本文選題：人巨細胞病毒 + 轉錄激活因子5��； 參考：《青島大學》2017年碩士論文

【摘要】：神經(jīng)膠質瘤是最常見的原發(fā)性中樞神經(jīng)系統(tǒng)腫瘤,盡管采取積極的治療措施,預后依然很差。在神經(jīng)膠質瘤環(huán)境中,人巨細胞病毒(human cytomegalovirus,HCMV)基因及轉錄產(chǎn)物的表達隨著腫瘤惡性程度的增高而增高,其感染與神經(jīng)膠質瘤關系密切。轉錄激活因子5(Activating transcription factor 5,ATF5)是腫瘤標志性轉錄因子,在神經(jīng)膠質瘤中特異性高表達。本文在神經(jīng)膠質瘤環(huán)境中,探討基因沉默ATF5聯(lián)合抗病毒藥物更昔洛韋(ganciclovir,GCV)對HCMV陽性膠質瘤U87細胞增殖及病毒復制的影響。研究首先篩選合適的GCV作用濃度;利用慢病毒介導的小干擾RNA靶向干擾ATF5基因的表達,構建ATF5下調穩(wěn)定表達的LV-ATF5-RNAi U87細胞系;低感染復數(shù)(multiplicity of infection,MOI)的HCMV感染LV-ATF5-RNAi U87細胞,并用含GCV的培養(yǎng)基培養(yǎng)。在該條件下,CCK-8法檢測細胞的增殖活性;Real-time PCR、western-blot分別檢測HCMV即刻早期(IE)、早期(UL44)、晚期(UL99)基因及蛋白表達水平;空斑形成試驗檢測HCMV子代病毒復制水平。結果表明:成功構建ATF5下調穩(wěn)定表達的LV-ATF5-RNAi U87細胞�；虺聊珹TF5聯(lián)合GCV共同作用,可以較好的抑制HCMV陽性神經(jīng)膠質瘤細胞的增殖活性;基因沉默ATF5聯(lián)合GCV共同作用對HCMV在mRNA、蛋白水平、子代病毒復制水平上的抑制結果一致,都明顯大于GCV或者沉默ATF5單獨作用的抑制效率。以上結果提示,基因沉默ATF5聯(lián)合GCV可以較好的遏制HCMV病毒的復制以及神經(jīng)膠質瘤的惡性增殖作用。因此,基因沉默ATF5聯(lián)合GCV共同作用可能為臨床上治療神經(jīng)膠質瘤提供一個新的方法。
[Abstract]:Glioma is the most common primary central nervous system tumor. In glioma environment, the expression of human cytomegalovirus (HCMV) gene and transcription products of human cytomegalovirus (HCMV) increases with the increase of tumor malignancy, and its infection is closely related to glioma. Transcriptional activator 5(Activating transcription factor 5 (ATF 5) is a tumor marker transcription factor, which is specifically overexpressed in gliomas. To investigate the effects of gene silencing ATF5 combined with ganciclovirr GCVV on the proliferation and replication of HCMV positive glioma U87 cells in glioma environment. In this study, we first selected suitable GCV concentration, constructed LV-ATF5-RNAi U87 cell line, which down-regulated stable expression of LV-ATF5-RNAi U87 cell line, and HCMV infected LV-ATF5-RNAi U87 cell line with low infection complex multiplicity of infective moi, using lentivirus-mediated small interfering RNA targeted interference ATF5 gene expression, and constructed LV-ATF5-RNAi U87 cell line with down-regulated expression of ATF5-RNAi U87 cells. The culture medium containing GCV was used. Under this condition, the cell proliferation activity was detected by CCK-8 method and the expression of HCMV gene and protein were detected by real-time PCRRX western-blot, respectively, and the replication level of HCMV progeny virus was detected by plaque formation test. The results showed that ATF5 down-regulated stable expression of LV-ATF5-RNAi U87 cells. Gene silencing ATF5 combined with GCV could inhibit the proliferation of HCMV positive glioma cells. Both of them were significantly more effective than those of GCV or silencing ATF 5 alone. These results suggest that gene silencing ATF5 combined with GCV can inhibit the replication of HCMV virus and the malignant proliferation of glioma. Therefore, the combined action of gene silencing ATF 5 and GCV may provide a new method for the treatment of glioma.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R739.41

【參考文獻】

相關期刊論文 前1條

1 金銳;;巨細胞病毒特性綜述[J];科協(xié)論壇(下半月);2007年07期

，

本文編號：2029216