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The development of colorectal cancer (CRC) is strongly correlated with the imbalance of various intracellular signal transduction cascades, including protein kinase B (AKT), mitogen-activated protein kinase 1 (MAPK), signal transducer and activator of transcription 3 (STAT3), as well as crosstalk between these signaling networks.At present, antitumor agents are often single-targeted and thus are not always therapeutically effective.Moreover, long-term use of these antitumor agents often generates drug resistance and potential side-effects.These problems highlight the urgent need for the development of novel and more effective anticancer drugs.Hedyotis Diffusa Willd (HDW) has been used as a major component in traditional Chinese medicine for the clinical treatment of colorectal cancer, with few adverse effects.However, the molecular mechanisms which form the basis for its anti-cancer activity still require further elucidation.In the present study, using xenograft models and various different human CRC cell lines, we evaluated the efficacy of the ethanol extract of HDW (EEHDW) against tumor growth and investigated its underlying molecular mechanisms of action.We demonstrated that EEHDW robustly inhibits cancer growth both in vivo and in vitro.Furthermore, EEHDW suppressed the activation of several CRC-related signaling pathways and can regulate the expression of various inflammatory and angiogenic factors.This ultimately led to the induction of apoptosis and inhibition of cellular proliferation, as well as tumor angiogenesis.Our study demonstrates that EEHDW possesses extensive anticancer activity due to its ability to affect multiple intracellular targets, suggesting that HDW is a novel multipotent therapeutic agent for colorectal cancer treatment.