Project description:The excretory-secretory proteins (ESPs) of Trichinella spiralis (T. spiralis) muscle larvae have been confirmed to exert antitumor effects by modulating host immune responses or directly acting on tumor cells. Our research group previously identified that the ESPs of T. spiralis muscle larvae contain multiple proteins potentially related to antitumor activity, among which tropomyosin (TM) has attracted our attention. This study aims to introduce TM from Trichinella spiralis (T. spiralis TM) into breast cancer MCF-7 cells via transfection technology. Furthermore, we investigated the impact of T. spiralis TM on the gene expression profile of MCF-7 cells through transcriptomics. Finally, the study systematically evaluated its regulatory effects on apoptosis, proliferation, and migration. We are the first to comprehensively evaluated the anticancer effects of T. spiralis TM on MCF-7 breast cancer cells. Our findings showed that T. spiralis TM increases apoptosis by activating the mitochondrial-mediated apoptosis pathway and inhibits cell migration and proliferation by downregulating the expression of important hub genes, including PRKACA, CDK3, VPS35, and BCAR1. These findings provide a novel theoretical basis for the application of molecules linked to Trichinella in tumor therapy and support for potential new targets.
Project description:Intro: Spheroid culture especially on MCF-7 cell has been used as in vitro CSC model for anti-CSC drug discovery. The role of miRNAs in the regulation of mRNA specifically looking at the self-renewal capacity and the drug resistance of the spheroid-enriched CSCs models has not been evaluated. Therefore, a comprehensive profiling of the miRNAs will provide an insight into the regulatory mechanisms associated with breast cancer and CSCs. Methods: Tumour spheroid of MCF-7 was generated and characterised for the biological features and CSCs characteristic comparing to the monolayer parental MCF-7 cells. Differential expression of miRNA between the spheroid and parental cells was evaluated using next generation sequencing (NGS). The differentially expressed miRNAs were then subjected to target genes predictions, followed pathway analyses to better understand the mechanism associated with spheroid-enriched CSCs. Results: Spheroids generated from MCF-7 cell line under serum-free condition were found to be enriched with CSCs characteristics. miRNA-NGS analysis revealed 119 differentially expressed miRNAs between the spheroids and parental, with 24 up-regulated and 94 down-regulated. The gene ontology (GO) analysis showed that the predicted genes of the differentially expressed miRNAs were enriched in various biological processes. Pathway analysis revealed that the differentially expressed miRNAs and their target genes were associated with a variety of KEGG pathways, which could explain the self-renewal capability, and the higher drug resistance in spheroids when compared to parental.
Project description:Human breast cancer cell line MCF-7 is usually sensitive to chemotherapy drug BMS-554417, an insulin receptor (IR) and insulin-like growth factor receptor (IGFR) inhibitor. However, through step-wise increase in BMS-554417 doses in culture media, we were able able to screen and select a single MCF-7 clone that is BMS-554417 resistant. It is cross resistant to BMS-536924. This new line of MCF-7 cells was named as MCF-7R4. The transcriptome profiling of both MCF-7 and MCF-7R4 was performed using Affymetrix HG-U133 plus2.0 GeneChip arrays.