發(fā)布時間：2018-03-29 14:44

  本文選題：食管鱗癌　切入點：PKM2　出處：《山西醫(yī)科大學(xué)》2017年博士論文

【摘要】：食管鱗狀細(xì)胞癌(ESCC)是一種地區(qū)發(fā)病率高、治愈率低、生存率低,且預(yù)后差的惡性腫瘤。其代謝、增殖與轉(zhuǎn)移涉及眾多信號通路。這些信號通路相互交叉連接,形成復(fù)雜的信號通路網(wǎng),使得食管鱗癌的治療效果并不如預(yù)期。文獻(xiàn)表明,通過調(diào)節(jié)食管鱗癌組織的能量代謝來控制腫瘤細(xì)胞的生長,可以作為一種新的腫瘤治療方法進(jìn)行探索和研究,對開發(fā)新的治療策略具有非常重要的意義。已有研究表明,不同于正常組織細(xì)胞,腫瘤組織細(xì)胞無論氧氣充足還是缺氧,都采用糖酵解的方式獲取能量,即有氧糖酵解。M2型丙酮酸激酶(PKM2)是腫瘤細(xì)胞有氧糖酵解過程中最后一步的關(guān)鍵酶,是它將磷酸烯醇式丙酮酸(PEP)分解為丙酮酸,從而使各種癌細(xì)胞獲取能量。研究表明,PKM2不僅在腫瘤細(xì)胞代謝中發(fā)揮重要作用,而且在腫瘤細(xì)胞的增殖、轉(zhuǎn)化以及預(yù)后中也是重要的信號分子,但是PKM2在腫瘤細(xì)胞中被調(diào)節(jié)的機制有待進(jìn)一步研究。m TOR信號通路調(diào)節(jié)細(xì)胞功能,影響蛋白質(zhì)的合成,在多種惡性腫瘤中發(fā)現(xiàn)其高度活躍。其下游信號分子可調(diào)節(jié)多種轉(zhuǎn)錄因子影響腫瘤細(xì)胞中關(guān)鍵酶的形成。在肝癌和腎臟腫瘤中發(fā)現(xiàn),m TOR可以調(diào)節(jié)PKM2的表達(dá)。所以,設(shè)計本課題,觀察在食管鱗癌中m TOR信號通路是否可以調(diào)節(jié)PKM2的表達(dá),以及是否可以以此影響腫瘤細(xì)胞的能量代謝。首先通過病理切片的免疫組化染色確定在食管鱗癌中m TOR和PKM2的表達(dá)均上調(diào)。通過改變m TOR和PKM2的表達(dá)量來研究其對食管鱗癌細(xì)胞KYSE150能量代謝的影響,證明m TOR可以調(diào)節(jié)PKM2進(jìn)而影響食管鱗癌細(xì)胞的有氧糖酵解,為開發(fā)食管鱗癌新的治療策略,提供依據(jù)和思路。目的:1、研究PKM2對食管鱗癌細(xì)胞有氧糖酵解的影響。2、研究m TOR信號通路對食管鱗癌細(xì)胞有氧糖酵解的影響。3、研究在食管鱗癌細(xì)胞中m TOR通路調(diào)節(jié)PKM2影響有氧糖酵解代謝的機制。方法:第一部分:食管鱗癌患者組織樣本及數(shù)據(jù)分析為驗證食管鱗癌患者組織中PKM2及p-m TOR的表達(dá)情況,選取食管鱗癌及非腫瘤食管組織樣本各30例,并充分收集患者的臨床病理組織及資料進(jìn)行免疫組化實驗,檢測PKM2及p-m TOR在食管鱗癌及非腫瘤食管組織中的表達(dá)情況。第二部分:PKM2的表達(dá)影響食管鱗癌細(xì)胞的有氧糖酵解1、檢測PKM2下調(diào)對食管鱗癌細(xì)胞KYSE150葡萄糖消耗和乳酸生成的影響,并與非腫瘤食管組織進(jìn)行對比。2、過表達(dá)食管鱗癌細(xì)胞系KYSE150細(xì)胞中的PKM2,檢測PKM2過表達(dá)對食管鱗癌細(xì)胞葡萄糖消耗和乳酸生成情況比較的影響。第三部分:抑制或者活化m TOR信號通路對食管鱗癌細(xì)胞有氧糖酵解的影響1、m TOR抑制劑雷帕霉素作用于KYSE150細(xì)胞,觀察其對m TOR信號通路以及對細(xì)胞有氧糖酵解的影響。2、下調(diào)m TOR抑制基因PTEN對m TOR信號通路以及細(xì)胞有氧糖酵解的影響。第四部分:在食管鱗癌KYSE150細(xì)胞系中,m TOR通路通過對PKM2的調(diào)節(jié)影響KYSE150細(xì)胞的有氧糖酵解1、m TOR抑制劑雷帕霉素作用于KYSE150細(xì)胞對PKM2表達(dá)量的影響。2、抑制基因PTEN抑制組對PKM2表達(dá)以及細(xì)胞糖酵解作用的影響。3、PKM2被敲低后,抑制PTEN基因(即活化m TOR通路)也不能上調(diào)食管鱗癌細(xì)胞的有氧糖酵解作用。結(jié)果:1、m TOR和PKM2在食管鱗癌組織中表達(dá)升高。2、PKM2在食管鱗癌細(xì)胞代謝中對有氧糖酵解有調(diào)節(jié)作用。3、m TOR在食管鱗癌細(xì)胞代謝中對有氧糖酵解有調(diào)節(jié)作用。4、在食管鱗癌細(xì)胞中m TOR通路通過調(diào)節(jié)PKM2影響有氧糖酵解。結(jié)論:本實驗初步闡明PKM2和m TOR在食管鱗癌組織中均呈現(xiàn)高表達(dá)狀態(tài),m TOR在食管鱗癌細(xì)胞中可以調(diào)節(jié)PKM2,繼而影響食管鱗癌細(xì)胞的有氧糖酵解,為食管鱗癌的治療提供了新的策略。
[Abstract]:Esophageal squamous cell carcinoma (ESCC) is an area of high incidence, low cure rate, low survival rate and poor prognosis of malignant tumor. Its metabolism, proliferation and metastasis involves many signaling pathways. These pathways cross connect with each other, forming a complex network of signaling pathways, the therapeutic effect of esophageal squamous cell carcinoma is not as expected. Literature shows that, through the regulation of energy metabolism in human esophageal squamous cell carcinoma to control the growth of tumor cells, can be used as a new tumor treatment methods of exploration and research, has very important significance for the development of new therapeutic strategies. Studies have shown that different from normal cells, tumor cells regardless of sufficient oxygen or oxygen and use glycolysis way to obtain energy, namely the aerobic glycolysis of.M2 pyruvate kinase (PKM2) is a key enzyme in tumor cells of oxygen during glycolysis is the last step, it will be allyl phosphate Alcohol type pyruvate (PEP) decomposition of pyruvate, so that a variety of cancer cells to obtain energy. The results show that PKM2 not only play an important role in the metabolism of tumor cells, and the proliferation of tumor cells, but also an important signal molecule transformation and prognosis, but the mechanism of PKM2 is regulated in tumor cells for further study of.M TOR the signaling pathways that regulate cell function, affect protein synthesis, the highly active in a variety of malignant tumors. The downstream signaling molecules can regulate many transcription factors influence the formation of key enzymes in tumor cells. Found in liver cancer and kidney cancer, the expression of M TOR PKM2 can be adjusted. Therefore, the subject of the design, to observe the expression in esophageal squamous cell carcinoma m TOR signal pathway can regulate PKM2, and whether this can affect energy metabolism of tumor cells. The immunohistochemical staining of pathological sections To determine the expression of M TOR and PKM2 were up-regulated in esophageal squamous cell carcinoma. The expression change of M TOR and PKM2 to study its effect on esophageal squamous cell carcinoma KYSE150 cell energy metabolism, TOR proved that M can regulate PKM2 and esophageal squamous cell carcinoma of aerobic glycolysis, as a new strategy of development of esophageal squamous cell carcinoma treated with the basis and ideas. Objective: 1. Study on PKM2 effect of aerobic glycolysis of.2 for esophageal squamous cell carcinoma, m TOR signal pathway effects of aerobic glycolysis of.3 for esophageal squamous cell carcinoma, research in esophageal squamous cell carcinoma m TOR pathway in regulation of PKM2 can influence the mechanism of oxygen glycolytic metabolism. Methods: the first part: the sample and data analysis for patients with esophageal squamous cell carcinoma tissue expression validation of esophageal squamous cell carcinoma PKM2 and P-M TOR, selection of esophageal squamous cell carcinoma and non esophageal tumor tissue samples of the 30 cases, and the full collection of clinical and pathological tissues of patients Data and immunohistochemical experiments, to detect the expression of PKM2 and P-M TOR in esophageal squamous cell carcinoma and non tumor tissues. The second part: the effect of PKM2 expression in esophageal squamous cell carcinoma of aerobic glycolysis 1, detection of PKM2 downward effect on esophageal squamous cell carcinoma cell line KYSE150 glucose consumption and lactic acid, and compared.2 and esophageal tissue non tumor, overexpression of KYSE150 cells in esophageal squamous cell carcinoma PKM2 cell line, detect the effects of PKM2 overexpression on esophageal squamous cell carcinoma cell line comparison of glucose consumption and lactic acid production situation. The third part: the inhibition or activation of M TOR signaling pathway in esophageal squamous cell carcinoma cells effect of aerobic glycolysis inhibitor 1, m TOR the effect of rapamycin on KYSE150 cells and observe the M and TOR signal pathway on cell effect of aerobic glycolysis of.2, down-regulation of M TOR gene PTEN aerobic glycolysis effect on M cells and TOR signal pathway Ring. The fourth part: in esophageal squamous cell carcinoma cell line KYSE150, to regulate the PKM2 effect of KYSE150 cells in aerobic glycolysis 1 through the m TOR pathway, m TOR inhibitor rapamycin in KYSE150 cells the expression of.2 on the PKM2 effect of.3 suppressor gene PTEN inhibited group solution effect on PKM2 expression and cell sugar fermentation, knockdown of PKM2 inhibited PTEN gene (i.e., activation of M TOR pathway in esophageal squamous cell carcinoma) also can not increase the aerobic glycolysis. Results: 1 the expression of M, TOR and PKM2 in esophageal squamous cell carcinoma increased.2, PKM2 in esophageal squamous cell carcinoma on the metabolism of aerobic glycolysis is the role of.3 in regulation of M, TOR in esophageal squamous cell carcinoma on the metabolism of aerobic glycolysis of the regulatory role of.4 in esophageal squamous cell carcinoma m TOR pathway by regulating the PKM2 effect of aerobic glycolysis. Conclusion: the PKM2 and m TOR were preliminarily showed high expression in esophageal squamous cell carcinoma. State of state, m TOR can regulate PKM2 in esophageal squamous cell carcinoma cells, and then influence the aerobic glycolysis of esophageal squamous cell carcinoma cells, which provides a new strategy for the treatment of esophageal squamous cell carcinoma.

【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R735.1

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