Search

Objective We aimed to investigate the mechanisms underlying the effects of Huangqi and Ezhu combined with 5-fluorouracil (5-FU) in the treatment of colon cancer. Methods One male BALB/c mouse was used to generate the orthotopic tumor, and orthotopic tumor tissue was transplanted into 80 male BALB/c mice to establish the CT26. WT mouse model. These 80 mice were randomly divided into (i) the model group, (ii) the 5-FU group (25 mg/kg), (iii) the high-dose huangqi plus ezhu group (12 g/kg), (iv) the mid-dose huangqi plus ezhu group (6 g/kg), (v) the low-dose huangqi plus ezhu group (3 g/kg), (vi) the high-dose combination therapy group (12 g/kg + 25 mg/kg), (vii) the mid-dose combination therapy group (6 g/kg + 25 mg/kg), and (viii) the low-dose combination therapy group (3g/kg +25 mg/kg) (n = 10 mice per group). Treatment lasted for 3 weeks. The tumor inhibition rate was calculated by collecting tumors after the mice were killed. Chemokin (C-X-C) motif ligand 10 (CXCL10), Chemokin (C-X-C) motif receptor 3 (CXCR3), Chemokin (C-C) motif ligand 3 (CCL3), and Chemokin (C-C) motif receptor 5 (CCR5) mRNA and protein expression levels in tumor tissue, lymphoid tissue, and liver tissue were measured by RT-PCR and western blot, respectively. Results Compared with model group, the tumor inhibition rate in each treatment group was increased (P < 0.01). Compared with 5-FU group, the tumor inhibition rate of high-dose, mid-dose, low-dose huangqi plus ezhu, groups and high-dose, mid-dose, low-dose combination therapy group was decreased (P < 0. 01). Compared with the high-dose huangqi plus ezhu group, the tumor inhibition rate of high-dose combination therapy group was increased (P < 0. 01), and low-dose huangqi plus ezhu groups were decreased (P < 0. 01). Compared with mid-dose huangqi plus ezhu group, the tumor inhibition rate of low-dose huangqi plus ezhu group was decreased (P < 0. 01), and the mid-dose combination therapy group group was increased (P < 0. 01). Compared with the high-dose combination therapy group, the tumor inhibition rate of mid-dose combination therapy group was increased (P < 0. 01). Compared with mid-dose combination therapy group, the tumor inhibition rate of low-dose combination therapy group was decreased (P <0. 01). Compared with the low-dose combination therapy group, the tumor inhibition rate of low-dose huangqi plus ezhu group was decreased(P <0. 01). Compared with the model group, mRNA and protein levels of CXCL10, CXCR3, CCL3, and CCR5 were lower in each treatment group (P <0. 01). Compared with the 5-FU group, mRNA and protein levels of CXCL10, CXCR3, CCL3, and CCR5 were higher in the mid- and low-dose huangqi plus ezhu groups (P <0. 05, P <0. 01) and lower in the combination therapy groups (P <0. 01). Compared with the 5-FU group, the mRNA and protein levels of CCL3 in orthotopic tumor tissue, lymph node tissue, and liver tissue and the protein levels of CCR5 in the liver tissue of the high-dose huangqi plus ezhu group were increased (P <0. 01), and the mRNA levels of CXCL10 and CXCR3 in the high-dose huangqi plus ezhu group were increased (P <0. 05, P <0. 01). Compared with the high-dose huangqi plus ezhu group, mRNA and the protein levels of chemokines and receptors were decreased in the high-dose combination therapy group (P < 0. 05), the protein levels of chemokines and receptors in orthotopic tumor tissue and lymph node tissue were increased in the mid-dose and low-dose huangqi plus ezhu groups (P < 0. 05), the protein levels of CXCL10, CXCR3, and CCR5 in liver tissue were increased in the mid-dose and low-dose huangqi plus ezhu groups (P < 0. 05), the mRNA levels of chemokines and receptors were increased in the low-dose huangqi plus ezhu group(P <0. 05), and the mRNA levels of CXCR3, CCL3, and CCR5 were increased in the mid-dose huangqi plus ezhu group (P <0. 05). Compared with the mid-dose huangqi plus ezhu group, the mRNA and protein levels of chemokines and receptors were decreased in the mid-dose combination therapy group (P < 0. 05), the protein levels of chemokines and receptors in orthotopic tumor tissue were increased in the low-dose huangqi plus ezhu group (P < 0. 05), the protein levels of CXCL10 and CCL3 in lymph node tissue were increased (P < 0. 05), the protein levels of CXCL10 and CCR5 in liver tissue were increased (P <0. 05), and the mRNA levels of CXCL10 and CCL3 were increased in the low-dose huangqi plus ezhu group (P <0. 05). Compared with the high-dose combination therapy group, the mRNA and protein levels of chemokines and receptors were increased in the low-dose combination therapy group (P < 0. 05), the protein levels of CCL3 ad CXCL10 in orthotopic tumor tissue were increased in the mid-dose combination therapy group (P < 0. 05), the protein levels of CCR5, CXCL10, and CXCR3 in lymph node tissue were increased in the mid-dose combination therapy group(P <0. 05), the protein levels of CXCL10 and CXCR3 in liver tissue were increased in the mid-dose combination therapy group (P <0. 05), and the mRNA levels of CXCR3 were increased in the mid-dose combination therapy group (P <0. 05). Compared with the mid-dose combination therapy group, the protein levels of CXCL10, CXCR3, and CCR5 in orthotopic tumor tissue were increased in the low-dose combination therapy group (P <0. 05), the protein levels of CXCL10 and CXCR3 in lymph node tissue were increased in the low-dose combination therapy group (P <0. 05), the protein levels of CXCR3 and CCR5 in liver tissue were increased in the low-dose combination therapy group (P < 0. 05), and the mRNA levels of CXCR3 and CCL3 were increased in the low-dose combination therapy group (P <0.05). Compared with the low-dose combination therapy group, the mRNA and protein levels of chemokines and receptors were increased in the low-dose huangqi plus ezhu group (P < 0. 05). Conclusions The combination of Huangqi and Ezhu can inhibit the proliferation of CT26. WT orthotopic tumors, and combination therapy with 5-FU could further enhance the efficacy. The underlying therapeutic mechanism may be related to the downregulation of the CXCL10/CXCR3 and CCL3/CCR5 axes, which are closely related to tumor invasion and metastasis. [ABSTRACT FROM AUTHOR]