本文選題：人類皰疹病毒6型 + 神經(jīng)膠質(zhì)瘤��； 參考：《南京醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 人類皰疹病毒6型(Human herpesvirus 6,HHV-6)是一類嗜人淋巴細(xì)胞的雙鏈DNA病毒,于1986年由美國癌癥中心的Salahuddin等首先從淋巴增生性疾病患者外周血單個(gè)核細(xì)胞中分離到,對其進(jìn)行抗原性和酶切圖譜分析表明,它有別于其它皰疹病毒,是一種具有皰疹病毒科形態(tài)特征和嗜淋巴細(xì)胞特性的新型皰疹病毒,故命名為人類皰疹病毒6型,屬于皰疹病毒β亞科。HHV-6可分為A和B兩個(gè)亞型。HHV-6感染具有廣泛的嗜細(xì)胞性,主要侵犯CD4+T細(xì)胞、單核巨噬細(xì)胞及神經(jīng)膠質(zhì)細(xì)胞等。1988年日本學(xué)者Yamanishi確認(rèn)了HHV-6是嬰幼兒急疹(ES)的病原體,后來有報(bào)道神經(jīng)膠質(zhì)瘤、腦炎、慢性疲勞綜合癥、器官移植后感染、多發(fā)性硬化癥等多種疾病有關(guān),但對其致病機(jī)制尚不清楚。 神經(jīng)膠質(zhì)瘤(Glioma)是人類最常見的原發(fā)性腦腫瘤,到目前為止對其病因不清楚。近年來病毒在神經(jīng)膠質(zhì)瘤發(fā)生發(fā)展中的作用引起了多學(xué)科的廣泛關(guān)注。有多篇報(bào)道在神經(jīng)膠質(zhì)瘤患者瘤組織中能檢測到HHV-6 DNA的存在,但未能拿到病毒。人神經(jīng)膠質(zhì)瘤U251細(xì)胞是一種星形膠質(zhì)瘤細(xì)胞株,具有星形膠質(zhì)細(xì)胞部分特性,因此若能夠建立HHV-6體外感染U251細(xì)胞模型,對研究HHV-6與神經(jīng)膠質(zhì)瘤的關(guān)系有指導(dǎo)意義。 本論文分三個(gè)部分,(一)HHV-6GS株在CBMCs中的培養(yǎng)擴(kuò)增及病毒效價(jià)的滴定。通過PCR檢測到HHV-6的DNA,間接免疫熒光檢測到HHV-6抗原。病毒的滴度最終確定約為5×103TCID50/ml。(二)HHV-6感染神經(jīng)膠質(zhì)瘤U251細(xì)胞建立體外感染模型。結(jié)果發(fā)現(xiàn)HHV-6能感染U251細(xì)胞引起細(xì)胞病變,促進(jìn)細(xì)胞增殖,使細(xì)胞周期發(fā)生改變及細(xì)胞因子IL-6分泌增加;成功建立了體外感染的模型。(三)對有絲分裂原活化蛋白激酶(mitogen-activated protein kinase , MAPK)信號(hào)通路的分支JNK和P38通路進(jìn)行研究。結(jié)果發(fā)現(xiàn)在HHV-6的作用下磷酸化JNK和P38蛋白表達(dá)增加,病毒感染可能通過活化JNK和P38信號(hào)通路,引起腫瘤細(xì)胞的分化。 在成功建立HHV-6感染U251細(xì)胞體外模型的基礎(chǔ)上,對HHV-6感染神經(jīng)膠質(zhì)瘤U251細(xì)胞的功能及機(jī)制作了初步研究,結(jié)果說明HHV-6與神經(jīng)膠質(zhì)瘤的發(fā)生發(fā)展存在一定的聯(lián)系,為進(jìn)一步研究神經(jīng)膠質(zhì)瘤的病毒病因和發(fā)病機(jī)理提供理論和實(shí)驗(yàn)依據(jù)。
[Abstract]:Human herpesvirus 6 (HHV-6) is a human lymphocytic double-stranded DNA virus. It was first isolated from peripheral blood mononuclear cells (PBMC) of patients with lymphoproliferative diseases by Salahuddin et al of the American Cancer Center in 1986. The results of antigenicity and restriction endonuclease analysis showed that it was a new type of herpesvirus, which was different from other herpesviruses and had morphologic and lymphophilic characteristics of herpesviridae, so it was named human herpesvirus type 6. The infection of herpesvirus 尾 subfamily. HHV-6 can be divided into A and B subtypes. HHV-6 infection has a wide range of cytophilia, mainly invading CD4 T cells. In 1988, the Japanese scholar Yamanishi confirmed that HHV-6 was the pathogen of infantile rash ES.After that, glioma, encephalitis, chronic fatigue syndrome, infection after organ transplantation were reported. Multiple sclerosis and other diseases are related, but its pathogenesis is not clear. Glioma (Glioma) is the most common primary brain tumor in humans. In recent years, the role of virus in the development of glioma has attracted wide attention. The presence of HHV-6 DNA was detected in glioma tissues, but the virus was not obtained. Human glioma U251 cell line is a kind of astroglioma cell line, which has some astrocytic properties. Therefore, if HHV-6 can be established to infect U251 cells in vitro, it will be helpful to study the relationship between HHV-6 and glioma. This paper is divided into three parts: culture and amplification of HHV-6GS strain in CBMCs and titration of virus titer. HHV-6 was detected by PCR and HHV-6 antigen was detected by indirect immunofluorescence. The titer of the virus was determined to be about 5 脳 103 TCID 50 / ml. (HHV-6 infected glioma U251 cells) in vitro infection model was established. The results showed that HHV-6 could induce cytopathic changes, promote cell proliferation, change cell cycle and increase the secretion of cytokine IL-6 in U251 cells, and the model of infection in vitro was successfully established. (3) the branching JNK and P38 pathways of mitogen-activated protein kinase, MAPK) signaling pathway were studied. The results showed that the expression of phosphorylated JNK and P38 protein increased under the action of HHV-6. Virus infection may induce the differentiation of tumor cells by activating JNK and P38 signaling pathway. On the basis of successfully establishing HHV-6 infected U251 cell model in vitro, the function and mechanism of HHV-6 infected glioma U251 cells were studied. The results showed that there was a certain relationship between HHV-6 and the occurrence and development of glioma. To provide theoretical and experimental basis for further study of viral etiology and pathogenesis of glioma.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R392

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