本文選題：胰腺 + 實(shí)性假乳頭狀腫瘤��； 參考：《北京協(xié)和醫(yī)學(xué)院》2016年博士論文

【摘要】：胰腺實(shí)性假乳頭狀瘤(pancreatic solid pseudopapillary neoplasm, SPN)是一種少見的具有惡性潛能的胰腺腫瘤。胰腺SPN發(fā)病率低,僅占胰腺腫瘤的1%-2%,多見于年輕女性。腫瘤起病隱匿,大部分患者無臨床癥狀,有癥狀的患者表現(xiàn)為腹部不適,惡心,嘔吐等,位于胰頭的SPN偶見黃疸。由于對(duì)此腫瘤的認(rèn)識(shí)不斷加深和影像學(xué)技術(shù)的改進(jìn),胰腺SPN的檢出率在逐漸增加。胰腺SPN臨床少見,具有獨(dú)特的臨床表現(xiàn)和生物學(xué)行為,然而目前相關(guān)基礎(chǔ)及臨床研究較少�；蛲蛔儥z測(cè)目前已廣泛應(yīng)用于腫瘤的相關(guān)研究,對(duì)于揭示腫瘤的發(fā)生發(fā)展過程起到了重要作用。新一代測(cè)序技術(shù)(next-generation sequencing technology, NGS)以高通量測(cè)序?yàn)樘攸c(diǎn),包括全基因組測(cè)序和外顯子測(cè)序。外顯子測(cè)序是指利用目標(biāo)序列捕獲技術(shù)將基因組的全部外顯子區(qū)域DNA捕獲后并通過NGS技術(shù)進(jìn)行測(cè)序的方法。其測(cè)序過程稱為“循環(huán)芯片測(cè)序法”,是對(duì)布滿DNA樣品的芯片重復(fù)進(jìn)行基于DNA的聚合酶反應(yīng)(模板變性、引物退火雜交及延伸)以及熒光序列讀取。外顯子測(cè)序針對(duì)性強(qiáng),覆蓋度深,數(shù)據(jù)準(zhǔn)確性高,具有簡(jiǎn)便、經(jīng)濟(jì)、高效等優(yōu)點(diǎn),已經(jīng)被廣泛應(yīng)用于腫瘤疾病的研究中。本研究對(duì)胰腺SPN新鮮組織樣本提取DNA,構(gòu)建DNA文庫(kù)并進(jìn)行高通量外顯子測(cè)序,初步篩選出突變基因共94個(gè)。進(jìn)一步對(duì)突變基因進(jìn)行生物信息學(xué)分析和Sanger測(cè)序驗(yàn)證,證實(shí)CTNNB1是胰腺SPN的顯著突變基因。另對(duì)12個(gè)胰腺SPN腫瘤組織進(jìn)行擴(kuò)大樣本PCR驗(yàn)證,結(jié)果表明CTNNB1的突變率為93.3%(14／15)。針對(duì)CTNNB1編碼的β-catenin蛋白共進(jìn)行了91例石蠟組織切片的免疫組織化學(xué)染色,共獲得有效染色87例,結(jié)果顯示共有72例組織切片為陽性,陽性率為82.8%(72／87)。相關(guān)性分析表明β-catenin免疫組化陽性表達(dá)程度與腫瘤直徑有相關(guān)性(P=0.029)。此外,本研究回顧性分析2000至2010年之間所有接受手術(shù)并具有完整臨床病理資料的病例。共入組100例病人,其中84例為女性,16例為男性,中位年齡31(13-68)歲。腫瘤平均直徑6.5(1.5-18)cm。共有24例患者診斷為惡性胰腺SPN。49例患者接受了淋巴結(jié)清掃,術(shù)后病理提示僅有4例患者有淋巴結(jié)轉(zhuǎn)移。單因素分析表明,術(shù)前CT/MRI顯示胰周淋巴結(jié)腫大是惡性胰腺SPN的危險(xiǎn)因素(P=0.025)。遠(yuǎn)期隨訪結(jié)果顯示,共有兩名患者術(shù)后出現(xiàn)了肝轉(zhuǎn)移,并接受了肝轉(zhuǎn)移瘤切除術(shù)。兩名患者分別隨訪24和32個(gè)月,未出現(xiàn)腫瘤復(fù)發(fā)。綜上所述,本論文對(duì)胰腺SPN的分子機(jī)制和臨床病理特征進(jìn)行了一系列研究,表明CTNNB1基因突變?cè)谝认賁PN發(fā)生發(fā)展過程中起到重要作用,說明β-catenin表達(dá)與胰腺SPN腫瘤增殖密切相關(guān),有希望成為胰腺SPN診斷和治療的分子靶點(diǎn)。此外,本研究還證明了術(shù)前影像學(xué)胰周淋巴結(jié)腫大與惡性胰腺SPN相關(guān)。長(zhǎng)期隨訪結(jié)果提示,對(duì)惡性胰腺SPN患者應(yīng)進(jìn)行密切隨訪和定期復(fù)查。
[Abstract]:Pancreatic solid pseudopapillary neoplasm (SPNs) is a rare pancreatic tumor with malignant potential. The incidence of pancreatic SPN was low, accounting for only 1-2 percent of pancreatic tumors, and was more common in young women. Most of the patients had no clinical symptoms. The symptomatic patients showed abdominal discomfort, nausea, vomiting, etc. The SPN located in the head of pancreas occasionally showed jaundice. The detection rate of pancreatic SPN is gradually increasing due to the deepening understanding of the tumor and the improvement of imaging techniques. Pancreatic SPN is rare and has unique clinical manifestations and biological behaviors. However, there are few basic and clinical studies. Gene mutation detection has been widely used in tumor related research, which plays an important role in revealing the process of tumor occurrence and development. The next-generation sequencing technology is characterized by high throughput sequencing, including whole genome sequencing and exon sequencing. Exon sequencing is a method which uses target sequence capture technology to capture all exon regions of genomic DNA and sequenced by NGS technology. The sequencing process is called "cyclic chip sequencing", which repeats the microarray covered with DNA samples by DNA based polymerase reaction (template denaturation, primer annealing hybridization and extension) and fluorescence sequence reading. Exon sequencing has been widely used in the study of tumor diseases due to its strong pertinence, deep coverage, high accuracy, simple, economical and efficient. In this study, DNA library was constructed from fresh tissue samples of pancreatic SPN, and high throughput exon sequencing was carried out. A total of 94 mutant genes were screened. Further bioinformatics analysis and Sanger sequencing confirmed that CTNNB1 was a significant mutant gene of pancreatic SPN. In addition, 12 pancreatic SPN tumor tissues were tested by PCR. The results showed that the mutation rate of CTNNB1 was 93. 3% and 14 / 15%. A total of 91 cases of paraffin sections were stained by immunohistochemistry for 尾 -catenin protein encoded by CTNNB1. 87 cases were stained effectively. The results showed that 72 cases were positive, and the positive rate was 82.8%. Correlation analysis showed that the expression of 尾 -catenin was correlated with the diameter of tumor. In addition, this study retrospectively analyzed all cases with complete clinicopathologic data between 2000 and 2010. A total of 100 patients were enrolled, of whom 84 were female and 16 were male, with a median age of 31 13-68 years. The mean diameter of tumor was 6.5 ~ 1.5-18 cm. A total of 24 patients were diagnosed as malignant pancreatic SPN.49 patients underwent lymph node dissection, and only 4 patients had lymph node metastasis after operation. Univariate analysis showed that preoperative CT/MRI showed peripancreatic lymphadenopathy as a risk factor for malignant pancreatic SPN. Long-term follow-up showed that two patients had postoperative liver metastases and underwent resection of liver metastases. The two patients were followed up for 24 and 32 months, respectively, without recurrence. In conclusion, a series of studies on the molecular mechanism and clinicopathological characteristics of pancreatic SPN have been carried out in this paper, which indicate that CTNNB1 gene mutation plays an important role in the pathogenesis and development of pancreatic SPN, suggesting that 尾 -catenin expression is closely related to the proliferation of pancreatic SPN tumors. It is promising to be a molecular target for the diagnosis and treatment of pancreatic SPN. In addition, this study also demonstrated that preoperative imaging peripancreatic lymphadenopathy was associated with malignant pancreatic SPN. Long-term follow-up results suggest that patients with malignant pancreatic SPN should be closely followed-up and regularly rechecked.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R735.9

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