Synergistic targeting of ITGA5/CCN1 by Sparganii Rhizoma and Curcumae Rhizoma suppresses breast cancer metastasis via angiogenesis and adhesion.

BACKGROUND

Despite significant progress in medical research, breast cancer remains the second most prevalent and lethal malignant tumor among women, with metastasis being a major contributor to patient mortality. Sparganii Rhizoma-Curcumae Rhizoma (SR-CR), a well-recognized traditional Chinese medicine (TCM) formulation, exhibits potent anti-tumor effects. Its primary bioactive components, Flavonoids of Sparganii Rhizoma (FSR) and Oil of Curcumae Rhizoma (OCR), have been shown to possess a broad spectrum of pharmacological activities, including anti-tumor, cardiovascular, antithrombotic, blood rheology-enhancing, and anti-breast hyperplasia properties. The synergistic use of Sparganii Rhizoma and Curcumae Rhizoma is a common practice in TCM for cancer treatment. However, the combined effect of FSR and OCR on breast cancer remains insufficiently investigated.

PURPOSE: This study aims to evaluate the therapeutic potential of FSR + OCR and elucidate its pharmacodynamic mechanisms.

STUDY

DESIGN

The therapeutic efficacy of FSR + OCR was evaluated using both a zebrafish xenograft tumor model and a mouse spontaneous breast cancer metastasis model.

METHODS

Fluorescently labeled MDA-MB-231 cells were microinjected into the yolk sac of 48-hour-old zebrafish embryos, and the subintestinal vessels, as well as tumor cell metastasis, were examined using a fluorescent microscope. In the murine model, a 4T1 cell suspension was injected into the left fourth mammary fat pad in Balb/c mice (female). The impact of FSR + OCR on tumor metastasis was assessed via flow cytometry, hematoxylin-eosin staining, immunohistochemistry, and immunofluorescence. Potential drug targets were identified through transcriptome sequencing. The mechanisms underlying the effects of FSR + OCR on tumor metastasis were further confirmed through CCK-8 assays, Western blot analysis, qRT-PCR, tube formation assays, and transendothelial migration assays.

RESULTS

Administration of FSR + OCR at varying doses did not inhibit tumor growth significantly; however, medium and high doses effectively suppressed tumor angiogenesis and metastasis. Transcriptome sequencing revealed potential drug targets through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, including integrin alpha 5 (ITGA5) and cellular communication network factor 1 (CCN1), which were further validated through GO and KEGG analyses.

CONCLUSION

In conclusion, the combination of FSR + OCR inhibits breast cancer metastasis by disrupting ITGA5-mediated tumor neovascularization and attenuating CCN1-mediated enhancement of vascular endothelial adhesion. This study combines "blood stasis" theory in TCM with vascular biology research, and deciphers the synergistic anti-metastasis mechanism of Sparganii Rhizoma-Curcumae Rhizoma on the molecular level.