Study On Lungs Metastasis From Breast Cancer Intervened By Yanghe Decoction Based On PHD2/TGF-?/CAF Path

?Objective?In this study,the path of PHD2/TGF-?1/CAF was choosed as an important role in the process of breast cancer metastasis.To validation the effect of Yanghe Decoction intervene lung metastases from breast cancer By the model of nude mice.Then observe the influence of different dosages on the expression of PHD2,HIF-1,TGF-1 and CAF in tumor microenvironment.To explor the mechanism of Yanghe Decoction in therapy of lung metastases from breast cancer though improving the tumor microenvironment.?Methods?1.To build an animal model of lung metastases from breast cancer in nude mice.Inoculate with 0.2 ml of MDA-MB-231 cell suspension(concentration 5×10~7/ml)on the second breast fat pad in the right chest of the nude mice.Observe the proliferation of tumor cells.It is successful when the proliferation is visible to the naked eyes.Evaluate the model was build successfully by observe the histological morphology in pathological sections.2.To study the Effect of Yanghe Decoction intervene tumor formation and metastasis on nude mice.30 nude mice were randomly divided into 5 groups which are model group,control group,Yanghe Decoction high dose group,Yanghe Decoction middle dose group,Yanghe Decoction low dose group.And 6 rats in each group.6 healthy rats were selected as nomal control group.Decoction intervene for21 days.Anatomy to take tumor and lungs.Observation of lung metastases in nude mice by HE.Draw curves of nude weight and tumor growth.Calculate the tumor inhibition rate.3.To study the relationship between Yanghe Decoction treat breast cancer and Yanghe Decoction adjust the expression PHD2,HIF-1?,TGF-?1 and CAF.Detection the expression PHD2,HIF-1?and CAF of tumor by IHC while TGF-?1 by RT-PCR.4.To study the relationship between Yanghe Decoction treat lung metastases from breast cancer and Yanghe Decoction adjust the expression PHD2,HIF-1?,TGF-?1 and CAF.Detection the expression PHD2,HIF-1?and CAF of lungs by IHC while TGF-?1 by RT-PCR.?Results?1.It is successful that building animal model with a rate of 100%.Breast cancer orthotopic tumors are located in the right side of the second nipple of the nude mice.And the border is clear,the shape is rule,palpation activity is acceptable,and the capsule is complete.Only part of has adhesion.The situ tumor looks like fish color,tough quality,has gray necrotic tissue in center after cuted.In situ tumor tissue and lung tissue can see the tumor by HE.2.In situ tumor tissue can see the tumor of each group by HE while in lungs can see the tumor except the nomal group and the contral group.Draw curves of tumor growth.The tumor inhibition rate are contral group(51.44%),Yanghe Decoction high dose group(18.16%),Yanghe decoction middle dose group(26.37%),Yanghe decoction low dose group(9.22%).3.After treatment,the expression of the various indicators of the situ tumors were as follows.The expression of PHD2 in the control group and Yanghe Decoction middle dose group were significantly higher than that in the model group,all had statistical significance(control group P<0.01;Yanghe Decoction middle dose group P<0.05).The expression of HIF-1?in the control group and Yanghe Decoction middle dose group were lower than that in the model group,all had statistical significance(control group P<0.01;Yanghe Decoction middle dose group P<0.05).The expression of?-SMA in the control group was lower than that in the model group,the difference was statistically significant(P<0.01).The expression of TGF-?1mRNA in the control group was lower than that in the model group,the difference was statistically significant(P<0.01).4.After treatment,the expression of the various indicators of the lungs were as follows.The expression of PHD2 in the nomal group was higher than that in the other groups,the difference was statistically significant(P<0.01).The expression of PHD2in the model group was higher than that in the other groups except the nomal group,all had statistical significance(Yanghe Decoction low dose group P<0.05;the others P<0.01).The expression of HIF-1?in the nomal group was lower than that in the other groups,all had statistical significance(the model group P<0.01;the others P<0.05).The expression of HIF-1?in the model group was higher than that in the other groups except the nomal group,the control group had statistical significance(P<0.05).The expression of?-SMA in the nomal group was higher than that in the other groups,the difference was statistically significant(P<0.01).The expression of?-SMA in the model group was higher than that in the other groups except the nomal group,the control group had statistical significance(P<0.01),the Yanghe Decoction middle dose group had statistical significance(P<0.05).The expression of TGF-?1mRNA in the nomal group was lower than that in the other groups,the difference was statistically significant(P<0.01).The expression of TGF-?1mRNA in the model group was higher than that in the other groups except the nomal group,the control group and the Yanghe Decoction middle dose group had statistical significance(P<0.05).?Conclusions?1.Yanghe decoction inhibit the growth of breast cancer in situ tumor by increasing the expression of PHD2 while reducing the expression of HIF-1?.It intervene hypoxic microenvironment to achieve those effects.The expression of TGF-?1 and?-SMA(CAF)in each group showed that Yanghe Decoction did not regulate TGF-?1 and?-SMA to treat breast cancer in situ tumor.2.Yanghe decoction inhibit the infiltration of lung metastases from breast cancer by reducing TGF-?1,?-SMA(CAF)in lung tissue of nude mice to intervene inflammatory microenvironment.The expression of HIF-1?in each group showed that Yanghe Decoction did not regulate HIF-1?to treat lung metastases from breast cancer.