The Analysis Of Shugan Yishen Recipe’s Effects On Tamoxifen-resistance Breast Cancer Cells

Objective:To study the mechanism of Shugan Yishen recipe reversing drug resistance to tamoxifen in breast cancer cellMethods:1Female SD mice after oophorectomy respectively were given the Shugan Yishen recipe’s decoction (high, medium and low dose group), tamoxifen (TAM) solution, saline (blank group) orally to prepare mice drug scrum.2. MTT assay was used to observe the proliferation inhibition effects of high, medium and low dose of Shugan Yishen recipe group, TAM group, blank group serum on wild-type MCF-7cells and TAM-resistant breast cancer V1CF-7in vitro.3. Transwcl1chamber assay and Wound heal ing scratches assay were used to observe the horizontal and vertical migration Inhibition effects of Shugan Yishen recipe group, TAM group, blank group serums on wild-type MCF-7WT cells and TAM-resistance MCF-7cells in vitro4. After the intervention of Shugan Yishen recipe serum and blank serum on MCF-7CC9in vitro, the Microarray was used to discover the gene expression profiles in differentially groups.5. GO analysis was applied to analyze the main function of the differential expression genes of differential serums according to the Gene Ontology which is the key functional classification of NCBI.6. Go-map analysis was the interaction net of the significant GOs of the differential expression genes, and was made to integrate the relationship between these GOs by outlining the interactions of related GOs and it could summarize the functional interactions of differential expression genes under differential serums.7. Pathway analysis was used to find out the significant pathway of the differential genes of differential group according to KEGG, Biocarta and Reatome.8. The Path-Net was the interaction net of the significant pathways of the differential expression genes, and was built according to the interaction among pathways of the KKGG database to find the interaction among the significant pathways directly and systemically. It could summarize the pathway interaction of differential expression genes of differential groups and found out the reason why certain pathway was activated.9. Gene-gene interaction network was constructed based on the data of differentially expressed genes of different groups. Using Java that allows users to build and analyze molecular networks, network maps were constructed.Results:1.The high-dose group of Shugan Yishen recipe serum and TAM serum inhibited proliferation of MCF-7WT most obviously at48h. The high-dose group of Shugan Yi shen recipe serum also inhibited the proliferation of MCF-7LCC9significantly at48h, but TAM serum could not inhibit the MCF-7FCC9obviously.2The high-dose group of Shugan Yishen recipe serum and TAM serum can inhibit the migration of MCF-7WT and MCF-7LCC9to a certain extent after48hours3After treatments of high-dose group of Shugan Yishen recipe and blank group serums on MCF-7LCC9, Microarray analysis were screened out of1867differentially expressed genes (1.3times), which were up-regulatod1436gene and down-regulated931genes.4. The results of GO-Analysis found133GOs were up-regulated, relating to the mRNA processing, metabolic process, protein transport and so on. While85GOs were down-regulated, relating to response to stimulus, positive regulation of programmed cell death and so on.5. Pathway analysis had found out the significant pathway of the differential genes of differential group. the27up-regulated pathways were RNA degradation, Pyrimidine metabolism. And the21down-regulated pathways were MAPK pathway, VEGF pathway, calcium signaling pathway and so on.Conclusions:1. Shugan Yishen recipe can inhibit the proliferation and migration of MCF-7LCC9.2. Shugan Yishen recipe reverses the tamoxifen-resistance of MCF-7LCC9may be related to the down-regulation of the MAPK pathway, VEGF pathway and AKT3expression.