益气化瘀解毒方对结直肠癌循环肿瘤细胞干预作用的研究

发布时间：2018-01-13 14:39

  本文关键词：益气化瘀解毒方对结直肠癌循环肿瘤细胞干预作用的研究　出处：《湖南中医药大学》2016年博士论文　论文类型：学位论文

  更多相关文章： 益气化瘀解毒方 结直肠癌 循环肿瘤细胞 TGF-β/Snads通路

【摘要】：目的：探讨益气化瘀解毒方对HCT116-I uc2裸鼠结直肠癌模型循环肿瘤细胞的影响,揭示益气化瘀解毒方对结直肠癌循环肿瘤细胞干预作用的可能作用机制。方法：1.通过对HCT116-LUC2细胞培养及裸鼠造模,免疫磁珠筛选HCT116-luc2裸鼠结直肠癌模型循环肿瘤细胞,再用Vastern-bl ot实验方法测定成瘤后的裸鼠循环肿瘤细胞中Tβ RI、Tβ RII、smad2、smad3、smad4、smad6、 snaid7蛋白水平。2.将筛选的循环肿瘤细胞经裸鼠尾静脉注射成瘤后,观察其成瘤情况及转移率。3.随机将HCT116-luc2裸鼠结直肠癌模型分为空白模型组、益气化瘀解毒组、5-FU组,再用Vastern-bl ot实验方法测定HCT116-luc2裸鼠结直肠癌模型循环肿瘤细胞中T(3 RI xT( snaid7蛋白水平,用ELISA法检测三组成瘤裸鼠血清中TGF-(3 1含量,揭示益气化瘀解毒方对结直肠癌循环肿瘤细胞干预作用的可能作用机制,为临床提供有效的实验依据。结果：1.以EpGWligh为标记筛选HCT116-luc2裸鼠结直肠癌循环肿瘤细胞的成瘤情况良好,成瘤率达到90-%,易出现转移,出现肝转移为60%,出现肺转移为40%,肝肺同时出现转移为35%,而以EpGWlw为标记筛选的细胞不能成瘤,未出现转移,但是以gh为标记筛选的细胞仍有部分(10%)不能成瘤。2.与空白模型组比较益气化瘀解毒组能明显降低循环肿瘤细胞的Tβ RⅠ、Tβ RⅡ、Snaid4蛋白表达的水平同时升高Snad7 蛋白表达水平,两组比较具有统计学意义；与空白模型组比较5-FU组能降低Snad4 蛋白、升高Snad7 蛋白表达的水平,不能降低Tp RⅠ、Tp RTT、Snad3 蛋白表达的水平；三组比较snaid3、Snaid6比较无变化；成瘤裸鼠血清中TGF-β1含量比较：空白模型组与益气化瘀解毒组两组比较,t=5.344,P0.01,具有显著统计学意义；空白模型组与5-FU组两组比较,t=1.695,P0.05,无统计学意义。结论：1.以EpCAltfgh为标记筛选的HCT116-luc2 裸鼠结直肠癌循环肿瘤细胞成瘤情况良好易出现转移而以EpCAM为标记筛选的循环肿瘤细胞不能成瘤符合循环肿瘤细胞EpCAM为标记易成瘤的特点,但是仍有部分(10%)不能成瘤,值得进一步探讨,以EpCAM为标记筛选的循环肿瘤细胞存在假阳性率?故循环肿瘤细胞表型标准化值得进一步探讨及更深入的研究；2.益气化瘀解毒方能明显降低TβRI、Tβ RⅡ、Snad3、Snad4蛋白表达的水平同时升高Snaid7 蛋白表达水平,对snaid2、smaid6蛋白无影响,能降低成瘤裸鼠血清中TGF-β1含量,而5-FU组能降低Smad4 蛋白、升高Smad7 蛋白表达的水平,不能降低Tβ RⅠ、Tβ RⅡ、Smad3蛋白表达的水平,对smad2、smad6蛋白无影响,不能降低成瘤裸鼠血清中TGF-β1含量。益气化瘀解毒方能明显影响结直肠癌裸鼠模型循环肿瘤细胞TGF-β/Smads通路的信号强度。
[Abstract]:Objective: to investigate the effect of Yiqi Huayu jiedu recipe on circulating tumor cells in HCT116-I uc2 nude mice model of colorectal cancer. To reveal the possible mechanism of intervention of Yiqi Huayu jiedu recipe on circulating tumor cells of colorectal cancer. Methods: 1. HCT116-LUC2 cell culture and nude mice model were established. The circulating tumor cells were screened by immunomagnetic beads in HCT116-luc2 nude mice model of colorectal cancer. Vastern-bl ot assay was used to determine T 尾 RII-T 尾 -RIIad2SDA _ 3 and SMA _ 4 in circulating tumor cells of nude mice after tumorigenesis. Smad6, snaid7 protein level. 2. The selected circulating tumor cells were injected into the tail vein of nude mice to form tumor. Observe the tumorigenesis and metastasis rate of HCT116-luc2 nude mice were randomly divided into blank model group, Yiqi Huayu detoxification group and 5-FU group. Vastern-bl ot assay was used to determine the level of T3 RI xT (snaid7 protein) in circulating tumor cells of HCT116-luc2 nude mice model of colorectal cancer. ELISA method was used to detect the content of TGF-(3 1 in serum of nude mice. The possible mechanism of intervention effect of Yiqi Huayu jiedu recipe on circulating tumor cells of colorectal cancer was revealed. Results: 1. EpGWligh was used as marker to screen circulating tumor cells of colorectal cancer in HCT116-luc2 nude mice. The tumorigenesis rate reached 90-10%, easy to metastasize, liver metastasis was 60%, lung metastasis was 40%, liver and lung metastasis was 35%, and cells screened with EpGWlw as marker could not be tumorigenic. There was no metastasis, but some cells screened by gh could not be tumor-forming. 2. Compared with the blank model group, Yiqi Huayu jiedu group could significantly reduce the T 尾 R 鈪，

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