| AIM:Increasing evidence has shown that Chinese herbal medicine(CHM)has promising therapeutic effects in colorectal cancer(CRC);however,the active ingredients and potential targets remain unclear.In this study,we aimed to investigate the relative molecular targets of the Chinese herbs that have been found effective in treating metastatic CRC(mCRC)based on clinical data and network pharmacology.MOTHODS:1.Collect clinical cases and follow up :After analysis of the collected data and telephone follow-ups,the medical records of 222 patients diagnosed with mCRC between November 2007 and April 2012 were retrospectively reviewed.2.Kaplan–Meier curves and multivariate Cox regression analyses were used to compare the differences in survival time.P < 0.05 was considered statistically significant.3.Spearman's bivariate correlation analysis was used to determine the correlation between the herbs and survival time and identify the most effective components in the formula.P < 0.01 was considered statistically significant.And a network pharmacological approach was developed to elucidate the potential therapeutic mechanisms of CHM.4.Experimental validation : Colorectal carcinoma cell line HT29 was used.The anti-colorectal cancer effects of aqueous extract of 18 herbs were tested.Cell proliferation and cell migration were evaluated using typanblue staining assay and wound healing assay,respectively.The western blotting was adopted to prove the predicted targets.RESULTS:1.A total of 222 patients with mCRC were included in this study.Among them,78 patients received CHM treatment,and 144 patients received non-CHM treatment.2.Kaplan–Meier and Cox regression analyses showed that CHM was an independent protective factor in patients with mCRC.The hazard ratio was 0.103(95 % confidence interval = 0.064–0.164;P < 0.001).Patients in the CHM groupexhibited a longer median survival time(40 months)compared to those in the non-CHM group(12 months;P < 0.001).3.Eighteen out of 295 herbs significantly correlated with favorable survival outcomes(P < 0.05).4.Targets prediction: RXRA,TOP2,TNF,PPAR?,AKT1,SRC,ERBB2,VEGFR,and VEGFA had the most of the direct interactions with these herbs,suggesting that these proteins might play important roles in the treatment of CRC.5.Our results revealed that aqueous of 18 herbs could obviously inhibit cell proliferation and cell migration(12h)in dose of 100 ug/mL,200 ug/mL,300 ug/mL and 400 ug/mL in vitro.Furthermore,18 herbs significantly inhibited the expression of ERBB2,AKT1,VEGFA and VEGFR in dose of 300 ug/mL and 400 ug/mL.CONCLUSION:In the present study,we showed that CHM was associated with survival benefit in patients with mCRC;in addition,correlation analysis identified 18 herbs with positive effects on survival.Moreover,we used a network pharmacological approach to investigate the underlying mechanisms,which showed that 18 herbs in CHM might exhibit anti-CRC activity mainly via 1)inhibition of the proliferative activity of ERBB2,PPAR?,and RXR;2)suppression of angiogenesis by inhibiting VEGFR and VEGFA expression;3)inhibition of the PI3K/AKT1 signaling pathway directly through SRC and AKT1;and 4)Reduction of TNF-induced inflammation. |
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