本文選題：肝細(xì)胞癌 + 患者來(lái)源的移植瘤模型�。� 參考：《第二軍醫(yī)大學(xué)》2016年博士論文

【摘要】：背景:雖然近幾年外科及其他治療手段的不斷進(jìn)步,但是肝癌患者總體生存率依舊不理想,原因在于肝癌患者發(fā)病隱匿,早期診斷困難,多數(shù)患者在確診時(shí)已為晚期,失去手術(shù)機(jī)會(huì);即使行手術(shù)治療,術(shù)后仍有較高的復(fù)發(fā)率,且復(fù)發(fā)后缺乏有效治療手段。因此,尋找對(duì)晚期肝癌和復(fù)發(fā)性肝癌切實(shí)有效的治療方法,對(duì)延長(zhǎng)肝癌患者生存有重要意義。分子靶向藥在腫瘤治療中的應(yīng)用越來(lái)越多,索拉菲尼也被證實(shí)在肝癌中有效,提示分子靶向藥物可能是治療肝癌的新方法。但是目前進(jìn)入肝癌臨床試驗(yàn)的靶向藥物中,除了索拉菲尼以外,其余全部被證實(shí)在總體人群中無(wú)效。即使是索拉菲尼,在臨床應(yīng)用中同樣有約50%的患者無(wú)效,但是我們無(wú)法在用藥之前判斷索拉菲尼的療效。因此,肝癌缺乏有效的分子靶向藥物和用來(lái)指導(dǎo)靶向藥物治療的生物標(biāo)志物或者分子分型�；颊邅�(lái)源的移植瘤模型(Patient-derived xenograft models,PDXs)是目前被認(rèn)為最接近臨床患者的藥物篩選模型。PDX模型保持了所來(lái)源腫瘤的生物學(xué)特征,并且PDX對(duì)藥物的療效評(píng)價(jià)與藥物臨床實(shí)際效果較為一致,逐漸地被應(yīng)用于藥物研發(fā)、敏感標(biāo)志物篩選等研究中。但是肝癌PDX模型和其對(duì)臨床患者腫瘤的代表性報(bào)道相對(duì)較少,其應(yīng)用價(jià)值值得探討。索拉菲尼作為唯一應(yīng)用于肝癌臨床治療的藥物,在整體人群中僅有約50%患者有效,而另一半患者則對(duì)索拉菲尼不敏感,使得索拉菲尼的整體療效欠佳,并使得一半患者承受藥物不良反應(yīng)帶來(lái)的痛苦和巨大的經(jīng)濟(jì)負(fù)擔(dān)。而在索拉菲尼耐藥研究中,受限于體內(nèi)外腫瘤模型的限制,如體外模型多,體內(nèi)模型少,模式動(dòng)物多,人源化的模型少,使得索拉菲尼耐藥機(jī)制和標(biāo)志物仍不清楚,亟待進(jìn)一步研究。方法:本研究首先通過(guò)新鮮腫瘤組織移植方法構(gòu)建了大規(guī)模的肝癌PDX模型隊(duì)列,并對(duì)比PDX模型與對(duì)應(yīng)來(lái)源腫瘤組織的生物學(xué)特征,通過(guò)全外顯子組測(cè)序、SNP芯片、表達(dá)譜芯片等技術(shù),描繪了PDX模型的分子遺傳學(xué)特征,并與現(xiàn)有肝癌測(cè)序數(shù)據(jù)對(duì)比,來(lái)研究PDX模型與來(lái)源患者腫瘤組織的生物學(xué)一致性。同時(shí)對(duì)建模成功與未建成功患者的臨床資料進(jìn)行對(duì)比分析,分析建模過(guò)程本身的應(yīng)用價(jià)值。并且根據(jù)PDX模型遺傳學(xué)特征進(jìn)行可能用于肝癌治療的靶向藥物整理和初步篩選。通過(guò)聯(lián)合模型對(duì)索拉菲尼的敏感性與表達(dá)譜數(shù)據(jù),尋找與索拉菲尼藥物敏感性相關(guān)的生物標(biāo)志物,并通過(guò)體內(nèi)外實(shí)驗(yàn)驗(yàn)證。結(jié)果:本研究共建成肝細(xì)胞癌PDX模型242例、肝內(nèi)膽管癌PDX模型71例,建模成功率分別為30.2%和33.8%,平均建成時(shí)間為90(65,130)天和110(80,150)天。PDX建模成功所需的時(shí)間短于相應(yīng)患者的無(wú)瘤生存期。PDX模型保持了所來(lái)源腫瘤組織的病理學(xué)特點(diǎn),并且在腫瘤標(biāo)志物甲胎蛋白(alpha fetoprotein,AFP)的表達(dá)上與患者一致。建模成功在肝細(xì)胞癌患者中與患者年齡50、HBe Ag陽(yáng)性、腫瘤直徑5cm、AFP20U/L、Edmondson GradeⅢⅣ級(jí)、微血管侵犯、大血管侵犯、子灶、包膜不完整、早期復(fù)發(fā)等惡性表型相關(guān),并且預(yù)示患者較差的無(wú)瘤生存期及總體生存期。同樣建模成功在肝內(nèi)膽管癌中與CA19-939U/L、早期復(fù)發(fā)等指標(biāo)相關(guān),也預(yù)示肝內(nèi)膽管癌較差的臨床預(yù)后。肝癌患者常見(jiàn)的基因突變中,除少數(shù)基因的變異未在PDX模型中檢測(cè)出,絕大多數(shù)均可在PDX模型中找到。在PDX模型中肝癌常見(jiàn)突變基因的突變率分別為T(mén)P53(67.62%)、TP53(67.6%)、TTN(93.3%)、CTNNB1(11.4%)、JAK1(6.7%)等;常見(jiàn)拷貝數(shù)擴(kuò)增ARNT(20.5%)、MTDH(16.2%)、CCND1(12.4%)、FGF19(11.4%)和MET(16.1%)等;拷貝數(shù)缺失RB1(11.4%)、SMAD4(18.1%)、WRN(15.2%)、ARID1A(7.6%)、CDKN2A(12.4%)和PTEN(5.7%)等。索拉菲尼在PDX模型中的有效率約為40%,與臨床試驗(yàn)的結(jié)果相符。對(duì)索拉菲尼耐藥的PDX模型中細(xì)胞周期、WNT信號(hào)通路、IFNa/b信號(hào)通路異常,聯(lián)合應(yīng)用細(xì)胞周期的抑制劑LEE011及IFNa均可提高索拉菲尼抗腫瘤效果。DKK1在耐藥組中高表達(dá),并且在體外誘導(dǎo)耐藥細(xì)胞系中高表達(dá),同時(shí)在PDX模型中與腫瘤較差的無(wú)進(jìn)展生存期相關(guān),且在連續(xù)用藥過(guò)程中上調(diào)。干擾DKK1后可影響細(xì)胞對(duì)索拉菲尼的敏感性。臨床上,用藥一年內(nèi)死亡的患者血清DKK1水平高于用藥一年以上的患者。結(jié)論:1.肝癌PDX模型保持了患者腫瘤組織的生物學(xué)特點(diǎn),肝癌常見(jiàn)的基因突變均可在PDX模型隊(duì)列中找到對(duì)應(yīng)模型,PDX模型建成時(shí)間早于腫瘤的復(fù)發(fā)時(shí)間,PDX建成功意味著患者較差的臨床預(yù)后,需進(jìn)行密切隨訪及積極干預(yù)。2.索拉菲尼耐藥過(guò)程中細(xì)胞周期、WNT信號(hào)通過(guò)發(fā)揮重要作用,聯(lián)合應(yīng)用這些信號(hào)通路抑制劑可增強(qiáng)索拉菲尼療效。DKK1是索拉菲尼耐藥的潛在標(biāo)志物,并且干擾DKK1可增強(qiáng)腫瘤對(duì)索拉菲尼的敏感性。
[Abstract]:Background: Despite the continuous progress of surgery and other treatments in recent years, the overall survival rate of the patients with liver cancer is still not ideal. The reason lies in the concealment of the patients with liver cancer and the difficulty of early diagnosis. Most patients are late at the time of diagnosis and lose the chance of operation. Even if the operation is performed, there is still a high recurrence rate after the operation, and the recurrence is lack after the recurrence. Therefore, it is of great significance to search for the effective treatment of advanced liver cancer and recurrent liver cancer, which is of great significance for prolonging the survival of the patients with liver cancer. More and more applications of molecular targeting drugs are used in the treatment of cancer. Sola Feeney has also been proved to be effective in liver cancer, suggesting that molecular targeting drugs may be a new method for the treatment of liver cancer. At present, all the target drugs that enter the clinical trial of liver cancer, except Sola Feeney, are all proved to be ineffective in the general population. Even Sola Feeney, about 50% of the patients are ineffective in clinical application, but we can not judge the effect of Sola Feeney before using the drug. Therefore, the liver cancer lacks effective molecular targeting drugs. And the biomarkers or molecular types used to guide targeted drug therapy. The Patient-derived xenograft models (PDXs) is the drug screening model that is currently considered the closest to the clinical patients. The.PDX model of the drug screening model maintains the biological characteristics of the cancer, and the evaluation of the efficacy of PDX on the drug and the presence of the drug. The actual effect of the bed is more consistent and is gradually used in the research of drug development and sensitive marker screening. However, the PDX model of liver cancer and its representative reports on the clinical patients are relatively less, and its application value is worth discussing. As the only drug that should be used for the clinical treatment of liver cancer, only about 50% of the whole population are affected by Sola Feeney. The other half of the patients were insensitive to Sola Feeney, which made the overall efficacy of Sola Feeney poor and made half of the patients suffering from the pain and enormous economic burden of adverse drug reactions. In Sola Feeney's drug resistance study, the restriction on the tumor model in the body and outside the body, such as in vitro models, less models in the body, and pattern movement, was limited. The mechanisms and markers of Sola Feeney resistance are still unclear. It is not clear that the drug resistance mechanism and markers are still unclear. Methods: first of all, a large PDX model of liver cancer was constructed by the method of fresh tumor tissue transplantation, and the biological characteristics of the PDX model and the corresponding source of the tumor tissue were compared, and the whole exons were sequenced. SNP chip, expression spectrum chip and other techniques, describe the molecular genetic characteristics of the PDX model, and compare with the existing liver cancer sequencing data to study the biological consistency of the PDX model and the cancer tissue from the source of the patients. The clinical data of the successful modeling and the unsuccessful patients are compared and analyzed, and the application value of the modeling process itself is analyzed. And according to the PDX model genetic characteristics to carry out the targeted drug arrangement and preliminary screening for the treatment of liver cancer. Through the combined model of Sola Feeney's sensitivity and expression data, look for biomarkers related to the sensitivity of Sola Feeney drug, and verify it through the test in vivo and in vitro. Results: this study was built into hepatocellular carcinoma PDX In 242 cases and 71 cases of intrahepatic cholangiocarcinoma PDX model, the successful rate of modeling was 30.2% and 33.8%, the average construction time was 90 (65130) days and 110 (80150) days. The time needed for the success of.PDX modeling was shorter than that of the corresponding patient's tumor free survival.PDX model, which kept the pathological features of the tumor tissue, and the tumor marker alpha fetoprotein (alph). The expression of a fetoprotein, AFP) was consistent with the patient. The modeling was successful in patients with hepatocellular carcinoma with age 50, HBe Ag positive, tumor diameter 5cm, AFP20U/L, Edmondson Grade III IV, microvascular invasion, large vascular invasion, subfoci, incomplete capsule, early recurrence and other malignant phenotype related, and predict the patient's poor tumor free survival and the prognosis. The same modeling success is associated with CA19-939U/L and early recurrence in intrahepatic cholangiocarcinoma. It also indicates the poor clinical prognosis of intrahepatic cholangiocarcinoma. Most of the common gene mutations in the liver cancer patients are found in the PDX model except that a few genes are not detected in the PDX model. In the PDX model, the liver cancer is common. The mutation rates of the mutant genes were TP53 (67.62%), TP53 (67.6%), TTN (93.3%), CTNNB1 (11.4%), JAK1 (6.7%), common copy number amplification ARNT (20.5%), MTDH (16.2%), CCND1 (12.4%), FGF19 (11.4%) and MET (16.1%), SMAD4 (18.1%), 15.2%, 15.2%, 15.2%, 16.1%, etc. The effective rate in the model is about 40%, which is consistent with the results of clinical trials. The cell cycle, WNT signaling pathway, IFNa/b signaling pathway in the Sola Feeney resistant PDX model, the combination of LEE011 and IFNa, the combined use of cell cycle inhibitors, LEE011 and IFNa, can increase the high expression of sorafeni's anti-tumor effect in the drug resistant group, and induce drug resistance in vitro. High expression in the cell line was associated with a poor progression free survival in the PDX model and increased during continuous use. Interfering with DKK1 could affect the sensitivity of the cell to Sola Feeney. In clinical, the serum DKK1 level of patients who died within one year was higher than that of patients who had been used for more than one year. Conclusion: 1. the PDX model of liver cancer has been maintained. The biological characteristics of the tumor tissues of the patients, the common gene mutation of the liver cancer can be found in the PDX model queue. The time of the PDX model is earlier than the time of the tumor recurrence. The success of PDX means that the patient has poor clinical prognosis. It is necessary to follow up closely and actively intervene in the cell cycle during the process of.2. Sola Feeney resistance, and the WNT signal is connected. The combined use of these signaling pathways can enhance the efficacy of the Sola Feeney effect.DKK1 as a potential marker for Sola Feeney resistance, and interfering with DKK1 can enhance the sensitivity of the tumor to Sola Feeney.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R735.7

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王勇姿,俞超芹,凌昌全,衛(wèi)立辛;肝癌細(xì)胞系的研究概況[J];上海醫(yī)學(xué);2000年02期

2 王蓬,陳孝平,PBSLai,CK Leow,Joseph W Y Lau;二十碳五烯酸對(duì)三種肝癌細(xì)胞系作用的實(shí)驗(yàn)研究[J];中華普通外科雜志;2001年01期

3 彭林,王衛(wèi)東,孫建,簡(jiǎn)志祥,杜嘉林,區(qū)金銳;穩(wěn)定表達(dá)促血管生成素基因肝癌細(xì)胞系的建立及產(chǎn)物的檢測(cè)[J];廣東醫(yī)學(xué);2003年07期

4 趙蕩;陳宏武;許磊;劉飛勇;;不同時(shí)期肝癌免疫殺傷腫瘤細(xì)胞效應(yīng)探討[J];中山大學(xué)學(xué)報(bào)(醫(yī)學(xué)科學(xué)版);2006年S1期

5 郭曉林;姜雅秋;金清龍;;多藥耐藥相關(guān)蛋白-3表達(dá)與肝癌耐藥的相關(guān)性[J];中國(guó)老年學(xué)雜志;2010年11期

6 ;離體培養(yǎng)的人體肝癌細(xì)胞系的建立[J];醫(yī)學(xué)研究通訊;1978年10期

7 陸榮華;陳尊器;陳瑞銘;;人體肝癌細(xì)胞系體外惡性浸潤(rùn)行為的研究[J];細(xì)胞生物學(xué)雜志;1982年02期

8 張志培,施新猷,李六金,劉雪松,江遜,彭磊,趙佐慶;小鼠免疫力對(duì)接種人類肝癌細(xì)胞系9724生長(zhǎng)的影響[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2001年07期

9 張志仁,徐順清,周宜開(kāi),任恕,劉志偉,呂斌,徐永俊;一株受二VA英類化學(xué)物質(zhì)誘導(dǎo)表達(dá)的螢光素酶肝癌細(xì)胞系[J];中國(guó)生物化學(xué)與分子生物學(xué)報(bào);2001年06期

10 董琳,王桂珍;從分子基因水平開(kāi)展中醫(yī)藥防治肝癌的研究[J];甘肅中醫(yī);2002年04期

相關(guān)會(huì)議論文 前10條

1 符兆英;劉凱歌;徐光華;;肝癌細(xì)胞系中乙型肝炎病毒x蛋白與缺氧誘導(dǎo)因子-1α表達(dá)的研究[A];中國(guó)細(xì)胞生物學(xué)學(xué)會(huì)2005年學(xué)術(shù)大會(huì)、青年學(xué)術(shù)研討會(huì)論文摘要集[C];2005年

2 張朵;朱海英;謝東莆;胡以平;;肝癌細(xì)胞中存在SP細(xì)胞[A];中國(guó)細(xì)胞生物學(xué)學(xué)會(huì)2005年學(xué)術(shù)大會(huì)、青年學(xué)術(shù)研討會(huì)論文摘要集[C];2005年

3 汪謙;曹良啟;陳錫林;甄茂川;黃曉卉;傅新暉;;過(guò)氧化物酶增殖物激活受體活化抑制肝癌細(xì)胞遷移的研究[A];廣東省肝臟病學(xué)會(huì)2007年年會(huì)論文集[C];2007年

4 鄧欣;蔣小玲;李輝;吳其愷;聶廣;劉欽;;表達(dá)耐拉米夫定乙型肝炎病毒肝癌細(xì)胞系的建立[A];第一次全國(guó)中西醫(yī)結(jié)合傳染病學(xué)術(shù)會(huì)議論文匯編[C];2006年

5 吳塵軒;杜智;朱爭(zhēng)艷;高英堂;王毅軍;只丹琮;孫泉;;肝癌免疫治療微環(huán)境和樹(shù)突狀細(xì)胞體外誘導(dǎo)的實(shí)驗(yàn)研究[A];天津市生物醫(yī)學(xué)工程學(xué)會(huì)2007年學(xué)術(shù)年會(huì)論文摘要集[C];2007年

6 趙主江;單云峰;樊紅;程欲超;成建;權(quán)艷梅;張建瓊;謝維;;DNMT3A siRNA穩(wěn)定表達(dá)載體的構(gòu)建及其沉默效率的鑒定[A];江蘇省遺傳學(xué)會(huì)第七屆代表大會(huì)暨學(xué)術(shù)研討會(huì)論文摘要匯編[C];2006年

7 田德安;;RhoA短發(fā)夾狀雙鏈RNA對(duì)肝癌細(xì)胞生物學(xué)行為的調(diào)控研究[A];中華醫(yī)學(xué)會(huì)第七次全國(guó)消化病學(xué)術(shù)會(huì)議論文匯編（下冊(cè)）[C];2007年

8 高倩;;肝癌HepG2細(xì)胞系分泌的免疫抑制物質(zhì)的研究[A];第六屆全國(guó)免疫學(xué)學(xué)術(shù)大會(huì)論文集[C];2008年

9 代智;劉銀坤;馮巨濤;宋海燕;申華麗;張麗君;崔杰峰;;高低轉(zhuǎn)移潛能肝癌細(xì)胞系差異糖蛋白質(zhì)組學(xué)研究[A];中國(guó)蛋白質(zhì)組學(xué)第二屆學(xué)術(shù)大會(huì)論文摘要論文集[C];2004年

10 張盛周;俞明月;謝晶;戴谷;李朝軍;;Met與Fas基因在肝癌組織和肝癌細(xì)胞系中的表達(dá)[A];中國(guó)細(xì)胞生物學(xué)學(xué)會(huì)第八屆會(huì)員代表大會(huì)暨學(xué)術(shù)大會(huì)論文摘要集[C];2003年

相關(guān)重要報(bào)紙文章 前1條

1 記者 胡德榮;肝癌患者自噬性缺陷表型與預(yù)后不良緊密相關(guān)[N];健康報(bào);2008年

相關(guān)博士學(xué)位論文 前10條

1 王驥;凝集素富集糖肽聯(lián)合酶促~(18)O_3標(biāo)記的定量N-糖蛋白組學(xué)方法在肝癌術(shù)后轉(zhuǎn)移復(fù)發(fā)研究中的應(yīng)用[D];復(fù)旦大學(xué);2012年

2 劉坤;Bcl-2/Twist1蛋白互作在缺氧促進(jìn)肝癌血管生成擬態(tài)形成中的作用[D];天津醫(yī)科大學(xué);2015年

3 陳勇;蓽茇酰胺抑制肝癌細(xì)胞遷移/侵襲的作用及其機(jī)制[D];華中科技大學(xué);2015年

4 林鎮(zhèn)海;NR1D1通過(guò)DUOX2/MCP-1途徑募集巨噬細(xì)胞促進(jìn)肝癌侵襲轉(zhuǎn)移的機(jī)制及臨床研究[D];復(fù)旦大學(xué);2014年

5 丁坤;骨橋蛋白促進(jìn)肝癌細(xì)胞順鉑耐藥的實(shí)驗(yàn)研究[D];山東大學(xué);2016年

6 馬春亮;KLF17在肝癌中的作用及對(duì)肝癌預(yù)后的影響[D];北京協(xié)和醫(yī)學(xué)院;2016年

7 幸思忠;URG4/URGCP激活NF-κB通路調(diào)控肝細(xì)胞癌血管增生的實(shí)驗(yàn)研究[D];南方醫(yī)科大學(xué);2016年

8 范婭涵;Hedgehog信號(hào)通路調(diào)控Twist1表達(dá)促進(jìn)肝癌細(xì)胞侵襲轉(zhuǎn)移分子機(jī)制研究[D];第三軍醫(yī)大學(xué);2016年

9 張素潔;長(zhǎng)非編碼RNA LncTSG1抑制肝癌轉(zhuǎn)移的功能及機(jī)制研究[D];中國(guó)人民解放軍醫(yī)學(xué)院;2016年

10 梁東;Fbxw7促進(jìn)Slug降解抑制TGF-β誘導(dǎo)的肝癌轉(zhuǎn)移[D];福建醫(yī)科大學(xué);2015年

相關(guān)碩士學(xué)位論文 前10條

1 田蕓蕓;人臍帶MSCs上清對(duì)肝癌細(xì)胞系HepG2影響的比較轉(zhuǎn)錄組學(xué)研究[D];內(nèi)蒙古大學(xué);2015年

2 徐辰;NKCC1促進(jìn)肝癌細(xì)胞轉(zhuǎn)移的分子機(jī)制及其抑制劑用于抗腫瘤藥物的探索[D];安徽醫(yī)科大學(xué);2015年

3 楊霞;循環(huán)肝癌細(xì)胞CD147表型鑒定在指導(dǎo)利卡汀個(gè)體化治療中的應(yīng)用潛能探索[D];蘇州大學(xué);2015年

4 石志勇;FOXF2在肝癌中的表達(dá)及其對(duì)預(yù)后價(jià)值的研究[D];蘇州大學(xué);2015年

5 陳楊;缺氧誘導(dǎo)肝癌細(xì)胞EMT及相關(guān)機(jī)制研究[D];中國(guó)人民解放軍醫(yī)學(xué)院;2015年

6 唐俊;miR-612對(duì)肝癌細(xì)胞干性的調(diào)節(jié)作用及其機(jī)制[D];復(fù)旦大學(xué);2014年

7 李韻;miR-27b-5p促進(jìn)肝癌細(xì)胞向肝癌干樣細(xì)胞轉(zhuǎn)化及其機(jī)制研究[D];四川醫(yī)科大學(xué);2015年

8 李霄;BMP4對(duì)肝癌VM形成影響的研究[D];天津醫(yī)科大學(xué);2015年

9 曹漪伊;HBV通過(guò)上調(diào)miR-331-3p的表達(dá)從而促進(jìn)肝癌細(xì)胞的增殖[D];重慶醫(yī)科大學(xué);2015年

10 黃亞運(yùn);以熱休克蛋白為靶點(diǎn)的抗HBV藥物研究[D];湖北工業(yè)大學(xué);2016年

，

本文編號(hào)：1987564