Systems biology pipeline reveals anticancer potential of Curcuma amada in prostate cancer: integrated approach combining network pharmacology, bioinformatics, spatial profiling and experimental validation

Prostate cancer (PCa), a common urinary malignancy, is the leading cause of mortality and morbidity among men worldwide. Curcuma amada extract has demonstrated antitumor properties in preclinical models of various cancers; however, its potential mechanism against PCa remains unclear. Therefore, the current study aimed to investigate the underlying mechanism of C. amada rhizome extract (CARE) in treating PCa through in silico and in vitro approaches. UHPLC-QTOF-HRMS/MS identified 16 phytoconstituents in C. amada, with 15 constituents passing drug-likeness. Public databases identified 1,311 CARE and 473 PCa related targets, with 59 shared targets. Protein–protein interaction analysis revealed P53, CTNNB1, EGFR, AKT1, ESR1, HIF1A, CCND1, PIK3CA, and BCL2 as hub targets. Further, 4-hydroxycinnamic acid, 13-hydroxylabda-8(17),14-dien-18-oic acid, labda-8(17),12-diene-15,16-dial, zederone, zedoarondiol, zerumin A, and caffeic acid were identified as core compounds with high degree values. GO and KEGG analysis-identified targets were primarily associated with apoptosis and the PI3K-AKT signalling pathway. Molecular docking revealed the good binding potential of core compounds with key hub targets. Molecular dynamics showed stable interactions and minimal fluctuations in the complexes throughout the simulation period. Additionally, immunohistochemistry data from the HPA database revealed that AKT1, CTNNB1, EGFR, and PIK3CA proteins showed marked cytoplasmic localization in malignant prostate tissues relative to the adjacent normal epithelium, indicating their potential spatial concentration in cancerous regions. Meanwhile, immune cell deconvolution via the TIMER 2.0 database confirmed that PIK3CA expression showed significant positive correlations with neutrophil and myeloid dendritic cell (r = 0.550 and 0.530, respectively) infiltration, indicating its involvement in establishing the innate immune landscape within the PCa microenvironment. CARE significantly inhibited the proliferation of PC-3 cells, induced apoptosis, and caused G2/M phase arrest. qRT-PCR experiments revealed that CARE suppressed mRNA expression of genes involved in the PI3K-AKT signalling pathway. Thus, this study highlights the therapeutic potential of CARE and elucidates its mechanistic relevance in PCa treatment.