| Purpose:The effect of Xihuang pill on the cisplatin sensitivity of the triple-negative breast cancer-bearing nude mouse model established by the cell line MDA-MB-231 was observed.The mechanism of sensitizing cisplatin in triple-negative breast cancer was explored by examining tumor tissue proliferation and apoptotic protein and the expression levels ofβ-catenin,LRP-6 and p-LRP-6 proteins in the classical Wnt/β-catenin signaling pathway.Material and method:(1)Twenty-eight SPF-grade 4-5-week-old female BALB/C nude mice were selected as experimental subjects,and MDA-MB-231 cell suspension was inoculated into the right armpit of nude mice to construct a subcutaneous transplant tumor model of triple-negative breast cancer mice,each 0.2ml.The nude mice that were successful in tumor and did not die were randomly divided into model group(6),Xihuang pill group(6),cisplatin group(6),and cisplatin group(6).Normal saline,Xihuang pill,cisplatin,Xihuang pill combined with cisplatin were used to intervene for 15 days,in which the model group was given 0.2ml/normal saline for gastric lavage,once a day;2mg/kg normal saline intraperitoneal injection,once every 3 days.The Xihuang pill group was given 0.2ml/Xihuang pill suspension for gastric gavage,once a day;2mg/kg normal saline intraperitoneal injection,once every 3days.0.2ml/normal saline gavage was given to the cisplatin group,once a day;2mg/kg cisplatin solution intraperitoneal injection,once every 3 days.The joint group was given0.2ml/Xihuang pill suspension for gastric gavage,once a day;2mg/kg cisplatin solution intraperitoneal injection,once every 3 days.The material is taken every other day after the last dose.(2)After stripping the tumor tissue,the tumor size was compared,the tumor weight was measured and the tumor inhibition rate was calculated.(3)The pathological morphological structure of tumor tissues of each group of mice was observed by hematoxylin-eosin staining(HE).(4)Immunohistochemical staining(IHC)was used to detect the expression of the proliferation-related protein Ki67 in the tumor tissues of each group of mice.(5)Western blot(WB)was used to detect the expression level of proliferation-related protein Cyclin D1 in mouse tumor tissues in each group;The expression level of apoptosis-related protein Bcl-2 in mouse tumor tissues was detected.The expression of Wnt/β-catenin signaling pathway-related proteins β-Catenin,LRP-6 and p-LRP-6 in mouse tumor tissues was detected.(7)SPSS 25 and Image j software were used for data analysis,and P<0.05 was statistically significant,and P<0.01 was statistically significant.Results:1.The tumorigenesis rate of nude mice was 92.8%.During the drug intervention,it was observed that compared with the model group,the tumor growth of Xihuang pill,cisplatin and the combination group was slow,and the drug had an inhibitory growth effect on the transplanted tumor body of the model mouse.After 15 days of administration,the tumor tissue volume in the combination group,cisplatin group and Xihuang pill group was significantly lower than that in the model group(P<0.01),and the tumor tissue volume in the combination group was lower than that in cisplatin group and Xihuang pill group(P<0.01),while there was no significant comparison between Xihuang pill group and cisplatin group(P>0.05).After extraction,the average weight of all group tumor tissues was weighed.Compared with the model group,the tumor weight of Xihuang Pill group,cisplatin group and combination group was significantly lower than that of model group(P<0.01),and compared with cisplatin group and Xihuang Pill group,the tumor weight of the combination group was lower than that of cisplatin group and Xihuang Pill group(P<0.01),while the tumor weight of Xihuang Pill group and cisplatin group was not statistically significant(P>0.05).The tumor suppression rates of Xihuang Pill group,cisplatin group and combination group were 49.7%,56.2% and 72.6%,respectively.2.Observation of the histomorphology of HE staining pathology: the tumor cells in the model group had high density and compactness,and the nuclei were mostly round or oval.Compared with the model group,all drug groups had reduced nucleus density,irregular cell shape,vacuoleization and nuclear fragmentation.Among them,the joint group was more significant than that of Xihuang pill group and cisplatin group.3.After 15 days of administration,the results of Ki67,an important marker related to proliferation activity were observed by immunohistochemical staining(IHC),showing that the expression of Ki67 decreased after treatment in each dosing group and the effect was most significant in the combination group.4.After 15 days of administration,the expression of proliferative protein Cyclin D1,apoptotic protein Bcl-2,and Wnt/β-catenin signaling pathway β-catenin,LRP-6 and p-LRP-6 in each group of tumor tissues was detected by Western blotting method,and the results showed that compared with the model group,the expression of Cyclin D1,Bcl-2,β-catenin,LRP-6,and The expression of p-LRP-6 protein decreased significantly(P<0.01).Compared with Xihuang pill and cisplatin group,the expression of Cyclin D1,Bcl-2,β-catenin,LRP-6 and p-LRP-6 proteins in the combined group was significantly reduced(P<0.01).Conclusion:1.In the nude mouse model of triple-negative breast cancer,Xihuang pill,cisplatin and their combination drugs can inhibit the growth of subcutaneous transplanted tumors to varying degrees.The tumor inhibition effect of Xihuang Pill combined with cisplatin and down-regulation of tumor protein enhancement were better than Xihuang Pill or cisplatin alone,indicating that Xihuang Pill had a synergistic sensitization effect on cisplatin chemotherapy for triple-negative breast cancer.2.The downregulation of tumor proliferative proteins Cyclin D1 and Ki67 and anti-apoptotic protein Bcl-2 by Xihuang pill combined with cisplatin is higher than that of Xihuang pill or cisplatin alone,which further proves that Xihuang pill can increase the sensitivity of triple-negative breast cancer to cisplatin.3.The combination of Xihuang Pill combined with cisplatin significantly reduced the expression of β-catenin,LRP-6 and p-LRP-6 proteins,indicating that the mechanism of Xihuang Pill increasing the sensitivity of triple-negative breast cancer to cisplatin may be related to the inhibition of Wnt/β-catenin signaling pathway. |
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