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  本文選題：艾迪注射液 + 多藥耐藥　； 參考：《貴州醫(yī)科大學》2017年碩士論文

【摘要】：目的:探討艾迪注射液對人肝癌多藥耐藥細胞Bel-7402/5-FU的耐藥性是否具有逆轉作用及其可能的逆轉機制。方法:1.采用5-氟尿嘧啶(fluorouracil,5-FU)通過濃度梯度遞加法建立人肝癌耐藥細胞Bel-7402/5-FU。2.待耐藥細胞Bel-7402/5-FU建立成功后,將實驗分為兩組,即Bel-7402組和Bel-7402/5-FU組,將兩組細胞種植于96孔板中,待細胞貼壁穩(wěn)定后,再將每種細胞分為7個小組,每種細胞的第一組加入正常培養(yǎng)基,后面六組分別加入含5-FU、ADM、MMC、MTX、CTX、CDDP的培養(yǎng)基,每孔終體積為200ul,培養(yǎng)24h后,每孔加入10ul的cck-8試劑,繼續(xù)培養(yǎng)2h后,酶標儀在450nm下讀取吸光度值,使用SPSS19.0統(tǒng)計軟件計算每種化療藥物對親本細胞Bel-7402及耐藥細胞Bel-7402/5-FU 的半數(shù)抑制濃度(half maximal inhibitory concentration,IC50),計算6種化療藥物Bel-7402細胞和Bel-7402/5-FU細胞的抑制率(IR)和Bel-7402/5-FU的耐藥指數(shù)(RI)。3.通過不同濃度的艾迪注射液作用于Bel-7402/5-FU后,cck-8法檢測艾迪注射液對Bel-7402/5-FU是否具有抑制作用。4.將實驗分為3組,即Bel-7402組、Bel-7402/5-FU組和艾迪注射液作用過的Bel-7402/5-FU組,分別提取三組細胞中的總蛋白后,Western blot法檢測三組細胞中多藥耐藥相干蛋白1(MRP1)、P-糖蛋白(P-gp,170kD)、程序化死亡因子5(PDCD5,15kD)蛋白的表達情況。結果:1、Bel-7402/5-FU細胞對6種化療藥物的半數(shù)抑制濃度較Bel-7402細胞明顯升高,對6種化療藥物表現(xiàn)出不同程度的耐藥性。2、艾迪注射液對Bel-7402/5-FU細胞具有抑制作用,且隨著艾迪注射液濃度的增加,抑制作用越強。3、與Bel-7402細胞相比,Bel-7402/5-FU細胞株中MRP1、P-gp表達明顯升高,PDCD5表達較低。使用半數(shù)抑制濃度的艾迪注射液分別作用于Bel-7402細胞及Bel-7402/5-FU細胞后,與Bel-7402/5-FU細胞相比,經(jīng)過艾迪注射液作用后的Bel-7402/5-FU細胞中MRP1、P-gp表達降低,PDCD5表達增高,差異具有統(tǒng)計學意義(P0.05)。結論:艾迪注射液對肝癌細胞的多藥耐藥性具有一定的逆轉作用,其機制與降低多藥耐藥相關蛋白1和P-糖蛋白的表達,促進凋亡相關蛋白5的表達有關。
[Abstract]:Aim: to investigate whether Aidi injection can reverse the drug resistance of multidrug resistant human hepatoma cell line Bel-7402/5-FU and its possible reversal mechanism. Method 1: 1. 5-fluorouraciline 5-FU (5-FU) was used to establish human hepatoma resistant cell line Bel-7402 / 5-FU. After the establishment of drug-resistant cell line Bel-7402/5-FU, the experiment was divided into two groups: Bel-7402 group and Bel-7402/5-FU group. The two groups of cells were implanted in 96-well plate. When the cells were stably adhered to the wall, each cell was divided into seven groups. The first group of each cell was added to the normal medium, and the latter six groups were added respectively to the medium containing 5-FUU ADMU MMCMTXTX CTX CDDP. The final volume of each pore was 200 ul.After 24 hours of culture, the cck-8 reagent of 10ul was added to each pore, and after 2 hours of culture, the absorbance value was read by the enzyme marker under 450nm. SPSS19.0 software was used to calculate the half maximal inhibitory concentration (IC50) of each chemotherapeutic drug on Bel-7402 and Bel-7402/5-FU, and to calculate the inhibitory rate of Bel-7402 and Bel-7402/5-FU cells on 6 chemotherapeutic drugs (Bel-7402 and Bel-7402/5-FU) and the resistance index of Bel-7402/5-FU (RIN. 3). The inhibitory effect of Aidi injection on Bel-7402/5-FU was detected by cck-8 method after different concentrations of Aidi injection was treated with Bel-7402/5-FU. The experiment was divided into three groups: Bel-7402 group, Bel-7402 / 5-FU group and Bel-7402/5-FU group, which were treated with Aidi injection. The total protein was extracted from the three groups of cells by Western blot method to detect the expression of multidrug resistance-associated protein (MRP1P-1), P-gpfU (170kDN) and programmed death factor 5PDCD5( 15kD) in the three groups of cells. Results the half inhibitory concentration of Bel-7402 / 5-FU cells to 6 chemotherapeutic drugs was significantly higher than that of Bel-7402 cells, and the drug resistance to 6 chemotherapeutic drugs was different. Adi injection had inhibitory effect on Bel-7402/5-FU cells, and with the increase of Aidi injection concentration, The stronger the inhibitory effect was, the lower the expression of PDCD5 was in Bel-7402 / 5-FU cell line compared with that in Bel-7402 cell line. Compared with Bel-7402/5-FU cells, the expression of MRP1P-gp in Bel-7402/5-FU cells treated with Aidi injection with 50% inhibitory concentration was significantly lower than that in Bel-7402/5-FU cells, and the difference was statistically significant (P 0.05). Conclusion: Aidi injection can reverse the multidrug resistance of hepatoma cells, and its mechanism is related to the reduction of the expression of multidrug resistance-associated protein 1 and P-glycoprotein, and the promotion of apoptosis-related protein 5 expression.
【學位授予單位】：貴州醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.7

【參考文獻】

相關期刊論文 前10條

1 楊柳青;秦叔逵;趙寧莉;華海清;劉秀峰;陳映霞;王琳;朱艷;;FOLFOX4方案治療中晚期原發(fā)性肝癌的臨床研究[J];臨床腫瘤學雜志;2013年02期

2 田思源;徐愛兵;沈茜;蔡鴻宇;周元;;吉西他濱聯(lián)合奧沙利鉑治療晚期原發(fā)性肝癌的臨床觀察[J];腫瘤基礎與臨床;2012年01期

3 楊盛力;張小玲;劉利平;熊枝繁;曹仕瓊;郭鋒偉;;乙肝病毒X蛋白通過激活NF-κB信號通路上調肺耐藥相關蛋白的表達[J];華中科技大學學報(醫(yī)學版);2012年01期

4 田彥璋;趙浩亮;賀杰峰;李輝宇;劉宏;韓建立;;葡萄糖神經(jīng)酰胺合成酶在肝癌細胞多藥耐藥中的作用機制[J];中華實驗外科雜志;2011年11期

5 ;中藥逆轉肝癌多藥耐藥的研究現(xiàn)狀(英文)[J];Chinese-German Journal of Clinical Oncology;2011年09期

6 錢軍;秦叔逵;;原發(fā)性肝癌的系統(tǒng)性化療[J];消化腫瘤雜志(電子版);2009年02期

7 王映梅;黃高f;惠延平;張靜;;乳腺癌耐藥相關蛋白在肝硬化及肝癌組織中的表達[J];中華肝臟病雜志;2009年03期

8 楊盛力;彭靜;陳孝平;;中藥逆轉肝癌多藥耐藥的研究進展[J];肝膽外科雜志;2008年06期

9 左小東;鄔曉敏;崔永安;秦叔逵;金成;;吉西他濱聯(lián)合奧沙利鉑治療原發(fā)性肝癌5例報告[J];臨床腫瘤學雜志;2008年12期

10 張雋開;王忠裕;丁大朋;辛毅;;黃芪對肝癌耐藥細胞株Bel/Fu化療敏感性的影響[J];中國中西醫(yī)結合外科雜志;2008年04期

相關博士學位論文 前1條

1 羅順峰;低糖微環(huán)境對肝癌細胞多藥耐藥的影響[D];華中科技大學;2006年

，

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