本文選題：PI3K-Akt通路 + 差異表達(dá)基因　； 參考：《北京協(xié)和醫(yī)學(xué)院》2015年博士論文

【摘要】：第一部分：PI3K-Akt信號通路在肝母細(xì)胞瘤的作用研究背景肝母細(xì)胞瘤(hepatoblastoma, HB)是兒童最常見的肝臟惡性腫瘤。近20年來由于肝腫瘤切除手術(shù)和化療的優(yōu)化結(jié)合,術(shù)前術(shù)后化療方案的個體化措施的發(fā)展,肝母細(xì)胞瘤患兒的預(yù)后有了很大的改善。PI3K-Akt通路是一個經(jīng)典的抗凋亡、促存活的信號轉(zhuǎn)導(dǎo)途徑,在腫瘤發(fā)生、增殖、侵襲、轉(zhuǎn)移中發(fā)揮重要作用。研究目的本研究通過分析PI3K, AKT在肝母細(xì)胞瘤細(xì)胞系及其腫瘤和癌旁組織的表達(dá)情況,了解PI3K-Akt通路在肝母細(xì)胞瘤中的重要意義,為后續(xù)開發(fā)通過抑制該通路治療肝母細(xì)胞瘤的靶向藥物提供理論依據(jù)。研究方法我們通過差異基因表達(dá)篩查發(fā)現(xiàn)PI3K-Akt通路的相關(guān)因子包括PI3K,Akt,PKC,PKB及GSK-3等在腫瘤組織中特異性的高表達(dá)。利用MTT法測定LY294002作用前后肝母細(xì)胞瘤細(xì)胞系HuH-6和HepG2的增殖；通過Western blot分析PI3K通路抑制劑LY294002對肝母細(xì)胞瘤細(xì)胞系中PI3K、Akt表達(dá)的影響；RT-PCR檢測PI3KmRNA在腫瘤和癌旁組織的表達(dá)差異；Western blot檢測PI3K、p-Akt(Ser473)以及總的Akt蛋白在腫瘤和癌旁組織的差異。結(jié)果本研究中,我們發(fā)現(xiàn)HuH-6和HepG2細(xì)胞對不同濃度的PI3K抑制劑LY294002處理具有劑量依懶性增殖抑制；同時,隨著PI3K通路抑制劑LY294002濃度的增加,PI3K、p-Akt(Ser473)蛋白在HuH-6和HepG2細(xì)胞中的表達(dá)逐漸下降,且各組之間的差異存在統(tǒng)計(jì)學(xué)差異(p0.001)；PI3K mRNA在腫瘤組織中的表達(dá)顯著高于癌旁組織；PI3K和p-Akt(Ser473)蛋白在腫瘤組織的表達(dá)顯著高于癌旁組織。而總的Akt和β-actin在兩組病人中無顯著差別。結(jié)論P(yáng)I3K-Akt通路的激活可致使肝母細(xì)胞瘤腫瘤細(xì)胞增殖,相關(guān)基因在腫瘤組織的表達(dá)顯著高于癌旁組織,提示PI3K-Akt通路在肝母細(xì)胞瘤的發(fā)生發(fā)展中起到了重要的作用。第二部分 肝母細(xì)胞瘤預(yù)后危險(xiǎn)因素的評估研究背景肝母細(xì)胞瘤(hepatoblastoma, HB)是兒童最常見的肝臟惡性腫瘤,化療敏感的患兒術(shù)后具有較高的總體生存率(3年生存率在75%以上)。盡管如此,肝母細(xì)胞瘤患兒的預(yù)后仍存在較大差異,因?yàn)槎鄠�危險(xiǎn)因素與其預(yù)后密切相關(guān)。研究目的本研究的目的是確定與亞洲人群肝母細(xì)胞瘤患兒預(yù)后相關(guān)的危險(xiǎn)因素,并分析影響患兒無事件生存(Event-Free Survival, EFS)的各個危險(xiǎn)因素之間的相互關(guān)系。研究方法回顧性分析首都兒科研究所附屬兒童醫(yī)院2001年1月至2014年9月行肝腫瘤切除術(shù)的肝母細(xì)胞瘤患兒共176例,采用Kaplan-Meier生存曲線分析和Cox比例風(fēng)險(xiǎn)模型評估與肝母細(xì)胞瘤相關(guān)的危險(xiǎn)因素。結(jié)果所有患兒(176例)的中位無事件生存時間為80.4個月(95% CI：71.6-89.2)及5年無事件生存率(EFS)54.6%,五年總體生存率(OS)66.7%。肝母細(xì)胞瘤患兒預(yù)后與AFP水平、腫瘤單發(fā)多發(fā)、PRETEXT分期、有無轉(zhuǎn)移、病理分型有關(guān)。多因素分析結(jié)果顯示：AFP水平(HR：2.567,P=0.014),腫瘤是否多發(fā)(HR：2.187,P=0.028),腫瘤是否轉(zhuǎn)移(HR：1.912,P=0.028)以及PRETEXT IV期(HR：2.254,P=0.005)均是影響肝母細(xì)胞瘤患兒EFS的獨(dú)立為危險(xiǎn)因素。不同的病理分型分組相互影響,分別引入多因素Cox模型中進(jìn)行分析提示,完全胎兒型(HR：2.752,P=0.021)是影響肝母細(xì)胞瘤患兒EFS的獨(dú)立為危險(xiǎn)因素。結(jié)論肝母細(xì)胞瘤患兒預(yù)后與AFP水平、腫瘤單發(fā)多發(fā)、PRETEXT分期、有無轉(zhuǎn)移、病理分型有關(guān)。腫瘤的病理分型與其他各因素之間存在著相互作用,不同危險(xiǎn)因素的存在進(jìn)一步影響病理分型對于預(yù)后的預(yù)測。
[Abstract]:The first part: the role of PI3K-Akt signaling pathway in hepatblastoma, background hepatoblastoma (HB) is the most common liver malignant tumor in children. In the last 20 years, the combination of hepatoma resection and chemotherapy, the development of preoperative and postoperative chemotherapy regimens, and the prognosis of the children with hepatblastoma A great improvement in the.PI3K-Akt pathway is a classic anti apoptotic and survival signal transduction pathway that plays an important role in tumor genesis, proliferation, invasion and metastasis. The purpose of this study was to analyze the expression of PI3K, AKT in the hepatblastoma cell lines and their tumor and paracancerous groups, and to understand the PI3K-Akt pathway in the hepatocytes. The important significance of the tumor is to provide a theoretical basis for subsequent development of targeted drugs for the treatment of hepatblastoma by inhibiting this pathway. We found that the related factors of PI3K-Akt pathway include PI3K, Akt, PKC, PKB and GSK-3 in specific high expression in tumor tissues by differential gene expression screening. The determination of LY294002 by MTT method The proliferation of hepatblastoma cell lines HuH-6 and HepG2, and the effect of LY294002 on the expression of PI3K and Akt in the hepatblastoma cell lines by Western blot; RT-PCR detected by RT-PCR to detect the difference in the expression of PI3KmRNA in the tumor and adjacent tissues; Western blot, and the total protein in tumor and cancer In this study, we found that HuH-6 and HepG2 cells have a dose dependent inhibition of laziness in different concentrations of PI3K inhibitor LY294002; meanwhile, the expression of PI3K, p-Akt (Ser473) protein in HuH-6 and HepG2 cells gradually decreases with the increase of LY294002 concentration in the PI3K pathway, and the difference between each group is poor. The difference was statistically significant (p0.001), and the expression of PI3K mRNA in the tumor tissues was significantly higher than that in the para cancerous tissue, and the expression of PI3K and p-Akt (Ser473) protein in the tumor tissues was significantly higher than that in the paracancerous tissues. The total Akt and beta -actin were not significantly different in the two groups. Conclusion the activation of PI3K-Akt path can lead to the proliferation of hepatblastoma tumor cells. The expression of related genes in tumor tissues is significantly higher than that of paracancerous tissue, suggesting that PI3K-Akt pathway plays an important role in the development of hepatblastoma. Second the evaluation of prognostic risk factors in partial hepatblastoma (hepatoblastoma, HB) is the most common malignant tumor of the liver in children, chemotherapy sensitivity A higher overall survival rate (3 year survival rate is above 75%) after surgery. However, there are still significant differences in the prognosis of children with hepatblastoma, because multiple risk factors are closely related to their prognosis. The purpose of this study is to determine the risk factors associated with the prognosis of children with Asians with hepatblastoma, and the purpose of this study is to determine the risk factors for the prognosis of the children with hepatblastoma. A retrospective analysis of 176 children with hepatoblastoma in children's Hospital Affiliated to the capital Pediatrics Institute from January 2001 to September 2014, using Kaplan-Meier survival curve analysis and Cox proportions risk, was analyzed in a retrospective analysis of the relationship between the various risk factors affecting Event-Free Survival (EFS). The model assessed the risk factors associated with hepatblastoma. Results the median survival time of all children (176 cases) was 80.4 months (95% CI:71.6-89.2) and 5 years of event free survival (EFS) 54.6%. The prognosis of the five year total survival rate (OS) 66.7%. hepatblastoma was associated with the AFP level, the multiple tumors, the PRETEXT staging, the metastasis, and the disease. The results of multi factor analysis showed that AFP level (HR:2.567, P=0.014), tumor multiple (HR:2.187, P=0.028), tumor metastasis (HR:1.912, P=0.028), and PRETEXT IV phase (HR:2.254, P=0.005) were all risk factors affecting the independence of EFS in children with hepatblastoma. Different pathological classification groups influenced each other, respectively. In the multi factor Cox model, it is suggested that complete fetal type (HR:2.752, P=0.021) is an independent risk factor affecting EFS in children with hepatblastoma. Conclusion the prognosis of the children with hepatblastoma is related to the level of AFP, the multiple tumors, the PRETEXT staging, the metastasis and the pathological classification. The pathological classification of the tumor is associated with the other factors. The presence of interaction and different risk factors further affect the prognosis of pathological classification.

【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R735.7

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 黃秀蘭;崔國輝;周克元;;PI3K-Akt信號通路與腫瘤細(xì)胞凋亡關(guān)系的研究進(jìn)展[J];癌癥;2008年03期

2 魏艇;楊體泉;羅意革;董淳強(qiáng);;14例兒童肝母細(xì)胞瘤綜合治療的療效觀察[J];中國當(dāng)代兒科雜志;2009年06期

3 陳干農(nóng);馬勇;楊竹林;;兒童肝母細(xì)胞瘤肝組織中GRP78和GRP94表達(dá)及其臨床病理意義[J];中國當(dāng)代兒科雜志;2010年08期

4 李明皓;嚴(yán)律南;王樹人;;改良式全肝血流阻斷包膜外切除肝巨大血管瘤[J];南方醫(yī)科大學(xué)學(xué)報(bào);2007年06期

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本文編號：1867825