本文關(guān)鍵詞：益氣化瘀解毒方對結(jié)直腸癌循環(huán)腫瘤細胞干預(yù)作用的研究　出處：《湖南中醫(yī)藥大學(xué)》2016年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 益氣化瘀解毒方 結(jié)直腸癌 循環(huán)腫瘤細胞 TGF-β/Snads通路

【摘要】：目的：探討益氣化瘀解毒方對HCT116-I uc2裸鼠結(jié)直腸癌模型循環(huán)腫瘤細胞的影響,揭示益氣化瘀解毒方對結(jié)直腸癌循環(huán)腫瘤細胞干預(yù)作用的可能作用機制。方法：1.通過對HCT116-LUC2細胞培養(yǎng)及裸鼠造模,免疫磁珠篩選HCT116-luc2裸鼠結(jié)直腸癌模型循環(huán)腫瘤細胞,再用Vastern-bl ot實驗方法測定成瘤后的裸鼠循環(huán)腫瘤細胞中Tβ RI、Tβ RII、smad2、smad3、smad4、smad6、 snaid7蛋白水平。2.將篩選的循環(huán)腫瘤細胞經(jīng)裸鼠尾靜脈注射成瘤后,觀察其成瘤情況及轉(zhuǎn)移率。3.隨機將HCT116-luc2裸鼠結(jié)直腸癌模型分為空白模型組、益氣化瘀解毒組、5-FU組,再用Vastern-bl ot實驗方法測定HCT116-luc2裸鼠結(jié)直腸癌模型循環(huán)腫瘤細胞中T(3 RI xT( snaid7蛋白水平,用ELISA法檢測三組成瘤裸鼠血清中TGF-(3 1含量,揭示益氣化瘀解毒方對結(jié)直腸癌循環(huán)腫瘤細胞干預(yù)作用的可能作用機制,為臨床提供有效的實驗依據(jù)。結(jié)果：1.以EpGWligh為標(biāo)記篩選HCT116-luc2裸鼠結(jié)直腸癌循環(huán)腫瘤細胞的成瘤情況良好,成瘤率達到90-%,易出現(xiàn)轉(zhuǎn)移,出現(xiàn)肝轉(zhuǎn)移為60%,出現(xiàn)肺轉(zhuǎn)移為40%,肝肺同時出現(xiàn)轉(zhuǎn)移為35%,而以EpGWlw為標(biāo)記篩選的細胞不能成瘤,未出現(xiàn)轉(zhuǎn)移,但是以gh為標(biāo)記篩選的細胞仍有部分(10%)不能成瘤。2.與空白模型組比較益氣化瘀解毒組能明顯降低循環(huán)腫瘤細胞的Tβ RⅠ、Tβ RⅡ、Snaid4蛋白表達的水平同時升高Snad7 蛋白表達水平,兩組比較具有統(tǒng)計學(xué)意義；與空白模型組比較5-FU組能降低Snad4 蛋白、升高Snad7 蛋白表達的水平,不能降低Tp RⅠ、Tp RTT、Snad3 蛋白表達的水平；三組比較snaid3、Snaid6比較無變化；成瘤裸鼠血清中TGF-β1含量比較：空白模型組與益氣化瘀解毒組兩組比較,t=5.344,P0.01,具有顯著統(tǒng)計學(xué)意義；空白模型組與5-FU組兩組比較,t=1.695,P0.05,無統(tǒng)計學(xué)意義。結(jié)論：1.以EpCAltfgh為標(biāo)記篩選的HCT116-luc2 裸鼠結(jié)直腸癌循環(huán)腫瘤細胞成瘤情況良好易出現(xiàn)轉(zhuǎn)移而以EpCAM為標(biāo)記篩選的循環(huán)腫瘤細胞不能成瘤符合循環(huán)腫瘤細胞EpCAM為標(biāo)記易成瘤的特點,但是仍有部分(10%)不能成瘤,值得進一步探討,以EpCAM為標(biāo)記篩選的循環(huán)腫瘤細胞存在假陽性率?故循環(huán)腫瘤細胞表型標(biāo)準(zhǔn)化值得進一步探討及更深入的研究；2.益氣化瘀解毒方能明顯降低TβRI、Tβ RⅡ、Snad3、Snad4蛋白表達的水平同時升高Snaid7 蛋白表達水平,對snaid2、smaid6蛋白無影響,能降低成瘤裸鼠血清中TGF-β1含量,而5-FU組能降低Smad4 蛋白、升高Smad7 蛋白表達的水平,不能降低Tβ RⅠ、Tβ RⅡ、Smad3蛋白表達的水平,對smad2、smad6蛋白無影響,不能降低成瘤裸鼠血清中TGF-β1含量。益氣化瘀解毒方能明顯影響結(jié)直腸癌裸鼠模型循環(huán)腫瘤細胞TGF-β/Smads通路的信號強度。
[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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