本文關(guān)鍵詞： EBV 病毒裂解感染誘導療法 蟲草素 阿霉素 C/EBPβ　出處：《中國農(nóng)業(yè)大學》2017年博士論文　論文類型：學位論文

【摘要】：EB病毒(Epstein-Barrvirus,EBV)是1964發(fā)現(xiàn)的一種人類γ皰疹病毒,EBV在全球人群中的感染率高達95%以上,并以潛伏感染狀態(tài)在宿主體內(nèi)終身攜帶。EBV感染與多種人類疾病相關(guān),而且EBV是第一種被發(fā)現(xiàn)的腫瘤相關(guān)病毒。全球每年20萬例新發(fā)腫瘤與EBV相關(guān),并且1.8%左右的癌癥死亡是由EBV相關(guān)腫瘤所導致。免疫抑制和免疫缺陷人群更易發(fā)生EBV陽性腫瘤;與發(fā)達地區(qū)相比,發(fā)展中地區(qū)具有更高的EBV相關(guān)腫瘤發(fā)生率。我國是EBV陽性腫瘤高發(fā)國度,尤其是鼻咽癌和胃癌,這對人民健康造成巨大威脅。鑒于EBV陽性腫瘤細胞內(nèi)多含有處于潛伏感染狀態(tài)的EBV,而且EBV同時是一種溶瘤病毒,這便提供了通過誘導EBV進入裂解感染狀態(tài)而殺傷腫瘤細胞的治療方式。該治療方式已被多項臨床實驗證實行之有效,可以明顯改善患者的生存質(zhì)量并延長生存時間。然而目前臨床上可供選用的誘導EBV裂解感染的藥物還十分有限,因此亟需開發(fā)新的藥物用于治療EBV陽性腫瘤。蟲草素(cordycepin)是從傳統(tǒng)中藥冬蟲夏草中提取的單體小分子化合物,具有抗炎、抗腫瘤和免疫調(diào)節(jié)等多種生物學作用。本文通過研究發(fā)現(xiàn),蟲草素可以通過PKC、p38 MAPK以及MEK等信號通路激活腫瘤細胞內(nèi)的EBV進入裂解感染期,并且蟲草素對EBV陽性腫瘤細胞具有一定的選擇殺傷作用。聯(lián)合作用實驗顯示,蟲草素可以明顯增強傳統(tǒng)化療藥物阿霉素(doxorubicin)激活EBV裂解感染的能力,同時蟲草素可以顯著增強阿霉素對EBV陽性腫瘤細胞的殺傷能力。通過分子機制研究發(fā)現(xiàn),蟲草素和阿霉素聯(lián)合使用可以激活細胞內(nèi)的PKC-p38 MAPK信號通路,并介導EBV轉(zhuǎn)錄激活因子BZLF-1的轉(zhuǎn)錄表達。進一步研究發(fā)現(xiàn)蟲草素和阿霉素在磷酸化激活p38 MAPK以及下游轉(zhuǎn)錄因子C/EBPβ的過程中具有聯(lián)合增強作用。結(jié)合以往實驗和序列分析,發(fā)現(xiàn)在BZLF-1的啟動子Zp上具有多個C/EBPβ結(jié)合位點;通過熒光素酶報告載體實驗和siRNA敲低實驗研究發(fā)現(xiàn),轉(zhuǎn)錄因子C/EBPβ介導BZLF-1轉(zhuǎn)錄以及EBV裂解感染激活過程。最后,動物實驗表明蟲草素和阿霉素聯(lián)合使用對EBV陽性腫瘤具有明顯的治療作用,不僅可以明顯抑制小鼠體內(nèi)EBV陽性腫瘤的形成和生長,而且蟲草素可以明顯減緩阿霉素所造成的小鼠體重下降問題。以上研究結(jié)果表明,天然小分子化合物蟲草素可以誘導EBV裂解感染,而且蟲草素和傳統(tǒng)化療藥物阿霉素聯(lián)合使用對于激活EBV裂解感染具有明顯的聯(lián)合增強作用,體內(nèi)實驗證實蟲草素和阿霉素聯(lián)合使用對于EBV陽性腫瘤具有明顯的治療效果,本文同時闡明了蟲草素與阿霉素聯(lián)合使用通過PKC-p38 MAPK-C/EBPβ信號通路激活EBV裂解感染的分子機制。以上研究結(jié)果為EBV陽性腫瘤的治療提供了重要實驗依據(jù)和臨床應用參考。
[Abstract]:Epstein-Barr virus (EBV) is a human gamma-herpesvirus (EBV) found in 1964. The infection rate of EBV in the global population is as high as 95%. Epstein-Barrvirus.EBV infection in the host is associated with many human diseases. EBV is the first tumor-associated virus found in the world. 200,000 new tumors are associated with EBV every year, and about 1.8% of cancer deaths are caused by EBV related tumors. Immunosuppressive and immunodeficient people are more likely to develop EBV positive tumors. Compared with the developed regions, the developing regions have a higher incidence of EBV related tumors. China is a country with high incidence of EBV positive tumors, especially nasopharyngeal carcinoma and gastric cancer. This poses a great threat to people's health. Given that EBV positive tumor cells often contain potentially infected EBVs, and EBV is also a tumor lysate virus, This provides a way to kill tumor cells by inducing EBV to enter the state of lytic infection, which has been proven to be effective in many clinical trials. It can significantly improve the quality of life and prolong the life span of patients. However, there are very few drugs available in clinic to induce EBV lytic infection. Therefore, it is urgent to develop new drugs for the treatment of EBV positive tumors. Cordycepin (Cordycepin) is a small monomer compound extracted from traditional Chinese medicine Cordycepin, which has anti-inflammatory properties. In this paper, we found that Cordycepin can activate the EBV in tumor cells to enter the lytic infection stage through signal pathways such as PKCnp38 MAPK and MEK. In addition, Cordycepin has a selective killing effect on EBV positive tumor cells. The combined effect of Cordycepin on EBV positive tumor cells showed that Cordycepin could significantly enhance the ability of traditional chemotherapeutic drug doxorubicin to activate EBV lytic infection. At the same time, Cordycepin could significantly enhance the cytotoxicity of adriamycin to EBV positive tumor cells. It was found that the combination of Cordycepin and Adriamycin could activate the PKC-p38 MAPK signaling pathway in the cells. Furthermore, it was found that Cordycepin and adriamycin could enhance the phosphorylation of p38 MAPK and downstream transcription factor C / EBP 尾, combining with previous experiments and sequence analysis. It was found that there were several C / EBP 尾 binding sites on the promoter Zp of BZLF-1. Through luciferase report vector experiment and siRNA knockdown experiment, it was found that the transcription factor C / EBP 尾 mediated BZLF-1 transcription and the activation of EBV lytic infection. Animal experiments showed that the combination of cordycepin and adriamycin had a significant therapeutic effect on EBV positive tumors, and could not only inhibit the formation and growth of EBV positive tumors in mice. In addition, Cordycepin can significantly reduce the weight loss caused by adriamycin in mice. The above results show that Cordycepin, a natural small molecular compound, can induce EBV cleavage infection. In addition, the combination of Cordycepin and adriamycin in combination with traditional chemotherapeutic drugs can significantly enhance the activation of EBV lytic infection. In vivo, the combination of Cordycepin and Adriamycin has a significant therapeutic effect on EBV positive tumors. The molecular mechanism of the combination of cordycepin and adriamycin to activate EBV lytic infection through PKC-p38 MAPK-C/EBP 尾 signaling pathway was also elucidated. These results provide an important experimental basis and clinical application reference for the treatment of EBV positive tumors.
【學位授予單位】：中國農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R730.5

【參考文獻】

相關(guān)期刊論文 前4條

1 張超;鐘沁;張錫峰;胡代曦;何雪梅;李權(quán)倫;馮濤;;蟲草素對A549細胞增殖、凋亡及核轉(zhuǎn)錄因子NF-κB活性的影響[J];中藥材;2015年04期

2 Lili Li;Yuan Zhang;Bing-Bing Guo;Francis K.L.Chan;Qian Tao;;Oncogenic induction of cellular high CpG methylation by Epstein-Barr Barr virus in malignant epithelial cells[J];Chinese Journal of Cancer;2014年12期

3 CHEN Ying-hui;HAO Lyh-Jyh;HUNG Chih-peng;CHEN Jung-wei;LEU Sew-fen;HUANG Bu-miin;;Apoptotic Effect of Cisplatin and Cordycepin on OC3 Human Oral Cancer Cells[J];Chinese Journal of Integrative Medicine;2014年08期

4 Massimo De Paschale;Pierangelo Clerici;;Serological diagnosis of Epstein-Barr virus infection: Problems and solutions[J];World Journal of Virology;2012年01期

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本文編號：1520456