本文關(guān)鍵詞： 多結(jié)節(jié)肝細(xì)胞癌 彌漫型肝細(xì)胞癌 克隆起源 微衛(wèi)星雜合性缺失 肝移植 病理 腫瘤復(fù)發(fā)　出處：《天津醫(yī)科大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：【目的】探討肝細(xì)胞癌肝移植患者鑒定腫瘤克隆起源的臨床及病理意義�！痉椒ā渴占�2005年08月-2010年08月于我院經(jīng)肝移植治療的符合UCSF標(biāo)準(zhǔn)的多結(jié)節(jié)型肝細(xì)胞癌(HCC)患者的病理資料、術(shù)前血清AFP水平及術(shù)后隨訪資料,上述資料不完整的患者予以剔除,共收集患者60例。對60例患者的各自非腫瘤組織及全部腫瘤組織的石蠟包埋塊進(jìn)行切片,切片數(shù)量為5張,厚10μm,常規(guī)二甲苯-乙醇法脫蠟備用。在顯微切割顯微鏡下,精確選擇脫蠟切片的腫瘤及非腫瘤組織,利用石蠟組織DNA提取試劑盒提取腫瘤及非腫瘤組織的基因組DNA。選取12個(gè)高頻肝細(xì)胞癌微衛(wèi)星雜合性缺失(LOH)位點(diǎn),合成引物后采用SSCP-PCR法檢測癌組織發(fā)生LOH的狀態(tài),根據(jù)每例患者不同微衛(wèi)星位點(diǎn)發(fā)生LOH的情況,判定患者的腫瘤克隆起源方式。采用SPSS20.0統(tǒng)計(jì)軟件分析患者腫瘤克隆起源方式與肝移植術(shù)后無瘤生存率、病理學(xué)特征及血清AFP水平之間的相關(guān)性。收集2004年08月-2010年11月于我院經(jīng)肝移植治療的14例彌漫型肝細(xì)胞癌(D-HCC)患者。在14例患者切除的病肝上分別于肝臟左右葉內(nèi)采集腫瘤間隔≥3cm的腫瘤灶各2塊和非腫瘤肝臟組織1塊。各自非腫瘤組織及全部腫瘤組織的石蠟包埋塊進(jìn)行切片,切片數(shù)量為5張,厚10μm,常規(guī)二甲苯-乙醇法脫蠟備用。在顯微切割顯微鏡下,精確選擇脫蠟切片的腫瘤及非腫瘤組織,利用石蠟組織DNA提取試劑盒提取腫瘤及非腫瘤組織的基因組DNA。選取12個(gè)高頻肝細(xì)胞癌微衛(wèi)星雜合性缺失(LOH)位點(diǎn),合成引物后采用SSCP-PCR法檢測癌組織發(fā)生LOH的狀態(tài),根據(jù)每例患者不同微衛(wèi)星位點(diǎn)發(fā)生LOH的情況,判定患者的腫瘤克隆起源方式。收集14例D-HCC患者術(shù)前血清AFP濃度、術(shù)后病肝病理資料、術(shù)后隨訪資料,總結(jié)D-HCC的臨床及病理特征。【結(jié)果】60例多結(jié)節(jié)HCC患者中,男性53例,女性7例;年齡在38歲至71歲之間,平均為53歲。患者肝病背景包括乙肝肝硬化45例、丙肝肝硬化11例、酒精性肝硬化3例、隱源性肝硬化1例。有2個(gè)腫瘤結(jié)節(jié)的患者45例,有3個(gè)腫瘤結(jié)節(jié)患者15例。60例多結(jié)節(jié)HCC患者共采集腫瘤結(jié)節(jié)135個(gè),直徑0.8cm-3.8cm。微衛(wèi)星LOH檢測判定腫瘤克隆起源分型為IM型、IM型+MO型、MO型和不能判定分型4種類型,分別占33.33%(20/60)、8.34%(5/60)、55%(33/60)和3.33%(2/60),腫瘤克隆起源分型不能判定的2例患者剔除,剩余58例患者分為IM組、MO組和IM+MO組,分別占34.48%(20/58)、56.90%(33/58)和8.62%(5/58)。IM組、MO組和IM+MO組的3年累積無瘤生存率、鏡下癌栓發(fā)生率、腫瘤低分化率(Ⅲ級-Ⅳ級)及AFP中位濃度分別為:50.00%、78.79%和40%,100%、18.18%和100%,80%、51.52%和80%,226.80μg/L(2.78μg/L-3000.00μg/L)、24.59μg/L(1.16μg/L-531.30μg/L)和122.58μg/L(16.20μg/L-1055.00μg/L)。IM組和MO組術(shù)前血清AFP水平ROC曲線下面積為0.792,其95%置信區(qū)間(CI)為0.659-0.926,最佳判定界值為122.30μg/L,靈敏度為0.750,特異度為0.818。統(tǒng)計(jì)顯示IM組患者的3年累積無瘤生存率明顯低于MO組患者(P0.05);鏡下癌栓發(fā)生率和術(shù)前血清AFP濃度明顯高于MO組患者(P0.05),IM組患者腫瘤病理分化程度明顯低于MO組患者(P0.05),IM+MO組的無瘤生存率、腫瘤分化程度及鏡下癌栓發(fā)生率均與IM組無明顯差異(P0.05)。14例D-HCC患者的LOH檢測結(jié)果顯示,11例患者腫瘤克隆起源分型為IM,3例患者腫瘤克隆起源分型為MO和IM同時(shí)存在。14例患者術(shù)前血清AFP濃度0.53μg/L-427.04μg/L,9例患者AFP20.00μg/L,其中4例患者AFP200.00μg/L。術(shù)前影像學(xué)檢查顯示,患者肝臟的影像學(xué)表現(xiàn)類似肝硬化,伴肝臟及脾臟不同程度增大,但未發(fā)現(xiàn)占位性病變。腫瘤分布于全肝,直徑0.1-1.0cm,數(shù)量100個(gè),類似肝硬化結(jié)節(jié)。腫瘤病理分級為Ⅰ級-Ⅱ級,所有D-HCC均可檢見鏡下癌栓。肝移植術(shù)后無瘤生存時(shí)間為4.5-37.4個(gè)月,平均為13.5±6.7月�！窘Y(jié)論】1.多結(jié)節(jié)型HCC的腫瘤克隆起源有兩種主要形式:MO型與IM型;MO型在肝移植術(shù)后3年無瘤生存率明顯高于IM型,而混合型(IM+MO型)預(yù)后與IM型相近。腫瘤克隆起源方式或可作為預(yù)測多結(jié)節(jié)HCC肝移植術(shù)后腫瘤復(fù)發(fā)風(fēng)險(xiǎn)的重要參考指標(biāo)。2.綜合腫瘤克隆起源方式、組織病理學(xué)特征及術(shù)前血清AFP水平等腫瘤學(xué)要素,有助于預(yù)測肝移植術(shù)后腫瘤復(fù)發(fā)的風(fēng)險(xiǎn)。3.D-HCC克隆起源分型以IM為主,多預(yù)示廣泛的肝內(nèi)轉(zhuǎn)移;D-HCC發(fā)病隱匿,臨床及影像學(xué)檢查敏感性低,疑診或鑒別困難時(shí),宜借助肝臟組織穿刺活檢明確診斷。
[Abstract]:[Objective] to investigate the clinical and pathological significance of liver transplantation for hepatocellular carcinoma patients with tumor identification of clonal origin. [Methods] multinodular hepatocellular carcinoma from 2005 08 months -2010 years 08 months in our hospital after liver transplantation in the treatment of the UCSF standard (HCC) and pathological data of patients, preoperative serum AFP level and operation after the follow-up data, the data is not complete with to be removed, collected a total of 60 patients. In 60 patients with their non tumor tissue and tumor tissue of paraffin embedded blocks were sliced, cut the number for 5, up to 10 mu m, conventional xylene ethanol method in dewaxing standby. Microdissection under the microscope the precise selection, dewaxed sections of tumor and non tumor tissues using tissue DNA extraction kit to extract tumor and non tumor tissue genomic DNA. from 12 hepatocellular carcinoma high frequency microsatellite loss of heterozygosity (LOH) loci, using synthetic primers after Detection of cancer tissue by SSCP-PCR LOH, according to each patient of different microsatellite loci LOH, determine the patient's tumor clone origin. By using the SPSS20.0 statistical software free survival analysis of patients with tumor clone origin and after liver transplantation, the correlation between the pathological characteristics and the level of serum AFP from 2004. 08 months -2010 year in November in our hospital after liver transplantation in the treatment of 14 cases of diffuse hepatocellular carcinoma (D-HCC) in patients with liver disease. In 14 cases the tumor resection respectively in the liver or tumor interval was 3cm acquisition leaves the 2 and 1 non tumor liver tissue. Their non tumor tissue and all the tumor tissue of paraffin embedded blocks were sliced, cut the number for 5, up to 10 mu m, conventional xylene ethanol method in dewaxing standby. Microdissection under the microscope, accurate selection of dewaxing sections of tumor and non tumor tissue, using Tissue DNA extraction kit to extract tumor and non tumor tissue genomic DNA. from 12 hepatocellular carcinoma high frequency microsatellite loss of heterozygosity (LOH) loci, detection of cancer occur LOH state was synthesized by SSCP-PCR primers, each patient according to different microsatellite loci LOH, determine the patient's tumor clonal origin. 14 D-HCC patients were collected before surgery, postoperative serum AFP concentration, liver pathological data, postoperative follow-up data, summarize the clinical and pathological features of D-HCC. [results] 60 cases of multiple nodules in HCC patients, male 53 cases, female 7 cases; age between 38 to 71 years old, average 53. Patients with liver disease including the background of 45 cases of hepatitis B, hepatitis C cirrhosis in 11 cases, 3 cases of alcoholic liver cirrhosis, 1 cases of cryptogenic cirrhosis. There were 45 cases of patients with 2 nodules, 3 nodules in 15 patients with.60 cases were multiple nodules in patients with HCC were collected and swollen 鐦ょ粨鑺，

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