Infiuence Of Artemether Combine With Thermotherapy On Tongue Squamous Cell Carcinoma Cells8113

Objective:This study was designed to investigate Whether Artemether combine with thermotherapy has Synergistic inhibitory effect to the tongue squamous cell carcinoma cells 8113 in vitro,and also to explore the possible inhibitive mechanism.Methods:(1) Grouping:control group(Tca8113),drug group(Tca8113 cells were treated with 150μM,300μM,450μM,600μM,750μM,900μM,1050μM Artemether),heating group(Tca8113 cells were treated with43℃40min,43℃80min,43℃120min),combination group(Artemether+heating).(2) Observe the changes of cell morphology by inverted microscope.(3) MTT(Methy thiazolyl tetrazolium)assay was used to evaluate the inhibitory effect of the cells.(4) Harvested cells to analysis apoptosis rate by FCM with TUNEL.(5) Harvested cells to analysis the changes of Caspase-3 activity.(6) Harvested cells to analysis the changes of mitochondrial transmembrance potential by FCM.(7) Immunocytochemistry was used to examine the expression of bcl-2 and bax in human tongue squamous Tca8113 cells (8) SPSS11.5 staistical software used for statistical analysis.Result:(1) Observation of transmission electron microscope(TEM):treated with artemether or/and thermotherapy 24h,appeared apoptotic cells:the cells became round and shrinkage. Combination group have more apoptotic cells than drug group and heating group. (2) treated with artemether or/and thermotherapy 36h,the inhibition rates of Tca8113 cells were:drug group(Tca8113 cells were treated with 150μM Artemether) is (8.64±4.61)%,drug group(Tca8113 cells were treated with 300μM Artemether) is (19.22±5.42)%,drug group(Tca8113 cells were treated with 450μM Artemether) is (33.16±5.03)%,drug group(Tca8113 cells were treated with 600uM Artemether) is (43.28±3.97)%,drug group(Tca8113 cells were treated with 750μM Artemether) is (60.17±6.92)%,drug group(Tca8113 cells were treated with 900μM Artemether) is (76.78±3.65)%,drug group(Tca8113 cells were treated with 1050μM Artemether) is (87.30±1.63)%,heating group(Tca8113 cells were treated with43℃80min) is (22.68±3.93)%,combination group (150μM Artemether+43℃80min)is (33.54±4.40)%,combination group (300μM Artemether +43℃80min) is (72.95±3.99)%,combination group(450μM Artemether+43℃80min) is (90.86±2.41)%,combination group (600μM Artemether+43℃80min) is (91.55±2.64),combination group (750μM Artemether+43℃80min) is (92.03±2.13)%,combination group (900μM Artemether+43℃80min) is (92.87±2.02) %,combination group (1050μM Artemether+43℃80min) is (93.65±1.76)%. (3) treated with artemether or/and thermotherapy 36h,apoptotic cell percentage of Groups were:control group is (2.97±1.20)%,drug group(Tca8113 cells were treated with 300μM Artemether) is (9.4±1.80)%,heating group(Tca8113 cells were treated with43℃40min) is(15.10±2.79)%,heating group(Tca8113 cells were treated with43℃80min)is (32.23±4.37)%,heating group(Tca8113 cells were treated with43℃120min)is (49.57±3.20)%,combination group (300μM Artemether+43℃40min) is (31.17±1.97)%,combination group (300μM Artemether+43℃80min) is (56.27±4.37)%,combination group (300μM Artemether+43℃120min) is (84.23±3.23) %.(4) treated with artemether or/and thermotherapy 24h,activity of caspase-3 were: control group is (0.04±0.01),drug group(Tca8113 cells were treated with 300μM Artemether) is (0.12±0.01),heating group(Tca8113 cells were treated with43℃80min) is (0.27±0.01),combination group(300μM Artemether+43℃80min)is(0.49±0.03).(5) treated with artemether or/and thermotherapy 16h, monomer percentage of JC-1 were:control group is (14.57±1.60)%,drug group(Tca8113 cells were treated with 300μM Artemether) is (20.37±1.25)%,heating group(Tca8113 cells were treated with43℃80min) is (28.40±0.50)%,combination group (300μM Artemether+43℃80min) is (60.93±0.51)%.(6) treated with artemether or/and thermotherapy 6h, the expressions of bcl-2 protein in Tca8113 were down,but Bax were up. combination group (300μM Artemether+43℃80min) is most significant.Conclusion:Thermotherapy and/or artemether could inhibit the proliferation of tongue squamous cell carcinoma Tca8113, Induced apoptosis, has Synergistic and Sensitive effect.