Targeting Senescence: Tianma Granule Inhibits Colorectal Cancer Progression by Modulating miR-29a-5p/P53 Signaling in the Tumor Microenvironment.

LC-MS/MS identified 18 bioactive components in Tianma granule, with network pharmacology analysis highlighting P53 as a core target.

• Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to identify the bioactive components.
• Network pharmacology analysis was conducted to identify key molecular targets.
• P53 was identified as a core target through the network pharmacology approach.
• The identification of P53 as a core target guided subsequent functional investigation of the miR-29a-5p/P53 axis.

TMG suppressed CRC cell proliferation, motility, and invasiveness while promoting apoptosis in vitro.

• Functional assays included CCK-8 (Cell Counting Kit-8) for proliferation assessment.
• Wound-healing and Transwell migration tests were used to assess motility and invasiveness.
• Flow cytometry was used to assess apoptosis.
• Experiments were conducted using doxorubicin (DOX)-induced senescent human umbilical vein endothelial cells (HUVECs) and CRC cell lines.

TMG reduced P53/P21 expression and senescence-associated secretory phenotype (SASP) factors while upregulating miR-29a-5p in senescent HUVECs.

• SASP factors measured included IL-6, IL-8, and CCL20.
• Senescence was induced in HUVECs using doxorubicin (DOX).
• β-galactosidase staining was used to assess cellular senescence.
• TMG treatment reduced both P53/cyclin-dependent kinase inhibitor 1A (P21) and SASP factor levels.
• miR-29a-5p was upregulated in senescent HUVECs following TMG treatment.

miR-29a-5p and P53 exhibit mutual regulatory relationships in the context of senescence-associated signaling.

• Inhibition of miR-29a-5p enhanced senescence and increased P53/P21/SASP levels.
• Silencing of P53 lowered P21 expression and decreased miR-29a-5p levels.
• These results indicate mutual regulation between miR-29a-5p and P53.
• The findings suggest a feedback loop between miR-29a-5p and the P53/P21/SASP pathway.

TMG reduced tumor burden and improved survival in AOM/DSS-induced CRC mice, accompanied by lower P53/P21 expression and restored miR-29a-5p in tissues.

• The in vivo model used was azoxymethane/dextran sulfate sodium (AOM/DSS)-induced CRC in mice.
• TMG treatment resulted in reduced tumor burden in the mouse model.
• Improved survival was observed in TMG-treated AOM/DSS-CRC mice.
• Tissue analysis showed lower P53/P21 levels and restored miR-29a-5p expression in TMG-treated animals.

Chemotherapy-induced senescence-associated tumor microenvironment (S-TME) facilitates colorectal cancer progression.

• Doxorubicin (DOX) was used to induce cellular senescence in HUVECs as a model of chemotherapy-induced senescence.
• The S-TME was characterized by elevated SASP factors including IL-6, IL-8, and CCL20.
• The S-TME was shown to promote CRC tumor growth and metastasis.
• The study investigated how TMG counteracts S-TME-mediated tumor progression.