本文選題：肝癌 + 索拉非尼�。� 參考：《浙江大學》2016年博士論文

【摘要】：背景:肝癌為我國高發(fā)惡性腫瘤之一,每年約有38.3萬人死于肝癌,約占全球肝癌死亡總數(shù)的51%。目前肝癌治療手段有限,療效不理想,病死率極高,是困擾我國群眾健康的重大問題。肝癌的常見治療手段包括外科手術(shù)、肝動脈栓塞化療術(shù)和消融術(shù)等,對于無法手術(shù)患者,化療也是重要手段之一。近年來新型分子靶向治療技術(shù)的發(fā)展為肝癌治療帶來了全新視野。靶向藥物索拉非尼可以通過抑制酪氨酸激酶阻止腫瘤新生血管的生成,還能經(jīng)Raf/MEK/ERK信號傳導通路直接抑制腫瘤生長。臨床試驗表明索拉非尼在肝癌晚期患者中展現(xiàn)了良好的療效和耐受性,但存在耐藥現(xiàn)象,其耐藥機制尚不明確,一般認為與腫瘤細胞、腫瘤基質(zhì)和個體遺傳耐藥性有關。相關研究表明肝癌耐藥相關分子的表達變化能指示患者療效敏感性,因此通過探索潛在生物標志物,建立索拉非尼治療療效的分子生物標志物譜,將有利于針對性給藥,實現(xiàn)精準醫(yī)療;此外對于關鍵分子的機制研究,將解決耐藥發(fā)生,并推動新藥研發(fā)。方法:1、外科(肝切除/肝移植)術(shù)后復發(fā)患者接受索拉非尼治療,根據(jù)影像學表現(xiàn)結(jié)合血清學指標(AFP)將入組患者分為耐藥型和敏感型。取各組肝癌及癌旁組織,同時以其他肝硬化患者及健康供肝組織作為基準對照進行iTRAQ實驗,比較耐藥組與敏感組之間組織蛋白表達差異,進而開展GO、KEGG等生物信息學分析,發(fā)現(xiàn)特征標志物譜和所涉及的生物學通路。2、建立肝癌細胞體外索拉非尼耐藥模型,在模型中采用Western blot技術(shù)對特征標志物的表達差異進一步驗證。3、利用耐藥模型,結(jié)合PPARy拮抗劑及激動劑,采用CCK8法檢測PPARγ信號通路外在干預下肝癌細胞增殖情況的變化,細胞流式技術(shù)研究細胞周期阻滯和凋亡對肝癌細胞增殖抑制的作用,探索PPARγ拮抗劑潛在的抗索拉非尼耐藥價值。結(jié)果:1、利用iTRAQ技術(shù),發(fā)現(xiàn)索拉非尼敏感組和耐藥組表達差異達到1.5倍以上的蛋白有222個,而表達差異達到2倍以上的蛋白57個,其中表達上調(diào)蛋白有26個,表達下調(diào)蛋白有31個;2、GO分析表明差異蛋白多涉及小分子代謝過程、分解代謝過程、羧酸代謝過程及含氧酸的代謝過程,而KEGG的信號通路分析同樣表明,差異蛋白富集在糖代謝、脂肪酸代謝和糖酵解通路;3、利用體外耐藥模型對2倍以上差異蛋白的驗證發(fā)現(xiàn)包括ITGA6、FN1、ICAM1、ITPA、BAX、TXNDC17和MSH2等蛋白差異與體內(nèi)結(jié)果一致,可作為機制研究的候選蛋白;4、基于生物信息學分析結(jié)果,發(fā)現(xiàn)PPARs特別是PPARγ在耐藥模型中表達發(fā)生顯著變化;5、發(fā)現(xiàn)PPARγ的拮抗劑T0070907能夠增強肝癌細胞對索拉非尼的敏感性,另一個拮抗劑GW9662未能達到同樣效果;6、,.拮抗劑T0070907及GW9662的聯(lián)用未能進一步增強細胞對索拉非尼的敏感性;7、肝癌細胞對索拉非尼敏感性的增強是細胞周期阻滯所致。結(jié)論:1、通過比較索拉非尼耐藥和敏感患者組織蛋白表達譜,發(fā)現(xiàn)多個差異蛋白,涉及脂肪酸代謝等生物學過程及信號通路;2、PPARγ可能是肝癌索拉非尼耐藥發(fā)生的關鍵因子,其拮抗劑T0070907可增強肝癌細胞的敏感性,具有潛在的臨床應用價值。
[Abstract]:Background: liver cancer is one of the high incidence of malignant tumors in China. About 383 thousand people die of liver cancer every year, accounting for about 51%. of the total number of cancer deaths in the world. At present, the treatment of liver cancer is limited, the curative effect is not ideal and the fatality rate is very high. It is a major problem perplexing the health of the masses in our country. The common treatment means of liver cancer include surgery, hepatic arterial chemoembolization and chemotherapy. In recent years, the development of new molecular targeting therapy has brought new field of vision for the treatment of liver cancer. The target drug Sola Fini can inhibit the formation of neovascularization of tumor by inhibiting tyrosine kinase, and can also suppress the swelling directly through the Raf/MEK/ERK signal transduction pathway. Tumor growth. Clinical trials show that Sola Fini has shown good efficacy and tolerance in advanced liver cancer patients, but there is a drug resistance phenomenon. The mechanism of drug resistance is not clear. It is generally considered to be related to tumor cells, tumor matrix and individual genetic resistance. Therefore, by exploring potential biomarkers and establishing molecular biomarker spectra of sorafenib treatment, it will be beneficial to targeted drug delivery and accurate medical treatment. In addition, the mechanism of key molecules will be studied to solve the drug resistance and promote new drug development. Methods: 1, surgery (hepatectomy / liver transplantation) for recurrent patients after surgery. In the treatment of sorafeni, the patients were divided into drug-resistant and sensitive types according to the imaging findings combined with serological index (AFP). The liver and paracancerous tissues were taken in each group, and the iTRAQ experiment was carried out with other liver cirrhosis patients and healthy donor liver tissue as the reference control, and the difference of tissue protein expression between the drug resistant and the sensitive groups was compared. GO, KEGG and other bioinformatics analysis, found the characteristic marker spectrum and the biological pathway involved.2, established the sorafeni drug resistance model of liver cancer cells in vitro. In the model, the expression difference of the characteristic markers was further verified by Western blot technology, and.3 was verified by the drug resistance model, PPARy antagonist and agonist, and the CCK8 method was used. The proliferation of hepatoma cells under the external intervention of PPAR gamma signaling pathway. Cell flow cytometry was used to study the effect of cell cycle arrest and apoptosis on the proliferation inhibition of hepatoma cells, and to explore the potential anti sorafeni resistance value of PPAR gamma antagonist. Results: 1, the expression difference between the current sorafenib sensitive group and the drug resistant group was 1. by using iTRAQ technique. More than 5 times more protein than 222, and the expression difference reached more than 2 times of protein 57, of which there were 26 expression up-regulated proteins and 31 down-regulated proteins; 2, GO analysis showed that the differential proteins involved small molecular metabolic processes, metabolic processes, carboxylic acid metabolism and metabolic processes of oxyacid, and KEGG signaling pathway analysis also indicated that The difference protein was enriched in sugar metabolism, fatty acid metabolism and glycolysis pathway; 3, the identification of 2 times more than the protein in vitro drug resistance model found that the differences of ITGA6, FN1, ICAM1, ITPA, BAX, TXNDC17 and MSH2 were consistent with the results of the body, and could be used as the candidate proteins for the mechanism study; 4, based on the bioinformatics analysis, the discovery of PPARs In particular, the expression of PPAR gamma was significantly changed in the drug resistance model. 5, PPAR gamma antagonist T0070907 was found to enhance the sensitivity of liver cancer cells to sorafenib, and the other antagonist GW9662 failed to achieve the same effect; 6, the combination of antagonist T0070907 and GW9662 failed to step up the cell sensitivity to sorafenib; 7, liver cancer was fine. Conclusion: 1, by comparing the tissue protein expression profiles of sorafenib and sensitive patients, a number of differential proteins are found to be involved in biological processes such as fatty acid metabolism and signaling pathways. 2, PPAR gamma may be a key factor in the occurrence of Sola Fini resistance in liver cancer, and its antagonist T007 0907, it can enhance the sensitivity of liver cancer cells and has potential clinical application value.

【學位授予單位】：浙江大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R735.7

【相似文獻】

相關期刊論文 前10條

1 丁麗;程剛;;多靶點抗腫瘤新藥索拉非尼的藥理作用及臨床研究進展[J];藥物不良反應雜志;2007年03期

2 張小麗;王鴻;;索拉非尼及其抗腫瘤作用[J];安徽醫(yī)藥;2008年03期

3 崔瑤;羅榮城;李愛民;崔斐;;索拉非尼聯(lián)合肝素抑制血管生成的研究[J];解放軍醫(yī)學雜志;2008年05期

4 逯華;陳日新;;多激酶抑制藥索拉非尼的研究進展[J];右江民族醫(yī)學院學報;2008年04期

5 孫敏;魏紅濤;蔡進;吉民;;索拉非尼的合成[J];中國藥學雜志;2009年05期

6 郭玉峰;;索拉非尼的最近研究概況[J];中國醫(yī)療前沿;2009年05期

7 陳金麟;李鐵;張沂平;;索拉非尼臨床研究進展[J];腫瘤學雜志;2009年08期

8 葉麗芬;張春紅;;口服甲苯磺酸索拉非尼患者的觀察及護理[J];臨床合理用藥雜志;2009年22期

9 劉健穎;唐偉方;陸濤;;索拉非尼研究進展[J];海峽藥學;2010年04期

10 劉韜;林子超;陳倩超;魏雪;黃偉強;黃紅兵;;索拉非尼藥物不良反應臨床特征分析與防治[J];今日藥學;2010年09期

相關會議論文 前10條

1 鄧覲云;陳悅;;索拉非尼最新研究進展[A];第十二屆全國肝癌學術(shù)會議論文匯編[C];2009年

2 劉韜;林子超;陳倩超;魏雪;黃偉強;黃紅兵;;索拉非尼藥物不良反應臨床特征分析與防治[A];2010年廣東省藥師周大會論文集[C];2011年

3 王哲;張陽;吳濤;;索拉非尼聯(lián)合伊立替康對人肝癌細胞株HepG2抑制作用的時序性依賴機制探索與研究[A];第三屆中國腫瘤內(nèi)科大會教育集暨論文集[C];2009年

4 李珍;;索拉非尼治療肝癌不良反應的觀察及護理[A];中華護理學會全國腫瘤護理新進展研討會論文匯編[C];2012年

5 莊莉;俞軍;吳健;張珉;沈恬;蔣國平;郭華;鄭樹森;;肝癌肝移植術(shù)后預防性服用索拉非尼有效改善受者生存[A];2012中國器官移植大會論文匯編[C];2012年

6 李珍;劉紅麗;張寧;;索拉非尼治療肝癌不良反應的觀察及護理[A];2012年“河南省腫瘤�？谱o士職業(yè)安全防護及新技術(shù)交流”學術(shù)會議論文集[C];2012年

7 鄭家平;邵國良;羅君;陳玉堂;姚征;曾暉;郝偉遠;;索拉非尼治療中晚期肝細胞癌安全性和生存因素分析[A];2013年浙江省放射學學術(shù)年會論文集[C];2013年

8 劉淼;;索拉非尼常見不良反應及對策[A];中國成人醫(yī)藥教育論壇（2009）[C];2009年

9 趙振宇;戈偉;;索拉非尼靶向治療非小細胞肺癌研究進展[A];第二屆湖北省腫瘤靶向治療學術(shù)會議論文選[C];2007年

10 成炳祥;方煊;朱承良;婁s，

本文編號：1817819