黄芪—莪术配伍联合奥沙利铂对CT26.WT原位移植瘤小鼠中趋化因子及其受体表达的影响

发布时间：2018-12-15 17:28

【摘要】：[目的]本研究采用手术手段建立Balb/c小鼠结肠癌原位移植瘤模型,探讨中药扶正祛邪药对——黄芪-莪术水提物与奥沙利铂(L-OHP)联合应用抑制鼠结肠癌CT26 wild-type(CT26.WT)细胞的转移,并调控CXCR3、CCR6、CXCL9、CCL20在mRNA及蛋白水平的表达。[方法]1.建立Balb/c小鼠结肠癌原位移植瘤模型,观察各组肿瘤在小鼠体内的生长情况,初步观察小鼠肝脏转移的情况。2.通过黄芪-莪术分别以高、中、低剂量联合L-OHP对小鼠治疗,观察用药后小鼠体重,瘤体大小,肝脏转移情况的变化。3.采用Real time-PCR方法分别检测原位移植肿瘤灶与肝转移灶CXCR3、CCR6、CXCL9、CCL20mRNA 的表达情况。4.采用Western Blotting方法分别检测原位移植肿瘤灶与肝转移灶CXCR3、CCR6、CXCL9、CCL20的蛋白表达情况。[结果]1.成功建立Balb/c小鼠结肠癌原位移植瘤模型,成模率达到100%。2.L-OHP单独应用和联合中药应用均能抑制原位移植瘤体的增大,阳性药抑制原位移植瘤生长的效果最好(P0.01)。联合用药能降低肝转移灶的发生(P0.05)。3.阳性药组及联合用药低剂量组CXCR3及CCR6mRNA的表达显著下调(P0.01)。用药组CXCL9、CCL20mRNA表达未见统计学意义。4.与空白对照组相比,联合用药能不同程度下调CXCR3、CCR6蛋白表达(P0.05,P0.01)。在肝组织内,联合用药可以下调CXCL9、CCL20蛋白表达(P0.01)。[结论]L-OHP联合黄芪-莪术作用于荷瘤小鼠能够显著地抑制原位移植结肠癌的肝转移,降低趋化因子CXCR3、CCR6蛋白与mRNA的表达,低剂量联合效果最显著,具有浓度依赖性。中药联合化疗药抗肿瘤转移机制可能与下调CXCR3、CCR6蛋白与mRNA相关,对CXCL9、CCL20蛋白与mRNA的影响尚待进一步实验证明。CXCR3、CCR6可成为判断结肠癌预后的潜在靶点。
[Abstract]:[objective] to establish an orthotopic transplanted tumor model of colon cancer in Balb/c mice by surgical method. Objective: to investigate the effect of traditional Chinese medicine (TCM) on inhibiting the metastasis of rat colon cancer CT26 wild-type (CT26.WT) cells and regulating the expression of CXCR3,CCR6,CXCL9,CCL20 in mRNA and protein levels by combined application of Astragalus curcumae aqueous extract and oxaliplatin (L-OHP). [methods] 1. The orthotopic transplanted tumor model of colon cancer in Balb/c mice was established to observe the growth of tumor in mice and the liver metastasis in mice. 2. The mice were treated with high, middle and low doses of Astragalus curcumae combined with L-OHP respectively. The changes of body weight, tumor size and liver metastasis were observed after administration of Astragalus curcumae. 3. Real time-PCR method was used to detect the expression of CXCR3,CCR6,CXCL9,CCL20mRNA in orthotopic transplantation tumor and liver metastasis respectively. 4. Western Blotting method was used to detect the expression of CXCR3,CCR6,CXCL9,CCL20 protein in tumor focus and liver metastases of orthotopic transplantation. [result] 1. The orthotopic transplanted tumor model of colon cancer in Balb/c mice was successfully established. The rate of model formation reached that of 100%.2.L-OHP alone and combined with traditional Chinese medicine (TCM), both of which could inhibit the enlargement of orthotopic transplantation tumor. The positive drug had the best effect on tumor growth in orthotopic transplantation (P0.01). Combined use of drugs can reduce the incidence of liver metastasis (P0.05). The expression of CXCR3 and CCR6mRNA in positive group and low dose group was significantly decreased (P0.01). The expression of CXCL9,CCL20mRNA in the medication group was not statistically significant. 4. 4. Compared with the control group, the expression of CXCR3,CCR6 protein was down-regulated (P0.05, P0.01). In liver tissue, the expression of CXCL9,CCL20 protein could be down-regulated by combined medication (P0.01). [conclusion] L-OHP combined with astragalus curcuma can significantly inhibit liver metastasis and decrease the expression of chemokine CXCR3,CCR6 protein and mRNA in tumor-bearing mice. The mechanism of anti-tumor metastasis of traditional Chinese medicine combined with chemotherapy may be related to down-regulation of CXCR3,CCR6 protein and mRNA, and the effect on CXCL9,CCL20 protein and mRNA remains to be proved by further experiments. CXCR3,CCR6 may be a potential target for judging the prognosis of colon cancer.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R73-36

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