Rb-driven transcription limits its tumour-suppressive effects in breast cancer

Key Points:

• Various human cancer cell lines, including breast and ovarian cancer lines, were cultured under specific media conditions and authenticated to ensure experimental validity, with mycoplasma contamination routinely checked.
• In vitro drug treatments involved compounds such as abemaciclib, palbociclib, and fulvestrant, with detailed protocols for cell cycle analysis, drug combination assays, and hormone stimulation experiments to assess drug effects on cell proliferation and death.
• In vivo experiments utilized patient-derived xenograft (PDX) models implanted in immunodeficient NSG mice, with randomized treatment groups and blinded downstream analyses to evaluate tumor response to therapies like abemaciclib and palbociclib.
• Advanced molecular techniques including CRISPR–Cas9 gene editing, RNA sequencing, CUT&RUN chromatin profiling, and mass spectrometry were employed to study gene regulation, protein interactions, and chromatin dynamics related to cancer progression and drug response.
• Extensive bioinformatics analyses were conducted on sequencing and proteomics data using established pipelines and software tools for differential expression, motif enrichment, chromatin interaction mapping, and gene set enrichment, ensuring rigorous data interpretation.