Ethyl Acetate Extract of Antenoron Filiforme Inhibits the Proliferation of Triple Negative Breast Cancer Cell via Suppressing SKP2/P21 Signaling Axis
Purpose: Antenoron Filiforme is a traditionally used herb medicine in China. Our findings indicate that the ethyl acetate extract of Antenoron Filiforme (AF-EAE) has the ability to inhibit the proliferation of triple negative breast cancer cells (TNBC). However, little is known about the underlying mechanism of AF-EAE's action on TNBC. Methods: Here, we performed RNA sequencing on MDA-MB-231 treated by AF-EAE with two concentrations for 24 hours. The sequence reads that passed quality filters were analyzed at the transcript isoform level with hisat2 followed by HTSeq. Results: Using an optimised data analysis workflow, RNA-seq data showed 1,392 differentially expressed genes (DEGs) after low concentration drug treatment and 2,847 DEGs in the high concentration drug treatment group, with a fold change of =1.5 and p-value <0.05. Expression changes in six genes were confirmed by qRT-PCR, demonstrating the high sensitivity of the RNA-seq method. Based on the KEGG pathway enrichment analysis results, cell growth relevant pathways such as "p53 signaling pathway" and "cell cycle" were greatly enriched. In addition, GSEA analysis indicated that DEGs in AF-EAE treated MDA-MB-231 cells were also well enriched in "cell cycle", "DNA replication" and "G2/M cell cycle". Conclusion: Our study indicated that AF-EAE could inhibit the proliferation of  TNBC by inducing cell cycle arrest. Meanwhile, we provide a new potential drug treating breast cancer. Overall design: mRNA profiles of AF-EAE-treated MDA-MB-231 cell line with 2 different concentrations, in duplicate, using Illumina NovaSeq 6000.