本文選題：結腸炎相關結腸癌 + PLK1　； 參考：《北京協(xié)和醫(yī)學院》2017年碩士論文

【摘要】：結直腸癌(Colorectalcancer,CRC)是最常見的消化道腫瘤之一。近年,我國結直腸癌的發(fā)病率呈逐年上升趨勢。炎性腸病(inflammatory bowel disease,IBD)作為一種反復發(fā)作的慢性炎癥,是結腸癌發(fā)生的重要高危因素之一,約20%的結腸炎患者最終會發(fā)展為結腸炎相關結腸癌(Colitis-associated Cancer,CAC)。結腸炎向結腸癌轉化是一個涉及到多個信號通路和分子的復雜過程,目前相關的機制尚不明確。絲氨酸/蘇氨酸蛋白激酶PLKl(polo-likekinasel)是哺乳動物有絲分裂進程中的關鍵調節(jié)因子。大量研究結果顯示PLK1在包括結直腸癌(colorectal cancer,CRC)在內的多種惡性腫瘤組織中表達升高,但迄今為止PLK1與CAC之間的關系尚無文獻報道。因此本研究旨在探索PLK1在炎癥、免疫及結腸癌之間的潛在聯(lián)系并初步探討PLK1在CAC發(fā)生的作用及相關分子機制。為探索PLK1與CAC發(fā)生之間的關系,本研究利用氧化偶氮甲烷(azoxymethane,AOM)和葡聚糖硫酸鈉(dextran sulfate sodium,DSS)誘導 PLK1基因敲入小鼠(PLK1fl/+小鼠,實驗組)及同窩野生型小鼠(WT小鼠,對照組)發(fā)生CAC,結果顯示PLK1fl/+小鼠結腸腫瘤數量和腫瘤負荷顯著低于WT對照小鼠,提示PLK1過表達可有效抑制AOM/DSS誘導的CAC的發(fā)生。以往研究發(fā)現PLK1在多種實體腫瘤中發(fā)揮促癌作用,因此亟需揭示PLK1過表達抑制CAC發(fā)生的作用機制。我們使用免疫組織化學(IHC)染色法檢測了AOM/DSS誘導野生型小鼠形成的CAC病灶部位中PLK1的表達情況,發(fā)現其在腸道上皮間質細胞中有高水平表達。據此,我們在小鼠結腸炎相關結腸癌腫瘤組織的連續(xù)切片中進行了 PLK1和Gr1(標記髓系來源細胞)的IHC染色,發(fā)現PLK1和Gr1陽性細胞在結腸病灶組織中有一致的定位。免疫熒光(IF)共定位檢測結果顯示,PLK1與Gr1標記的髓系細胞及F4/80標記的巨噬細胞存在共定位現象。上述結果顯示CAC發(fā)生過程中PLK1主要表達于腸道炎癥微環(huán)境中的髓系細胞。腸炎/腸癌微環(huán)境中的巨噬細胞等髓系細胞可通過分泌促炎細胞因子及趨化因子促進結腸炎向結腸癌的轉化。為進一步揭示PLK1在腸道炎癥微環(huán)境髓系細胞中表達抑制CAC發(fā)生的分子機制,我們使用細胞因子芯片檢測經DSS處理的PLK1fl/+和WT小鼠的結腸組織培養(yǎng)上清中炎性因子的表達情況。芯片檢測結果顯示PLK1高表達可以明顯抑制小鼠結腸病灶組織表達和分泌包括TNF-α在內的多個細胞因子和趨化因子。ELISA和qRT-PCR檢測證實了PLK1fl/+小鼠結腸組織中TNF-α表達和分泌的改變。我們進一步使用PLK1特異性的siRNA或小分子抑制劑處理體外培養(yǎng)的巨噬細胞J774.1,發(fā)現抑制PLK1的表達可明顯上調TNF-α表達。上述結果提示,PLK1可能通過抑制腸道炎癥微環(huán)境中髓系細胞的促炎和趨化功能進而抑制CAC的發(fā)生。綜上所述,本研究結果提示PLK1可能參與調控間質髓系細胞的促炎/促癌功能,其可能通過抑制TNF-α等炎性因子的表達和分泌進而抑制AOM/DSS誘導的CAC的發(fā)生。本研究不僅有助于闡明PLK1在腸炎/腸癌相關免疫細胞中新的作用及分子機制,而且可能為今后開發(fā)針對免疫微環(huán)境的治療策略提供理論依據和實驗基礎。結直腸癌作為常見惡性腫瘤,極大的威脅著人類的健康和生命。結直腸癌術后復發(fā)轉移是導致患者死亡的主要原因之一,因此尋找能夠用于預測結直腸癌復發(fā)轉移或準確判斷預后的分子標志物對于提高結直腸癌患者的生存率具有重要意義。本實驗室在前期工作中發(fā)現KIAA1522在食管鱗狀細胞癌、肺癌等腫瘤中異常表達,并與患者的預后相關。但迄今為止,KIAA1522在結直腸癌中的表達改變尚不清楚。本研究應用組織芯片-免疫組織化學染色技術,對96例結直腸癌組織及其配對癌旁正常組織中KIAA1522蛋白的表達情況進行檢測,并對其表達改變與結直腸癌臨床病理指標及患者預后的關系進行統(tǒng)計分析。免疫組織化學分析結果顯示結直腸上皮細胞中KIAA1522蛋白主要定位于細胞漿。在結直腸癌組織中,KIAA1522蛋白表達的陽性率為81%(78/96);而在配對的癌旁正常組織中其表達的陽性率為13%(12/96),兩者間差異顯著(P0.05)。進一步的統(tǒng)計分析結果顯示,KIAA1522蛋白過表達與結直腸癌患者術后三年無瘤生存期短顯著正相關(P = 0.017),并且與遠處轉移正相關(P = 0.012)。多因素Cox回歸分析結果表明,KIAA1522過表達是結直腸癌患者預后不良的獨立預測因素(P= 0.020)。綜上所述,KIAA1522在結直腸癌組織中表達上調,并與腫瘤的遠處轉移及患者術后生存期短密切相關,可能作為預測結直腸癌患者預后及轉移的潛在分子標志。
[Abstract]:Colorectalcancer (CRC) is one of the most common digestive tract tumors. In recent years, the incidence of colorectal cancer in China is increasing year by year. Inflammatory bowel disease (IBD), as a recurrent and chronic inflammation, is one of the most important risk factors for colon cancer, and about 20% of the patients with colitis will eventually develop. Colitis-associated Cancer (CAC). The transformation of colitis to colon cancer is a complex process involving multiple signaling pathways and molecules. The related mechanisms are not yet clear. Serine / threonine protein kinase PLKl (polo-likekinasel) is a key regulator in the process of mammalian mitosis. The results show that PLK1 is expressed in a variety of malignant tumor tissues, including colorectal cancer (CRC), but the relationship between PLK1 and CAC has not been reported so far. Therefore, the purpose of this study is to explore the potential of PLK1 in inflammation, immune and colon cancer and to explore the role of PLK1 in CAC and to explore the role of PLK1 in CAC. In order to explore the relationship between PLK1 and CAC, this study used oxidative azo methane (azoxymethane, AOM) and sodium dextran sulfate (dextran sulfate sodium, DSS) to induce the PLK1 gene to knock into mice (PLK1fl/+ mice, experimental group) and the same nest wild type mice (WT mice, control groups). The number and load of intestinal tumor are significantly lower than that of WT control mice. It is suggested that overexpression of PLK1 can effectively inhibit the occurrence of CAC induced by AOM/DSS. Previous studies have found that PLK1 plays a role in promoting cancer in various solid tumors. Therefore, it is urgent to reveal the mechanism of PLK1 overexpression to inhibit the occurrence of CAC. We use immunohistochemistry (IHC) staining method to detect the mechanism of CAC. The expression of PLK1 in the CAC foci of the wild type mice induced by AOM/DSS was found to be highly expressed in the intestinal epithelial mesenchymal cells. Accordingly, we carried out IHC staining of PLK1 and Gr1 (labeled myeloid derived cells) in the continuous sections of the tumor tissue of colitis related colon cancer in mice, and found the positive cells of PLK1 and Gr1. There was a consistent location in the lesions of the colon. The results of immunofluorescence (IF) Co localization showed that there was a co localization phenomenon between PLK1 and Gr1 labelled medullary cells and F4/80 labeled macrophages. The results showed that PLK1 was mainly expressed in myeloid cells in the intestinal microenvironment during the occurrence of CAC. The macrophages in the microenvironment of enteritis / colon cancer Cell lines can promote the transformation of colitis to colon by secreting proinflammatory cytokines and chemokines. To further reveal the molecular mechanism of inhibiting the occurrence of CAC in the microenvironmental myeloid cells of intestinal inflammation, we use cytokine microarray to detect the colonic tissue culture of DSS treated PLK1fl/+ and WT mice. The expression of inflammatory factors in the colon was detected. The results of microchip detection showed that high expression of PLK1 could obviously inhibit the expression and secretion of multiple cytokines including TNF- alpha, including TNF- alpha and.ELISA and qRT-PCR, which confirmed the changes in the expression and secretion of TNF- alpha in the colon tissue of PLK1fl/+ mice. We further use PLK1 Specific siRNA or small molecule inhibitors treat the cultured macrophage J774.1 in vitro, and it is found that inhibition of the expression of PLK1 can obviously increase the expression of TNF- alpha. These results suggest that PLK1 may inhibit the pathogenesis and chemotaxis of myeloid cells in the intestinal inflammation microenvironment and inhibit the occurrence of CAC. The results of this study suggest that PLK1 can be used in this study. It can regulate the proinflammatory / carcinogenic function of interstitial myeloid cells, which may inhibit the occurrence of AOM/DSS induced CAC by inhibiting the expression and secretion of TNF- alpha and other inflammatory factors. This study not only helps to elucidate the new role and molecular mechanism of PLK1 in enteritis / colon related immune cells, but also may be developed for immunization in the future. The treatment strategy of environment provides theoretical basis and experimental basis. Colorectal cancer, as a common malignant tumor, is a great threat to human health and life. The recurrence and metastasis of colorectal cancer is one of the main causes of death. Therefore, it is found that the molecular markers can be used to predict the recurrence or prognosis of colorectal cancer. It is important to improve the survival rate of patients with colorectal cancer. In our laboratory, we found that KIAA1522 was abnormal expression in squamous cell carcinoma of the esophagus, lung cancer and other tumors, and was related to the prognosis of the patients. But so far, the expression changes of KIAA1522 in colorectal cancer are not clear. This study applied the tissue chip immuno group. The expression of KIAA1522 protein in 96 cases of colorectal cancer tissue and its paired cancerous normal tissues was detected by chemical staining, and the relationship between the expression changes and the clinicopathological indexes of colorectal cancer and the prognosis of the patients were statistically analyzed. The results of immunohistochemical analysis showed the main KIAA1522 protein in the colorectal epithelial cells. The positive rate of KIAA1522 protein expression was 81% (78/96) in colorectal cancer tissues and 13% (12/96) in normal tissues adjacent to the paired cancerous tissues (P0.05). Further statistical analysis showed that the overexpression of KIAA1522 protein and the three year tumor free survival of the colorectal cancer patients were three years after operation. A short and significant positive correlation (P = 0.017) and a positive correlation with distant metastasis (P = 0.012). Multiple factor Cox regression analysis showed that KIAA1522 overexpression was an independent predictor of poor prognosis in colorectal cancer patients (P= 0.020). To sum up, KIAA1522 was up-regulated in colorectal cancer tissue and distant metastasis of tumor and postoperative patients. Short term survival may be a potential molecular marker for predicting prognosis and metastasis of colorectal cancer.
【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R574.62;R735.35

【參考文獻】

相關期刊論文 前4條

1 吳李飛;郝佳潔;張彤彤;梁建偉;王征;張鈺;徐昕;蔡巖;王明榮;李敏;;UHRF1蛋白在結直腸癌中的表達情況及其預后判斷價值[J];癌變·畸變·突變;2015年01期

2 張彤彤;梁建偉;王征;張鈺;劉一臻;商利;楊海;蔡巖;徐昕;王明榮;;Cortactin蛋白在結直腸癌中的表達變化及其臨床意義[J];癌變.畸變.突變;2014年01期

3 Hugh James Freeman;;Early stage colon cancer[J];World Journal of Gastroenterology;2013年46期

4 段麗瓊;張萍;陳萬濤;;KIAA基因家族的研究進展[J];上海第二醫(yī)科大學學報;2005年04期

，

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