黄芪莪术联合5-氟尿嘧啶对CT26.WT原位移植瘤小鼠CXCL10/CXCR3轴和CCL3/CCR5轴表达的影响

目的 探讨黄芪莪术配伍联合5-氟尿嘧啶(5-FU)治疗结肠癌的作用机制.方法 采用1只BALB/c雄性小鼠作为种鼠,生成原位瘤体,80只BALB/c雄性小鼠采用移植种鼠瘤体的方法建立CT26.WT原位移植瘤小鼠模型.将80只BALB/c雄性小鼠随机分为模型组、5-FU组(25 mg/kg)、黄芪莪术高剂量组(12 g/...

Combined effects of Huangqi and Ezhu with 5-fluorouracil on CXCL10/CXCR3 and CCL3/CCR5 expression in mice with CT26.WT orthotopic transplanted tumor

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(3 g/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 and 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.