The Effect Of Alarelin On Human Endometrial Cancer Xenograft In Nude Mice And The Expression Of P16 And CyclinD1

ObjectiveTo observe the effects of of GnRH-A intervention on the growth of endometrial carcinoma, and reveal the expression of P16 and CyclinD1 ,and also to explore the possible mechanism of GnRH by establishing the model of human endometrial carcinoma xenograft in nude mice .MethodsHuman endometrial carcinoma HEC-1B cells were cultured in vitro to establish animal models of endometrial cancer. Then the successful model nude mice were randomly divided into four groups: control group, low dose group, middle dose group and high dose group. Control group was injected into the muscle with normal saline in 0.2ml. Similarly ,low, medium and high dose group were intramuscularly injected with alarelin of 15,30,60 ug / kg in 0.2ml. Each mice was treated once per day with 6 weeks. After 6 weeks we executed the nude mice , measured the weight ,calculated the inhibition rate, observed the changes of the tumor and detected the expression of P16 and CyclinD1 by immunohistochemical technology (PV-9000) .Results1.There has an irregular mass at their right arm near the maxilla skin, diameter of about 0.4cm ,as the successful model nude mice were inoculated about a week later. The mass has a complete capsule, nodosity, hard and macro- mobile activity partly. The nude mice grow well in a good spirit and normal activities and have no significant changes in diet and urine after inoculation.2.After the treatment was finished , the tumor volume of control group is 697.50±339.93mm3 ; the low dose alarelin group is 552.47±289.33 mm3; the middle dose alarelin group is 457.97±267.51 mm3; the high dose alarelin group , 423.87±268.29 mm3 (P<0.05 ,respectively) . The tumor volume of different doses of alarelin decreases compared with the control group , and the difference was significant(P<0.05 ,respectively). Alarelin inhibited the tumor growth by 20.79%,34.34% and 39.23% in alarelin group of the low ,the middle and the high dose group, and the difference was significant(P<0.05).The difference was significant among the low,the middle and the high dose alarelin group(P<0.05),but the difference was not significant between the middle and the high dose alarelin group(P=0.272).3.The arrangement of the control group neoplastic cells is very labour-intensive and lamellar distribution in the light microscope.These cells have obviously xenotype, abundant cytoplasm ,less interstitial tissue and large nucleus which are very easy to be divided and stained. The arrangement of the Alarelin group is little rarefaction compared with the control group. A large piece of non-structure can be seen in the cells with little minor xenotype. Each group is no significant difference between the general observation. 4.The immunohistochemical results showed that alarelin can increase the expression of P16, but decrease the expression of CyclinD1.The difference between any two groups of all the groups is significant(P<0.05) .Conclusion1.Alarelin with different density has different inhibition to the tumor, and this inhibition has better dose dependent partly.2.Alarelin can change the tumor morphology significantly.3.It may be related to enhance P16 expression and inhibite CyclinD1 expression , to cause to proliferate.