本文選題：肝細胞肝癌　切入點：COX-2　出處：《新鄉(xiāng)醫(yī)學院》2015年碩士論文　論文類型：學位論文

【摘要】：目的目前,癌癥已經(jīng)成為人類健康的第一殺手,尤其是肝細胞肝癌(Hepatocellular carcinoma, HCC),在中國甚至世界范圍內(nèi),發(fā)病率和死亡率在逐年升高。因此,尋找新的診斷和治療相關分子協(xié)助臨床醫(yī)生快速有效地診斷和治療HCC將有利于提高HCC病人的療效和生存率。環(huán)氧合酶(cyclooxygenase, COX)在體內(nèi)有著重要的作用,近年來的研究表明,Ⅱ型環(huán)氧合酶(COX-2)在食管癌、肝癌中表達增高,與腫瘤血管的形成密切相關。CD88又稱補體5片段受體(complement component 5a receptor, C5aR),常被用來作為鑒定白細胞系的標志,與C5a結合后能夠激活PI3K/A KT/mTOR和IL-6/STAT3兩個信號通路并促進肝細胞的分裂增殖和對抗凋亡,趨化白細胞和參與炎癥反應,在腫瘤侵襲轉移中發(fā)揮重要作用。已有研究顯示CD88可能促進腫瘤周圍血管新生,加速腫瘤細胞侵襲轉移。由于COX-2和CD88在腫瘤組織中的表達均顯著增高,并且與腫瘤的侵襲轉移密切相關,因此,本研究將探討二者在肝細胞肝癌組織中的表達率以及二者之間的相關性。方法1.標本收集收集新鄉(xiāng)市第一人民醫(yī)院病理科2013年1月至2014年4月期間,所有資料完整的肝細胞肝癌組織共57例。根據(jù)年齡、性別、腫瘤直徑、分化程度、分期和有無淋巴結轉移等將標本分類。一部分組織放入液氮保存,另一部分組織經(jīng)10%的福爾馬林溶液固定、石蠟包埋以及4gm厚度連續(xù)切片。2. RT-PCR檢測正常肝組織和肝細胞肝癌組織中COX-2和CD88mRNA的表達提取肝正常組織和肝細胞肝細胞肝癌組織中總mRNA,檢測組織中COX-2和CD88 mRNA的表達情況。3.免疫組織化學染色觀察COX-2和CD88蛋白的表達將準備好的組織切片用SABC法進行免疫組織化學染色,觀察并確定COX-2和CD88的表達部位,計算二者的表達率以及表達的相關性。4.結果判斷RT-PCR的結果用Image J軟件測量灰度值,根據(jù)與內(nèi)參的對比,判斷COX-2和CD88 mRNA的表達。免疫組織化學結果根據(jù)染色強度和陽性細胞所占的百分比分別計分,再將染色強度分值與陽性細胞所占百分比分值相乘所得乘積判定染色結果。5.統(tǒng)計學處理計量數(shù)據(jù)均使用均數(shù)±標準差(x±s)表示,應用SPSS 17.0統(tǒng)計軟件對數(shù)據(jù)進行統(tǒng)計學處理。多樣本均數(shù)的比較采用方差分析；陽性率的比較用χ2檢驗；相關性分析用Pearson檢驗,檢驗水準均設為α=0.05,即P0.05有顯著統(tǒng)計學差異,P0.01有非常顯著統(tǒng)計學差異,P0.05無統(tǒng)計學差異。結果1.COX-2和CD88 mRNA的表達COX-2和CD88 mRNA在正常肝組織和肝細胞肝癌組織中均有表達,且在肝細胞肝癌組織的表達較正常肝組織明顯增高。二者在低、中、高分化癌組織中表達逐漸增高,低、中分化組表達無統(tǒng)計學差異,而二者與高分化組相比有統(tǒng)計學差異(P0.05)。2.COX-2在肝細胞癌中的表達及與臨床病理特征的關系COX-2染色陽性的細胞其細胞漿中呈現(xiàn)彌漫性分布的黃色和棕黃色顆粒狀物質(zhì),肝細胞癌組織中的COX-2陽性率為73.68%。COX-2蛋白的表達與腫瘤分化程度、腫瘤大小、臨床分期、淋巴結轉移情況無關(P0.05)。3.CD88在肝細胞癌中的表達及與臨床病理特征的關系CD88蛋白表達主要位于細胞膜,部分位于胞漿中。肝細胞癌組織中的CD88蛋白表達的陽性率為82.45%。CD88蛋白的表達與腫瘤分化程度、腫瘤大小、臨床分期、淋巴結轉移情況無關(P0.05)3.COX-2與CD88在肝細胞癌中的表達的相關性肝癌組織中,COX-2陽性組,CD88的陽性率為90.48%,顯著高于COX-2陰性組(60%),COX-2和CD88的蛋白表達呈正相關(r=0.353,P0.05)。結論1.肝細胞肝癌組織中COX-2和CD88 mRNA的表達較正常肝組織明顯增多,但COX-2和CD88蛋白的表達與腫瘤分化程度、腫瘤大小、臨床分期和淋巴結轉移均無關。2.肝細胞肝癌組織中COX-2和CD88蛋白的表達呈正相關。
[Abstract]:At present, cancer has become the first killer of human health, especially hepatocellular carcinoma (Hepatocellular, carcinoma, HCC) in China even in the world, the morbidity and mortality increased year by year. Therefore, finding new molecular diagnosis and treatment to assist clinicians effectively in diagnosis and treatment of HCC will improve HCC patients curative effect and survival rate. Cyclooxygenase (cyclooxygenase, COX) plays an important role in the body, recent studies show that cyclooxygenase-2 (COX-2) expression in esophageal cancer, liver cancer, and tumor angiogenesis is closely related to the.CD88 also known as complement 5 receptor (complement component fragment 5A receptor, C5aR), marks are often used as identification of white blood cells, when combined with C5a can activate PI3K/A KT/mTOR and IL-6/STAT3 two signaling pathway and promote liver cell proliferation and apoptosis resistance Dead, chemotaxis of white blood cells and participate in inflammation, play an important role in tumor invasion and metastasis. Studies have shown that CD88 may promote tumor angiogenesis, accelerate tumor cells metastasis. The expression of COX-2 and CD88 in tumor tissues were significantly increased, and the tumor invasion and metastasis are closely related, therefore, the the research will explore the correlation between the expression rate of two in hepatocellular carcinoma and two. Methods: 1. tissue samples collected from Xinxiang First People's Hospital Department of pathology from January 2013 to April 2014, all data of hepatocellular carcinoma in 57 cases. According to age, gender, tumor size, degree of differentiation, staging and lymph node metastasis as part of the organization classification. Specimens were preserved in liquid nitrogen, the other part of the organization in 10% formalin fixed, paraffin embedded and 4gm continuous.2. R slice thickness The expression of COX-2 and CD88mRNA T-PCR detection of normal liver tissue and hepatocellular carcinoma total mRNA was extracted from normal liver tissue and liver cells in hepatocellular carcinoma, the expression of.3. COX-2 and CD88 immunohistochemical staining in detection of mRNA expression of COX-2 and CD88 proteins will be prepared by the method of SABC immune tissue histochemical staining, observe and determine the expression site of COX-2 and CD88. The correlation between the expression of.4. two and the expression rate of calculation results to determine the results of RT-PCR using Image J software to measure the gray value, according to the comparison and reference, the expression of COX-2 and CD88 mRNA found fault. According to the results of immunohistochemical staining intensity and the percentage of positive cells for the score, the staining intensity score and the percentage of the positive cells were obtained by multiplying the product to determine the staining results statistically.5. measurement data are made With the mean and standard deviation (x + s), using SPSS 17 statistical software for statistical processing of data. Compared with the mean diversity analysis of variance; the positive rate of the 2 test was used to compare; correlation analysis with Pearson test, the test level for alpha =0.05, P0.05 had significant difference P0.01, there are very significant differences, no significant difference P0.05. Results the expression of COX-2 and CD88 mRNA 1.COX-2 and CD88 mRNA in normal liver tissue and hepatocellular carcinoma tissues, and the expression in hepatocellular carcinoma tissues than in normal liver tissue were significantly increased. Two in the low and high differentiation cancer tissues the expression gradually increased in low differentiation group, there were no significant difference, but the two compared with the high differentiation group had significant difference (P0.05) expression of.2.COX-2 in hepatocellular carcinoma and its relationship with clinical pathological characteristics of COX-2 positive cells The cytoplasm showed diffuse distribution of yellow and brown granular material, the positive rate of COX-2 in hepatocellular carcinoma 73.68%.COX-2 protein expression and tumor differentiation, tumor size, clinical staging, independent of lymph node metastasis (P0.05) expression of.3.CD88 in hepatocellular carcinoma and its relationship with clinical pathological characteristics CD88 protein was mainly located in the cell membrane, located in the cytoplasm. The positive expression rate of hepatocellular carcinoma CD88 protein 82.45%.CD88 protein expression and tumor differentiation, tumor size, clinical stage, lymph node metastasis (P0.05). The expression of 3.COX-2 and CD88 in hepatocellular carcinoma associated hepatocellular carcinoma in COX-2 positive group, the positive rate of CD88 was 90.48%, significantly higher than that of COX-2 negative group (60%), COX-2 and CD88 protein expression were positively correlated (r=0.353, P0.05) COX-2 and CD 1.. Conclusion liver cells in hepatocellular carcinoma 88 mRNA expression increased significantly compared with normal liver tissue, but the expression of COX-2 and CD88 protein was not related to tumor differentiation, tumor size, clinical stage and lymph node metastasis. The expression of COX-2 and CD88 protein in.2. HCC tissues was positively correlated.

【學位授予單位】：新鄉(xiāng)醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.7

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