【摘要】：[目的]膽囊癌是一類具有高度惡性、高致死性、高侵襲性等特點的消化道惡性腫瘤,因發(fā)病隱匿、無特異臨床表現(xiàn)導(dǎo)致其面臨早診困難,且無有效治療方案。膽囊癌患者絕大多數(shù)被確診時已是晚期,失去手術(shù)治愈的機會。而膽囊癌的發(fā)病機制尚不清楚,亟需探索相關(guān)病理機制和有效的診療策略。本課題旨在從膽囊癌與癌旁正常組織的差異表達基因中,尋找候選癌基因,并確定其在膽囊癌變多階段中與臨床病理參數(shù)聯(lián)系的臨床意義,初步探索該基因在膽囊癌發(fā)生發(fā)展中的作用,以期為膽囊癌患者的診治提供新的研究線索。[方法]我們采用Agilent mRNA表達譜芯片技術(shù)對膽囊癌和配對的癌旁正常組織進行差異表達基因的mRNA表達譜分析;通過分子功能聯(lián)合通路的表達關(guān)聯(lián)分析篩選獲得目的研究基因;進而采用免疫組織化學(xué)技術(shù)對比檢測其在慢性膽囊炎、癌前病變和膽囊癌組織中的表達情況;利用SPSS19.0軟件統(tǒng)計分析其與膽囊癌變多階段、臨床病理參數(shù)的相關(guān)性。[結(jié)果]1.通過Agilent mRNA表達譜芯片篩選提示OLFM4等379個基因在膽囊癌組織中高表達。利用Agilent mRNA表達譜芯片(SurePrint G3 Human Gene Expression 8x60K v2 Microarray)篩選技術(shù),我們在5對膽囊癌及癌旁正常組織中,篩選出631個差異表達的mRNA。其中,上調(diào)表達基因379個,如OLFM4、CDH3、STYXL1 等;下調(diào)表達基因 252 個,如 YKT6、LUZP1、FBLN1 等;2. OLFM4基因在膽囊臨床組織樣本中蛋白細胞定位隨癌變階段發(fā)展從胞核轉(zhuǎn)移至胞外,且隨癌變階段發(fā)展表達率和強度逐漸增高。2.1 OLFM4在膽囊癌變多階段發(fā)展進程中細胞定位存在從胞核到胞外的轉(zhuǎn)移現(xiàn)象。OLFM4在慢性膽囊炎組織中主要表達于細胞核,少量表達于細胞質(zhì),在癌前病變組織中,主要表達于細胞質(zhì)和胞外,在膽囊癌組織中,主要分泌到細胞外;2.2 OLFM4表達率在癌變多階段中表達逐步增加。OLFM4表達率在膽囊癌癌變多階段(慢性膽囊炎,癌前病變,膽囊癌)組織中表達率逐步增加,且差異具有顯著性(41.67% vs71.74%vs 80.56%,p0.001)。其中,OLFM4在癌前病變中表達高于慢性膽囊炎,差異具有顯著性(p0. 001) ; OLFM4在膽囊癌中高表達于癌前病變,但差異未見顯著性(p=0.192) ; OLFM4在膽囊癌中表達高于慢性膽囊炎,且差異具有顯著性(p0. 001);2.3 OLFM4表達強度在膽囊癌變多階段組織中逐步增強。 OLFM4表達強度在慢性膽囊炎、癌前病變和膽囊癌中依次增強,差異具有顯著性(105.90vs 149.33 vs 166.01,p0.001)。其中,OLFM4在膽囊癌組織中的表達強度強于慢性膽囊炎組織,差異具有顯著性(p0.001) ; OLFM4在膽囊癌前病變組織中的表達強度強于慢性膽囊炎組織,差異具有顯著性(p0.001); OLFM4在膽囊癌中的表達強度強于癌前病變組織,但差異未見顯著性(p=0.123);2.4 OLFM4表達水平與其他臨床病理參數(shù)未見顯著相關(guān)性。在膽囊癌臨床組織樣本中,OLFM4表達水平與患者年齡、性別、T分期、N分期、臨床分期、分化程度均未見顯著相關(guān)性(p 0.05)。[結(jié)論]1. OLFM4在膽囊癌變多階段發(fā)展進程中可能發(fā)揮不同的作用,具有階段特異性;2. OLFM4可能促進慢性膽囊炎-癌前病變-膽囊癌發(fā)展進程。
[Abstract]:[Objective] Gallbladder cancer is a kind of digestive tract malignant tumor with high degree of malignancy, high lethal and high invasiveness. The majority of the patients with gallbladder cancer have been diagnosed with an advanced stage and have no chance of surgical treatment. The pathogenesis of gallbladder carcinoma is not clear, and it is urgent to explore the relevant pathological mechanism and effective diagnosis and treatment strategy. The purpose of this study is to find the candidate oncogenes from the differential expression genes of the normal tissues of the gallbladder carcinoma and the adjacent tissues, and to determine the clinical significance of their association with the clinical and pathological parameters in the multi-stage gallbladder carcinoma, and to explore the role of the gene in the development of the gallbladder carcinoma. In order to provide a new clue for the diagnosis and treatment of the gallbladder carcinoma. [Methods] The expression profile of the differentially expressed genes in the normal tissues of the gallbladder and the adjacent non-adjacent normal tissues was analyzed by using the Agilent mRNA expression profile chip technique, and the target study gene was screened by the expression association analysis of the molecular function combined pathway. Furthermore, the expression of its expression in the tissues of chronic cholecystitis, pre-cancerous and gallbladder carcinoma was detected by immunohistochemical technique, and the correlation between the multi-stage and the clinicopathological parameters of the gallbladder carcinoma was statistically analyzed by using the SPSS19.0 software. [Results] 1. 379 genes, such as OLFM4, were expressed in the gallbladder carcinoma tissue by an Agilent mRNA expression profile. By using the Agilent mRNA expression spectrum chip (SurePrint G3 Human Gene Expression 8x60K v2 Microarray), we screened 631 differentially expressed mRNA in the normal tissues of the gallbladder carcinoma and the adjacent to the cancer. Of these,379 are up-regulated, such as OLFM4, CDH3, STYXL1, etc., and 252 down-regulated genes, such as YKT6, LUZP1, FBLN1, etc.; The localization of the OLFM4 gene in the sample of the gallbladder clinical tissue has been transferred from the nucleus to the extracellular domain with the development of the carcinogenesis stage, and the expression rate and the intensity are gradually increased with the development of the carcinogenesis stage. OLFM4 is mainly expressed in the nucleus of the chronic cholecystitis and is expressed in the cytoplasm, which is mainly expressed in the cytoplasm and the extracellular in the tissues of the precancerous lesion, and is mainly secreted outside the cell in the gallbladder carcinoma tissue, and the expression rate of the OLFM4 is gradually increased in the multi-stage of the cancer. The expression rate of OLFM4 was increased gradually in the multiple stage of gallbladder carcinoma (chronic cholecystitis, precancerous lesion and gallbladder carcinoma), and the difference was significant (41.67% vs.74% vs. 80.56%, p0.001). Among them, the expression of OLFM4 in the pre-cancerous lesions was higher than that of chronic cholecystitis and the difference was significant (p0. There was no significant difference in the expression of OLFM4 in the gallbladder carcinoma (p = 0.192), and the expression of OLFM4 in the gallbladder carcinoma was higher than that of the chronic cholecystitis, and the difference was significant (p0. 001); 2.3 OLFM4 expression intensity was gradually increased in the multi-stage tissue of gallbladder carcinoma. The expression intensity of OLFM4 was significantly increased in chronic cholecystitis, pre-cancerous and gallbladder carcinoma (105.90 vs. 149.33 vs. 166.01, p0.001). The expression of OLFM4 in the gallbladder carcinoma was stronger than that of the chronic cholecystitis (p0.001), and the expression of OLFM4 in the premalignant tissue of the gallbladder was stronger than that of the chronic cholecystitis (p0.001). The expression of OLFM4 in gallbladder carcinoma was stronger than that of precancerous lesion, but the difference was not significant (p = 0.123), and the expression level of 2.4 OLFM4 was not significantly correlated with other clinical pathological parameters. In the sample of gallbladder carcinoma, the expression level of OLFM4 was not significantly correlated with age, sex, T stage, N stage, clinical stage and degree of differentiation (p.05). [Conclusion] 1. OLFM4 may play a different role in the development of the multi-stage of gallbladder carcinoma, and has stage specificity;2. OLFM4 may promote the progression of chronic cholecystitis-premalignant lesions-the development of the gallbladder carcinoma.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.8

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相關(guān)期刊論文 前1條

1 朱凱;張愛萍;林秀敏;;非小細胞肺癌患者血漿OLFM4水平及其臨床意義分析[J];臨床腫瘤學(xué)雜志;2016年06期

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本文編號：2469156