Animal Experimental Research On The Treatment Of Endometrial Cancer By Blocking The Chemokine CXCL12 Pathway

Objective:1.To improve the nude mice model of endometrial cancer(EC)transplantation in order to shorten the modeling time。2.To explore the effects of blocking the chemokine CXCL12 pathway on the growth of EC transplanted tumor in nude mice and its possible mechanism.Methods:1.PhaseⅠexperiment:High concentration of Ishikawa endometrial cancer cells and Matrigel suspension were prepared at the same volume.The concentration of tumor cells was 7.5*10~7/ml.They were transplanted subcutaneously into the right scapula of nude mice to establish an improved model of EC nude mouse transplantation tumor.The suspension volume of tumor cells was 0.2 ml in each nude mice.2.During the establishment of the tumor model,the growth of nude mice and the growth of transplanted tumors were observed every day.The size of transplanted tumors,including long-diameter a and short-diameter b,was measured by a caliper every 3 days.The volume of transplanted tumors was calculated according to the formula(V=a*b~2/2,mm~3),and the growth curve of transplanted tumors in each experimental group was drawn.Finally,the improved EC tumor model was compared with the tumor model established by the previous research group.3.Tumor tissues were taken for HE staining and slices were observed under optical microscope to see whether the transplanted tumors conformed to the microscopic characteristics of endometrial cancer.4.PhaseⅡexperiment:The model of Ishikawa EC nude mice transplantation tumor was established.Thirty nude mice were randomly divided into 5 experimental groups:AMD3100 treatment group,PD98059 treatment group,CXCR7 neutralizing antibody treatment group,combined drug treatment group and Nacl negative control group.Intraperitoneal injection was used,once every 3 days for 3 weeks,and the growth volume of the transplanted tumor was measured and recorded.5.After 3 weeks of treatment,the nude mice bearing tumor were executed by necking.The transplanted tumors were measured with a caliper and weighed.Calculate the tumor inhibition rate(%)of each experimental group.6.SYBR GREEN real-time fluorescence quantitative RT-PCR was used to detect the expression of vascular endothelial growth factor-C(VEGF-C)in the transplanted tumors,and to verify the therapeutic effect of each drug on endometrial cancer.Results:1.The improved nude mouse model of endometrial cancer was successfully established,which significantly shortened the time of modeling,and the transplanted tumor tissues and cells retained the pathological characteristics of human endometrial cancer.2.Each treatment group could significantly inhibit the growth of EC xenografts in nude mice.The growth of transplanted tumor in AMD3100,PD98059,Anti-CXCR7 and AMD3100+Anti-CXCR7 combined treatment group was relatively slow.After the treatment,thevolume and weight of tumor were significantly smaller than that of Nacl negative control group.There were statistical significance(P<0.01).3.The inhibition rates of AMD3100(55.42±6.32)%,PD98059(60.75±5.75)%,Anti-CXCR7(55.22±5.51)%and AMD3100+Anti-CXCR7(41.71±9.70)%in the four treatment groups compared with those in the saline control group were calculated.There were statistical differences(P<0.01).4.Real-time fluorescence quantitative RT-PCR results showed that the expression of VEGF-C mRAN in transplanted tumors of AMD3100,PD98059 and Anti-CXCR7 treatment groups was significantly lower than that of saline control group(P<0.01).The expression of VEGF-C mRAN in transplanted tumors of AMD3100+Anti-CXCR7 combined treatment group was lower than that of saline control group.There was no statistical significance(P>0.05).Conclusion:The method of establishing EC transplanted tumor model in nude mice by mixing Matrigel and Ishikawa tumor cells is feasible and can shorten the time of modeling,and can be used in the research of EC animal experiments.AMD3100,PD98059 and Anti-CXCR7 can block the chemokine CXCL12 pathway and inhibit the growth of transplanted tumer in nude mice.The mechanism of their effects may be through inhibiting the expression of VEGF-C mRAN in EC cells,which can be tried for further research in the treatment of endometrial cancer.