The Inhibitive Effects Of Purified Realgar And White Arsenic On Cervical Cancer And Its Influence On Apoptosis

Objective1.By transplanting the Hela cell to the nude mice,we study the tumor suppressive effect of purified realgar and white arsenic to cervical cancer and explore the side effects and toxicities of them to the mice.2.To explore the mechanism of inhibiting tumor growth in purified realgar and white arsenic by analyzing the apoptosis and cell cycle.3.To investigate the relation between the apoptosis and the protein expression of Fas,Caspase-3 and Survivin.Methods1.Hela cells were cultured at 37℃in a 5 mL/L CO₂ incubator.Seventy mice received subcutaneous injection with cell suspension containing 2×10⁷ cells.When tumors of about 0.5cm diameter were detected,drug administration was started.The animals were randomly divided into seven groups including model group,carboplatin group,white realgar group,low dose purified realgar group,middle dose purified realgar group,high dose purified realgar group and purified realgar combined with white arsenic group.There are 10 animals in each group.After 15d of treatment,the mice were sacrificed and the tumor masses were removed.After the weight of tumor masses was measured,some were fixed in 10%paraformaldehyde,and some were frozen.The tissues of liver,kindy,spleen,stomach,large intestine and small intestine were stained with HE and analyzed for the side effects and toxicities of the drugs.2.Tumor tissues were homogenated,washed with PBS and then filtered.The tumor cells were mixed with PBS to the concentration of 1×10⁶ cells/ml.After adding annexin V-FITC and PI,the tumor cells were analyzed by flow cytometer for the apoptosis and cell cycle.3.Fas protein,Caspase-3 protein and surviving protein were detected with Immunohistochemical SP method.Results1.The weights of the tumors in carboplatin group,white realgar group,middle dose purified realgar group,high dose purified realgar group and purified realgar combined with white arsenic group are more than in the model group(P<0.05).In the all treatment groups,there was no differences in weights of the tumors(P>0.05).The suppressive rates to tumor in carboplatin group,white realgar group,middle dose purified realgar group, high dose purified realgar group and purified realgar combined with white arsenic group was 34.3%,35.8%,37.0%,33.1%,37.8%respectively.These suggest that purified realgar and white arsenic can inhibit the proliferation of cervical cancer,and purified realgar and white arsenic have no toxicity to liver,lung,kidney,spleen,stomach,large intestine or small intestine.2.The apoptotic rate of cervical cancer cells treated with purified realgar and white arsenic were significantly higher than that in the model group (P<0.05).3.The cervical cancer cells in G₀/G₁ phase in the treatment groups were much more than in the model group(P<0.05).This suggests purified realgar and white arsenic arrest the cervical cancer cells in G₀/G₁ phase.4.The expressions of Fas protein in the treatment groups were much higher than in the model group(P<0.05).The expressions of Caspase-3 protein in the treatment groups were much higher than in the model group(P<0.05).The expressions of Survivin protein in the treatment groups were much lower than in the model group(P<0.05).Conclusion1.As the carboplatin,purified realgar and white arsenic can inhibit the proliferation of cervical cancer.If realgar is given to the nude mice by filling stomach combining with local injecting method,the tumor supprcssive effect is the best.The suppressive effect of purified realgar is not in a dose-dependent manner,which may be related with its characteristic of no dissolving to water. 2.Purified realgarand white arsenic induced apoptosis of cervical cancer. Inducing apoptosis may be one of the mechanisms of the tumor suppressive effect.3.Purified realgar and white arsenic arrest the cervical cancer cells in G₀/G₁ phase.They delay the proliferation of cervical cancer,and make the apoptosis easy.4.Purified realgar and white arsenic increase protein expression of Fas and Caspase-3,and decease Survivin.UP-regulating of Fas and Caspase-3 and down-regulating of Survivin may play an important role in the apoptosis in cervical cancer cells.