Abstract: This study aimed to investigate the inhibitory effect and mechanism of lotus seedpod polysaccharides (LSP) on cervical cancer HeLa cells using commercial kits, flow cytometry, cell scratch wound healing assay, and Transwell cell migration and invasion assays. Transcriptomics, proteomics, and Western blot were integrated to explore the potential mechanism by which LSP inhibit HeLa cells. The results indicated that LSP significantly reduced HeLa cell viability, affected cell morphology, induced apoptosis and mitochondrial membrane potential collapse, increased reactive oxygen species (ROS) production, and inhibited cell migration and invasion. Transcriptomic analysis revealed that 120 μg/mL LSP significantly regulated the expression of 12 347 genes (5 550 upregulated, 6 797 downregulated) in HeLa cells, altering biological processes, molecular functions, and cellular components, and modulating metabolic pathways, oxidative phosphorylation, chemical carcinogenesis-ROS, and the cell cycle. Proteomic analysis showed that 120 μg/mL LSP significantly regulated the expression of 1 143 proteins (379 upregulated, 764 downregulated) in HeLa cells, altering biological processes, molecular functions, and cellular components, and modulating several pathways including the pentose phosphate pathway, purine metabolism, glycolysis/gluconeogenesis, and nucleotide metabolism. Integrated transcriptomic and proteomic analysis revealed 1 041 overlapping differentially expressed proteins and genes, which were significantly enriched in glycolysis/gluconeogenesis, amino sugar and nucleotide sugar metabolism, the pentose phosphate pathway, glutathione metabolism. Further investigation focusing on the glycolysis/gluconeogenesis pathway revealed that LSP significantly decreased adenosine triphosphate (ATP) content, glucose consumption, and lactate production in HeLa cells, inhibited the activities of hexokinase (HK), phosphofructokinase (PFK), and pyruvate kinase (PK), and downregulated the protein expression levels of glycolytic enzymes, including glucose-6-phosphate isomerase (GPI), phosphofructokinase platelet-type (PFKP), phosphofructokinase muscle-type (PFKM), fructose-bisphosphate aldolase A (ALDOA), triosephosphate isomerase 1 (TPI1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase 1 (PGK1), phosphoglycerate mutase 1 (PGAM1), enolase 1 (ENO1), enolase 2 (ENO2), enolase 3 (ENO3), pyruvate kinase M (PKM), lactate dehydrogenase A (LDHA), and lactate dehydrogenase B (LDHB). In conclusion, LSP have a significant inhibitory effect on HeLa cells, which may be related to their significant suppression of the glycolysis pathway. These findings provide a theoretical basis for the functional development and high-value utilization of LSP for the prevention and treatment of cervical cancer.

Key words: lotus seedpod polysaccharides; cervical cancer HeLa cells; transcriptomics; proteomics; mechanism; glycolysis pathway