| Objective: In this study, preliminary studies on the basis of the test through in vivo observation of the effects of AG490 combined DDP chemotherapy drugs on tumor growth in nude mice. Discussing AG490 by blocking STAT3 pathway on the effects of tumor and the relationship between the Caspase-3 expression and STAT3 pathway. With a view to looking for better ways of treatment.Methods: We established the cervical cancer model in nude mice with Hela cell and gave the control group, DDP, AG490, AG490 +DDP by intraperitoneal injection. The weight of nude mice and the tumor size was measured. The tumor weight and inhibition rate were evaluated. When the nude mice were sacrificed after 2 weeks treatment. Cell apoptosis was detected by flow cytometry (FCM). The necrotic degree and range were observed by the microscope. Statistical software SPSS10.0 deal with all data.Results:1. The effect of the drug to nude mice tumor volume : Tumor volume in DDP group, AG490 group and combination group was less than the control group, there are significant differences(P<0.001). In the treatment of the first 7 days. Compared with the combination group, there are significant differences(P<0.001) in DDP group and AG490 group(P<0.05,P<0.001). There are significant differences between DDP group and AG490 group at inhibiting tumor growth(P<0.05).2,The effect of the drug to nude mice body weight: During the drug treatment of cervical cancer nude mice growth , there have a change in body weight compared with the control group, there are significant differences in DDP group and combination group (P<0.001). There are no significant differences between AG490 group and the control group (P>0.05). Nude mice are generally good to eat and no significant changes in body weight in AG490 group. However, in DDP group, at the beginning of the treatment on the 7th day the nude mice has become more and more severe weight loss, malnutrition, there are significant differences compared with combination group in weight change (P<0.05). One nude mice death.3. Each group compared with inhibition rate DDP group, AG490 group, DDP group + AG490 group of nude tumor weight inhibition rate was respectively 51.2%, 33.6% and 59.2%,which have a significant reduction compare with control group(P<0.05).Combination group have a significant reduction in tumor weight compared with the DDP group, AG490 group(P<0.05). There are significant differences between DDP group and the AG490 group (P<0.05)4. Flow cytometry: DDP group, AG490 group and the combination group marked the peak of apoptosis in G1 phase. The apoptosis rate of the three groups were 52.1±1.3%, 39.5±0.4% and 66.7±1.8%, there are significant differences compared with the control group 10.2±2.4% (P<0.05).5. Pathological observation: Seeing with naked-eye that tumor was nodular, tumor tissue rich in blood supply, no surrounding tissue and skin infiltration, the boundaries are clear and easy separation, the cut surface like fish. Tumor cells were arranged in nests, films, cord-like and were large, obviously nucleolus, deep-dyeing in light microscope to observe. Pathological mitotic phase are easy to see. In the tumor tissue, blood vessels are expansion congestive and a small amount of lymphocytes, neutrophil infiltration. There have a small number of fibers and blood vessels in between the tumor nest. Small focal necrosis and coagulation necrosis can be seen in the tumor tissue. After the treatment, tumor necrosis occurred in varying degree, the results showed that the degrees of necrosis were varied in each group. The average rate of tumor necrosis: control group: 21.8±9.0%; DDP group: 40.8±7.3%; AG490 group: 39.4±11.3%; the combination group: 61.9±9.6%. There are significant differences compared with the control group (P<0.05). There are significant differences between the combination group and AG490 group,DDP group(P<0.05).6. The results of immunohistochemistry: P-stat3 positive expression of the cancer cells are mainly located in the cytoplasm, deep-dyeing and scattered. Caspase-3 positive expression of the cancer cells are also located in the cytoplasm, scattered or focal distribution. P-stat3 positive rate was 92.5% in control group,64% in DDP group,55% in AG490 group and 40.9% in combination group . Caspase-3 positive rate was 30.4% in control group,60% in DDP group,55.6% in AG490 group and 92.3% in combination group. There are no significant differences between P-stat3 protein and caspase-3 protein in AG490 group. (r=-0.074,P>0.05).Conclusion:1. There is inhibition of AG490 on human cervical cancer Hela cells growth of nude mice , it can increase the DDP sensitivity to cervical cancer drug and reduce the DDP toxicity. AG490 therapy combined with DDP therapy significantly enhance the effect of inhibition.2.AG490 can induce apoptosis of transplanted tumor, combined application of cisplatin enhanced its effect.3.AG490 therapy combined with DDP therapy can reduce the expression of P-stat3 at nude mice, indirectly increase the expression of caspase-3, however, no direct effects.
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