GRB2 promotes malignant behaviors of breast cancer by modulating the global expression and alternative splicing profiles in SK-BR-3 cells through binding mRNA [fRIP-seq]

The flexible protein GRB2 interacts with HER1~4 on the cell surface and regulates the angiogenesis, differentiation, and proliferation of tumor cells. It is an essential component of the intricate signaling cascade of tyrosine kinase receptors. Meanwhile, GRB2 is an RBP that plays an important role in post-transcriptional regulation in eukaryotes, which affects every stage of mRNA synthesis, modification, splicing, stabilization. Research on breast cancer is limited, but one study found a connection between GRB2 and HER2-positive breast cancer, highlighting the potential of GRB2 as a novel biomarker that stimulates tumor growth. In this research endeavor, we will investigate the impact of GRB2's regulatory mechanism on metastasis in HER2-positive breast cancer. SK-BR3 cells were transfected with a plasmid overexpressing GRB2, and RNA-seq sequencing was performed on the cells before and after transfection. The transcriptome sequencing data before and after GRB2 overexpression were compared, and the differentially expressed genes and alternative splicing events influenced by GRB2 were identified. The fRIP-seg data of GRB2 in SK-BR3 were obtained experimentally, the target mRNA and non-coding RNA bound by GRB2 were compared, the RNA-seg and fRIP-seg data were integrated and analyzed, and the cancer-related pathways and related gene expression/alternative splicing events regulated by GRB2 in SK-BR3 cells were obtained, and the potential functions of GRB2 in HER2+ breast cancer transcription and posttranscription were speculated.