemisinin can inhibit the proliferation of drug-resistant oral squamous carcinoma Tca8113 cells,and its inhibitory effect increases with the increasing concentration of artemisinin.2.Under the action of artemisinin,resistant to carboplatin(Ka)reduced the expression of P-gp and GST-π in oral squamous carcinoma Tca8113 cells,thus reversing the drug resistance of drug-resistant cells.3.Artemisinin in the treatment of oral squamous cell carcinoma may have potential clinical research value.It may open up new fields for the types of oral squamous cell carcinoma,new anticancer drugs and adjuvant chemotherapy drugs.