本文選題：肺癌　切入點(diǎn)：非小細(xì)胞肺癌　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:肺癌是目前全球惡性腫瘤累及死亡的首要原因,發(fā)病率高,臨床缺乏有效治療手段,死亡率高,是嚴(yán)重危害人群健康的一大難題。惡性生長是肺癌細(xì)胞的重要生物學(xué)特征,也是評(píng)價(jià)臨床治療效果的關(guān)鍵指標(biāo),然而肺癌細(xì)胞惡性生長的維持和促進(jìn)因素仍不清楚。深入研究肺癌細(xì)胞惡性生長的維持和促進(jìn)因素,對(duì)理解肺癌發(fā)病機(jī)制和研發(fā)臨床治療新策略有重要意義。本課題以肺癌患者腫瘤細(xì)胞為靶點(diǎn),探討白細(xì)胞介素-33(IL-33)在肺癌細(xì)胞惡性生長中的作用及其可能機(jī)制。方法:腫瘤細(xì)胞分離采用腫瘤細(xì)胞分離試劑盒,細(xì)胞轉(zhuǎn)染用Lonza轉(zhuǎn)染試劑盒,轉(zhuǎn)染效率通過qPCR和流式細(xì)胞術(shù)檢測(cè)。腫瘤細(xì)胞體外生長用MTT法檢測(cè),腫瘤細(xì)胞體內(nèi)生長情況用人源化小鼠模型分析。IL-33、ST2和葡萄糖轉(zhuǎn)運(yùn)蛋白1(GLUT1)的表達(dá)水平用qPCR、免疫組化染色或流式細(xì)胞術(shù)檢測(cè),葡萄糖攝取率和乳酸產(chǎn)生水平分別按照試劑盒說明書進(jìn)行。結(jié)果:IL-33與其受體ST2在腫瘤組織中的表達(dá)水平顯著高于在癌旁組織中的表達(dá)水平;IL-33和ST2的表達(dá)水平與肺癌腫瘤分期顯著正相關(guān),在晚期肺癌患者體內(nèi)存在更高水平的表達(dá);IL-33和ST2表達(dá)水平可提示腫瘤細(xì)胞的低分化程度,在低分化肺癌細(xì)胞中存在顯著高表達(dá)。通過轉(zhuǎn)染人IL-33正義表達(dá)載體上調(diào)肺癌細(xì)胞內(nèi)IL-33表達(dá)水平可在體外和體內(nèi)有效促進(jìn)腫瘤細(xì)胞的惡性生長,通過轉(zhuǎn)染人IL-33 shRNA下調(diào)肺癌細(xì)胞內(nèi)IL-33表達(dá)水平可明顯抑制肺癌細(xì)胞在體外和體內(nèi)的惡性生長能力。IL-33蛋白刺激肺癌細(xì)胞可以直接促進(jìn)肺癌細(xì)胞在體外和體內(nèi)的惡性生長能力,上調(diào)ST2表達(dá)水平可進(jìn)一步增強(qiáng)IL-33對(duì)肺癌細(xì)胞惡性生長的促進(jìn)作用,下調(diào)ST2表達(dá)水平可阻斷IL-33對(duì)肺癌細(xì)胞惡性生長的促進(jìn)作用,ST2中和抗體亦有效抑制IL-33對(duì)肺癌細(xì)胞惡性生長的影響。IL-33影響肺癌細(xì)胞惡性生長的分子機(jī)制在于IL-33蛋白刺激肺癌細(xì)胞能夠上調(diào)腫瘤細(xì)胞膜GLUT1蛋白的水平,該過程可被ST2中和性抗體所阻斷;IL-33蛋白亦可增強(qiáng)肺癌細(xì)胞的葡萄糖攝取能力和乳酸產(chǎn)生水平;基因敲除GLUT1在肺癌細(xì)胞中的表達(dá)可顯著抑制IL-33對(duì)肺癌細(xì)胞惡性生長的促進(jìn)作用。結(jié)論:IL-33是臨床肺癌患者腫瘤進(jìn)展中的重要促癌因子,IL-33能夠維持和促進(jìn)肺癌細(xì)胞的惡性生長能力。IL-33維持和促進(jìn)肺癌細(xì)胞惡性生長可以通過ST2受體識(shí)別實(shí)現(xiàn),下調(diào)或阻斷IL-33/ST2信號(hào)可顯著抑制肺癌細(xì)胞的惡性生長。IL-33/ST2信號(hào)能夠增強(qiáng)肺癌細(xì)胞膜GLUT1蛋白水平進(jìn)而促進(jìn)肺癌細(xì)胞的葡萄糖攝取能力和糖酵解,為肺癌患者腫瘤細(xì)胞惡性生長提供能量支持�；贗L-33/ST2信號(hào)的干擾策略可能是臨床控制肺癌細(xì)胞惡性生長的有效手段。該研究不僅有助于深入理解臨床肺癌發(fā)生發(fā)展的分子機(jī)制,更可為研發(fā)臨床治療肺癌新策略提供理論依據(jù),具有重要的理論意義和現(xiàn)實(shí)意義。
[Abstract]:Objective: lung cancer is the leading cause of death from malignant tumors in the world at present. The incidence of lung cancer is high, the clinical treatment is lack of effective treatment, and the mortality rate is high. Malignant growth is an important biological feature of lung cancer cells and a key index to evaluate the clinical therapeutic effect. However, the maintenance and promotion factors of malignant growth of lung cancer cells are still unclear. It is of great significance to understand the pathogenesis of lung cancer and to develop new clinical treatment strategies. To explore the role of interleukin-33 (IL-33) in the malignant growth of lung cancer cells and its possible mechanism. Methods: tumor cells were separated by tumor cell separation kit and Lonza transfection kit was used for cell transfection. The transfection efficiency was detected by qPCR and flow cytometry. The growth of tumor cells in vitro was detected by MTT. In vivo growth of tumor cells was detected by qPCR, immunohistochemical staining or flow cytometry to analyze the expression of. IL-33 / ST2 and glucose transporter 1 (GLUT1) using a humanized mouse model. The glucose uptake rate and lactic acid production level were carried out in accordance with the kit instructions. Results the expression levels of ST2 and ST2 in tumor tissues were significantly higher than those in paracancerous tissues. The expression levels of IL-33 and ST2 in lung and lung were significantly higher than those in paracancerous tissues. There was a significant positive correlation between tumor staging and tumor staging. The higher expression of IL-33 and ST2 in patients with advanced lung cancer may indicate the low differentiation of tumor cells. The expression of IL-33 in lung cancer cells was up-regulated by transfection of human IL-33 sense expression vector, which could effectively promote the malignant growth of tumor cells in vitro and in vivo. Down-regulating the expression of IL-33 in lung cancer cells by transfection of human IL-33 shRNA could significantly inhibit the malignant growth ability of lung cancer cells in vitro and in vivo. IL-33 protein stimulated lung cancer cells can directly promote the malignant growth ability of lung cancer cells in vitro and in vivo. Upregulating the expression of ST2 can further enhance the role of IL-33 in promoting the malignant growth of lung cancer cells. Down-regulation of ST2 expression can block the effect of IL-33 on the malignant growth of lung cancer cells. ST2 neutralizing antibody can also effectively inhibit the effect of IL-33 on the malignant growth of lung cancer cells. The molecular mechanism of IL-33 affecting the malignant growth of lung cancer cells lies in the IL-33 protein prick. Lung cancer cells can up-regulate the level of GLUT1 protein in tumor cell membrane. This process can be blocked by ST2 neutralizing antibody. IL-33 protein can also enhance glucose uptake and lactic acid production in lung cancer cells. The expression of knockout GLUT1 in lung cancer cells can significantly inhibit the role of IL-33 in promoting the malignant growth of lung cancer cells. Conclusion: IL-33 is an important tumor promoting factor in clinical lung cancer patients, which can maintain and promote the growth of lung cancer cells. The ability of malignant growth. IL-33 to maintain and promote the malignant growth of lung cancer cells can be achieved by ST2 receptor recognition. Down-regulating or blocking the IL-33/ST2 signal can significantly inhibit the malignant growth of lung cancer cells. IL-33 / ST2 signal can enhance the GLUT1 protein level of lung cancer cell membrane and promote glucose uptake and glycolysis of lung cancer cells. The interference strategy based on IL-33/ST2 signal may be an effective method to control the malignant growth of lung cancer cells. This study is not only helpful to understand the molecular mechanism of the occurrence and development of clinical lung cancer. It can provide a theoretical basis for the development of new strategies for the treatment of lung cancer, and has important theoretical and practical significance.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R734.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Stephanie M Yoon;Talha Shaikh;Mark Hallman;;Therapeutic management options for stage Ⅲ non-small cell lung cancer[J];World Journal of Clinical Oncology;2017年01期

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本文編號(hào)：1647413