5,6,7,4'-Tetramethoxyflavone, a Dietary Polymethoxyflavone, Exerts Antitumor Effect on HeLa Xenografts in Mice

5,6,7,4'-Tetramethoxyflavone (TMF), a naturally occurring polymethoxyflavone (PMF) abundant in Citrus species, has demonstrated potent antitumor activity against HeLa cells in vitro. To extend these findings, this study systematically investigated its therapeutic efficacy and safety profile in a HeLa tumor xenograft model. Here, we first found that TMF induced apoptosis in HeLa cancer cells both in vitro and in vivo. Proteomics analysis identified 19 differentially expressed proteins (DEPs) from HeLa cancer cells after TMF treatment, including downregulation of HSP60, sTNF-R1, JNK, TAK1 (S412), TBK1 (S172), ZAP70 (Y292), ATF2, c-Fos, c-JUN, Smad1, Smad5, and Stat6 (Tyr64), alongside upregulation of sTNF-R2, AKT, GSK3b, MKK3, MKK6, MSK2, and P38. Transcriptomics analysis further uncovered that there were 261 differentially expressed genes (DEGs). Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment for the 19 DEPs and 261 DEGs revealed that the underlying mechanisms of TMF against HeLa tumor pointed primarily to MAPK, TNF, VEGF, Ras, and FoxO signaling pathways. Notably, histopathological evaluation revealed no observable tissue damage in major organs (liver, kidney, lung, heart, spleen) following TMF administration. In addition, the results of biochemical indexes further verified that the overall changes in plasma ALT, AST, TBIL, DBIL, TRIG, ALP, LDH, GGT, CREA, UA, UREA, CK, HBD, and CHOL were significantly smaller in the TMF-treated groups than in the DDP-treated groups. These findings collectively position TMF as a promising therapeutic candidate combining robust antitumor efficacy with favorable safety characteristics for the treatment of cervical cancer.