Study the anti-tumor activity and mechanism of action of chalcone natural products. The antitumor activity of 21 chalcone natural products (1-21) was evaluated by MTT assay. The most effective natural products and their concentrations were chosen to intervene in human in situ pancreatic adenocarcinoma BXPC3 cells, human colorectal adenocarcinoma SW620 cells, human prostate cancer DU145 cells, human thyroid cancer TPC-1 cells and human liver cancer HUH-7 cells. Difference analysis and enrichment analysis were performed using transcriptome sequencing technology. The expression of signal pathway proteins was detected by Western blot (WB) method. The results showed that compounds 1-21 had good anti-tumor activities against BXPC3 cells, among which butein (21) had the strongest inhibitory effect with an IC₅₀ of 4.91 ± 0.93 μmol/L. licochalcone A (1), flavokawain B (6) and xanthohumol (9) showed good anti-tumor activity against BXPC3, SW620, DU145, TPC-1 and HUH-7 cells. After the intervention of BXPC3, SW620 and DU145 cells with compounds 1 and 6, the total number of differentially expressed genes in the three groups was 10 and 3, respectively. KEGG enrichment analysis results showed that differentially expressed genes were mainly concentrated in interleukin-17 (IL-17) signaling pathway. WB results showed that compared with the control group, the protein expression of IL-6, tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-9 (MMP-9) was reduced in DU145 cells treated with compounds 1, 6 and 9. Therefore, chalcone natural products may exert anti-tumor effects by regulating the IL-17 signaling pathway and inhibiting IL-6, TNF-α and MMP-9.