Construction and integration judgment for a mouse model of triple negative breast cancer with liver metastasis of liver qi depression type
Objective To establish a mouse model of liver qi depression triple negative breast cancer with liver metastasis by using chronic unpredictable stress combined with separation.Method In this experiment,6-week-old BALB/c female mice were selected.In the model group,4Tl-luc breast cancer cell line was inoculated in the breast pad for 2 weeks by chronic unpredictable stress(CUS)combined with solitary rearing method and continued to stimulate for 4 weeks.In the same way,6-hydroxydopamine group(6-OHDA group)and Jie Yu Xiao Zheng Granule group were established.Observe and compare the general condition of mice in each group,open field test,sucrose water preference test,serum HP A axis related hormones(CRH,CORT,NE)level,liver HE staining,liver metastasis cell apoptosis level,and the tumor fluo-rescence intensity and the number of liver metastases detected by mouse living imaging.Result There was a statistically significant difference between the control group,model group,Jie Yu Xiao Zheng Granule group,and 6-OHDA group in sugar water preference,activity distance and shuttle frequency scores in the central area of the open field experiment(F=0.34,P＜0.01;F=363.90,P＜0.01;F=0.046,P＜0.001).The preference for sugar water in the model group was significantly lower than the control group,the difference was statistically significant(P＜0.01).The activity distance of the model group was significantly lower than the Jie Yu Xiao zheng Granule group,the difference was statistically signifi-cant(P＜0.01).The shuttle frequency of the model group was significantly lower than the Jie Yu Xiao zheng Granule group,the difference was statistically significant(P＜0.01).The average expression levels of serum CRH,CORT,and NE in the control group,model group,Jie Yu Xiao zheng Granule group,and 6-OHDA group were statistically signifi-cant(F=2 501.00,P＜0.01;F=155.30,P＜0.001;F=149.80,P＜0.01).The expression level of CRH in the mod-el group was 87.71±5.33,which was significantly higher than the Jie Yu Xiao Zheng Granule group,which was 48.87± 3.33,with a statistically significant difference(P＜0.01).The expression level of CORT in the model group was 198.90± 5.64,which was significantly higher than that in the Jie Yu Xiao Zheng Granule group,which was 156.00±5.60,with a statistically significant difference(P＜0.01).The expression level of NE in the model group was 371.00±16.08,which was significantly higher than that in the Jie Yu Xiao Zheng Granule group,which was 262.90±7.98,with a statistically significant difference(P＜0.01).Compared with the Jie Yu Xiao Zheng Granule group,the model group showed a de-crease in the ROI area,a decrease in the number of liver metastases(both P＜0.01).Compared with the model group and the Jie Yu Xiao Zheng Granule group,the apoptosis index of liver metastasis cells of breast cancer in the Jie Yu Xiao Zheng Granule group increased,and the difference was statistically significant(P＜0.01).Conclusion Based on the xeno-graft mouse model of breast cancer,the application of chronic unpredictable stress combined with separation can well sim-ulate triple negative breast cancer with liver metastasis of liver qi depression type,which can provide an experimental car-rier for exploring the essence of stagnation of liver qi depression in triple negative breast cancer with liver metastasis and the pharmacological mechanism of related therapeutic drugs.
triple negative breast cancerliver metastasisliver qi depression syndromechronic unpredictable stressani-mal model
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