Effection And Mechanism Of Alarelin On Subcutaneous Xenograft Growth Of Human Endometrial Carcinoma In Nude Mice

ObjectiveIn the research, we established models of Hunman endometrial carcinoma subcutaneous xenograft by injecting HEC-IB cells subcutaneously. To investigate the effects and its possible molecular mechanisms of different doses of alarelin on human endometrial carcinoma in nude mice of subcutaneous xenograft.MethodsHuman endometrial carcinoma HEC-IB cells were cultured in vitro. We establish a subcutaneous xenograft animal model by injecting HEC-1B cells into the back on the right of the BALB/C nu/nu nude mouse subcutaneously. and removal two nude mouse who is the biggest or the smallest subcutaneous xenograft,then thirty two nude mouse with subcutaneous xenograft were randomly divided into four groups:control group(normal saline), the low dose alarelin group, the middle dose alarelin group, the high dose alarelin group(20,40,80μg/kg, respectively). During the period of treatment, our group members injected the drug by0.2ml every day for four weeks and measured the size and volume of tumor every four days, and growth curves of subcutaneous xenograft tumor were drawed. and the systemic conditions of the mice were observed. Mouse were killed after four weeks of the treatment, removed the xenograft tumor and calculated the volume of tumor. The changes of xenograft morphology were observed by light microscopy with Hematoxylin-easin staining, and the change of Smad2/3and Smad7expressions were estimated by immunohistochemical technologyResults (1) The slope of growth curves of subcutaneous xenograft tumor were reduced and the speed of growth were slowed according to control group,the low dose alarelin group, the middle dose alarelin group, the high dose alarelin group, and the difference was significant(P<0.05, respectively).(2) After four weeks of the treatment, the tumor volume of each group was as follows:control group,1643±252mm3; the low dose alarelin group,1429±299mm3; the middle dose alarelin group,1209±322mm3; the high dose alarelin group,932±238mm3(P<0.05, respectively). and the difference was significant(P<0.05). At the end of the experiment, alarelin inhibited the tumor growth by13.03%、26.42%、43.27%in alarelin group of the low dose.the middle dose and the high dose respectively,and the difference was significant between the groups(P<.05, respectively).(3) The hematoxylin-easin staining results showed that control group filled with tumor cells, and there were obviously xenotype and karyomegaly, karyokinesis. But alarelin group showed significant necrosis morphology changes under a light microscope;(4) The immunohistochemical results showed that alarelin can increase the expression of Smad2/3, but decrease the expression of Smad7. Adding to the doses of alarelin, the expression of Smad2/3increased gradually(P<0.05), but the expression of Smad7decreased gradually (P<0.05). The difference between any two groups of all the groups is significant(P<0.05).Conclusion(1)It is related between The Occurrence and development of Human endometrial carcinoma subcutaneous xenograft and Signal transduction pathways TGF-β1/Smads.(2) Alarelin can significantly suppress tumor growth and to a certain extent dose dependently.(3) Alarelin has an inhibition to the growth of Human endometrial carcinoma subcutaneous xenograft by regulating the expression of Smad2/3and Smad7,which are the key proteins in TGF-β1/Smads signal transduction pathways.