| Objective: Uterine cervix cancer damages women'health seriously. In late years, the position of chemotherapy has been promoted continuously. And cisplatin is the first choice in chemotherapeutics. But great dose of drugs brings serious adverse reactions and drug fast which comes out as a challenge to the therapeutic action. As a result, it is the key to decrease the doses of chemotherapeutics and overcome drug fast in order to raise 5-year-survival rate of advanced uterine cervix cancer. Therefor, it seems extremely impotant to find some way to increase chemotherapeutics'sensitivity.This experiment determines thilidomide (Thd) only and combined with cispline (DDP) the inhibition effect of Hela cells'growth in out-of-body condition. Using MTT method analyzes the influences of cell cycle and apoptosis with flow cytometry (FCM), which aims at approaching whether thalidomide can inhibit Hela cells'growth out of body or promote Hela cells'apoptsis, the combined effection with cisplatin and the influences on ant-apoptsis gene bcl-2 proteinum. It can supply feasibility bases for the application of thalidomide in uterine cervical cancer. It can offer theoretical support to thalidomide's combined with other chemotherapeutics. It can open up a new way in chems.Methods: 1 Using MTT method determines the inhibition ratio. There are 4 groups: (1) control group. (2) Thalidomide group. Final concentration thalidomide solution is 400mg/L, 200 mg/L, 100 mg/L, 50 mg/L, 25 mg/L, 12.5 mg/L, 6.25 mg/L.(3) combination group. Thd solution as above is combined with 1mg/L cisplatin solution. (4) cisplatin group. Final concentration cisplatin solution is 5 mg/L, 2.5 mg/L, 1 mg/L, 0.5 mg/L.Every density has 3 amb-holes. Determine the absorbance value after 48 hours'cultivation. This experiment repeats 3 times. 3 FCM determines the influence of drugs on cell cycle and apoptosis. 100 mg/L, 50 mg/L, 6.25mg/L thalidomide and 1mg/L cisplatin is adoped. Set groups as above. After 48h, FCM determines the influence of drugs on cell cycle and apoptosis. Repeat 4 times. 4 Immanency to chemical method determines the influence of thalidomide on bcl-2 proteinum. Set 2 group.After 48h, take the cover glasses out and wash then with PBS for 15 minutes. Then , fix them with acetone for 10 minutes and preserve at 4℃. Then operate as the construction on the kit. Repeat 3 times.Result: 1.using MTT method determines the inhibition ratio of drugs. (1) When compare the inhibition ratio of Tha groups with the control group, the differences are significance (P<0.01). There are significant differences among trial groups. The inhibition ratio shows dose-dependent symbiosis. (2) When compare the inhibition ratio of DDP groups with the control group, the differences are significance (P<0.01). There are significant differences among trial groups. The inhibition ratio shows dose-dependent symbiosis. (3) When compare the inhibition ratio of combination groups with the control group, the differences are significance (P<0.01). There are significant differences among trial groups. The inhibition ratio shows dose-dependent symbiosis. Among ,except 6.25mg Thd combined with DDP(62.55 % )p<0.05,there are significant differences among other trial groups. The inhibition ratios are all higher than 2.5mg/L cisplatin (62.11) in group, and it shows dose-dependent symbiosis. 2. Using FCM analyses the influences of the drugs on Hela cells cycle and apoptsis. (1)Thd group. The influence of Thd on the apoptsis ratio shows signigiciant differences when Thd group compares with control group and between two ran groups (p<0.01). And it shows dose-dependent. The influences of Thd on cell cycle show that there are signigiciant differences . (p<0.01) (2) DDP group. The influences of DDP on the apoptsis ratio and on cell cycle are significiantly different when DDP group compares with comtrol group(p<0.01). (3)Combined group. The influences on the apoptsis ratio and on cell cycle are significiantly different when compared with comtrol group and 1mg/L DDP group and Thd groups. And it shows dose-dependent. There are significiant differences between Thd groups and 1mg/L cisplatin group (p<0.01). 3. Using immunocytochemical method determins bcl-2 proteinum. There are differences between Thd groups and the control group (p<0.05,p<0.01) . Conclasion: Thd can inhibit the out-of-body growth of human cervival carcinoma Hela cell lines significantly and influence cell cycle and promote cell apoptsis. These effections are dose-dependent. When Thd combines with cisplatin, synergistic effection and shows dose dependent. Thd can decrease the content of bcl-2 proteinum. This may be one of mechanisms that promotes cell apoptsis.
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