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Based on network pharmacology analysis, this study was to uncover the anti-colorectal cancer (CRC) targets and molecular mechanisms exerted by puerarin. Pathological genes of CRC and therapeutic genes of puerarin were collected through well-established databases. The crucial targets of puerarin against CRC were further used for function and pathway enrichment assays to elucidate the biological processes and signaling pathways, followed by experiment-based verification. In network data, the most significant targets of tyrosyl-DNA phosphodiesterase-1 (TDP1), aldehyde dehydrogenase 1 family member A-1 (ALDH1A1), muscleblind like splicing regulator 1 (MBNL1), aldehyde dehydrogenase-2 (ALDH2), and nicotinamide adenine dinucleotide (HPGD) were screened and defined in anti-CRC effects exerted by puerarin. In further enrichment assays, the functional processes of puerarin against CRC were associated with energy pathways, metabolism, cell communication, signal transduction, aldehyde metabolism, DNA repair. Meanwhile, key ten signaling pathways from bioinformatic findings were ascertained respectively. As revealed in human data, CRC patients showed up-regulated expressions of endogenous TDP1, ALDH1A1, accompanied with visible hematoxylin-eosin (HE) and Ki-67 stains and elevated blood tumor marker expressions. In further study in vitro, puerarin-treated human CRC cells resulted in inhibited cell growth, increased cell apoptosis in a dose-dependent way. Further, down-regulated expressions of TDP1, ALDH1A1 and proliferating cell nuclear antigen (PCNA) were detected in puerarin-treated CRC cells. In conclusion, the molecular network data manifest the biotargets and signaling pathways of puerarin against CRC, followed by verification of both human and cell line studies. Furthermore, the pharmacological molecules of TDP1, ALDH1A1 seem to be the possible targets for managing CRC.
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