乙丙肝病毒相互干扰及适配体在肝癌诊断及靶向治疗中的作用

发布时间：2018-05-09 13:12

本文选题：乙型肝炎病毒(HBV) + 丙型肝炎病毒(HCV)　；参考：《吉林大学》2016年博士论文

【摘要】：第一部分:乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)感染是导致全球肝脏疾病的主要原因。这项研究的目的是评估HBV-HCV合并感染的临床及病毒学特征。本研究共收集3238名中国丙型肝炎病毒高发地区的当地居民的人口数据。实验室检查包括anti-HCV、乙肝病毒血清学指标,肝功能检查,以及血常规分析等血清学指标。我们还对anti-HCV阳性的患者检测HCV RNA及HCV基因分型,对HBs Ag阳性的患者检测HBV DNA。第一部分实验结果如下:被调查人群中共有1468人感染乙型肝炎病毒或丙型肝炎病毒。其中单纯HCV感染组1200人,单纯HBV感染组161人,HBV-HCV合并感染组107人。HBV-HCV合并感染组HBV DNA阳性率(84.1%)低于单纯HBV感染组(94.4%,P0.001)。HBV-HCV合并感染组HBV DNA水平与单纯HBV感染组相比显著降低(1.97[IQR,1.3-3.43]versus 3.06[IQR,2-4.28]Log10 IU/m L;P0.001)。此外,HBV-HCV合并感染组HCV RNA阳性率(53.3%)低于单纯HCV感染组(86.9%,P0.001),其HCV RNA水平与单纯HCV感染组相比显著降低(1.18[IQR,0-5.57]versus 5.87[IQR,3.54-6.71]Log10 IU/ml;P0.001)。HBV-HCV合并感染组ALT,AST,ALP,GGT,APRI和FIB-4水平高于单纯HBV感染组,但ALB,血小板水平低,但HBV-HCV合并感染组结果与单纯HCV感染组相似。总之,这些结果表明,在HBV-HCV合并感染组中HCV和HBV存在相互抑制病毒复制。根据HBV-HCV合并感染组患者的血清学显示,其与单纯HCV感染组相比肝损伤程度相似,但比单纯HBV感染组肝损伤程度更重。第二部分:肝细胞癌(HCC)是全世界肿瘤死亡第三大原因,每年有50万人死于肝细胞癌(HCC)。虽然化学疗法已被广泛用于治疗HCC患者,但化疗药物副作用严重,选择性递送治疗抗癌药物的疗法正在被广泛研究。在这方面,基于适配体的肿瘤靶向药物递送已成为一个有前途的方法,以增加化疗药物的疗效,并减少或消除药物毒性。本研究我们通过指数富集配体系统进化(SELEX)的方法开发了一种新的特异性识别Hep G2细胞的适配体(HCA#3),并利用HCA#3作为靶向配体,将阿霉素选择性递送到体外培养的Hep G2细胞中。第二部分实验结果如下:筛选的76个碱基的核苷酸适配体特异结合Hep G2肝癌细胞,但不与对照细胞结合。我们用重复的CG结构对适配体HCA#3做了修饰,使其具备携带多个阿霉素的能力。每个修饰过的HCA#3适配体可以插入四个阿霉素分子,从而形成适配体-阿霉素偶合物(Ap DC)。生物稳定性分析表明,Ap DC分子在血清中稳定。功能分析表明,Ap DC特异性靶向并将阿霉素释放到Hep G2细胞中。HCA#3 APDC诱导Hep G2细胞凋亡,但对对照细胞影响很小。总之,这些结果表明,HCA#3 Ap DC是一种很有前途的靶向治疗的适配体,可以将阿霉素特异高效的释放到肝癌细胞中。
[Abstract]:Part I: hepatitis B virus (HBV) and hepatitis C virus (HCV) infection are the main causes of global liver disease. The aim of this study was to assess the clinical and virological characteristics of HBV-HCV coinfection. In this study, data were collected from 3238 local residents in high incidence areas of hepatitis C virus in China. Laboratory tests include anti-HCV, hepatitis B serological markers, liver function tests, and routine serological analysis. We also detected HCV RNA and HCV genotyping in anti-HCV positive patients and HBV DNA in HBs Ag positive patients. The results of the first part are as follows: 1468 people were infected with hepatitis B virus or hepatitis C virus. The positive rate of HBV DNA in 1200 patients with HCV infection alone and 107 patients with HBV-HCV co-infection with HBV-HCV infection group was 84.1). The level of HBV DNA in HBV-HCV co-infection group was significantly lower than that in HBV infection group. The HBV DNA level of HBV-HCV co-infection group was 1.97 [IQR1.3-3.43] versus 3.06 [IQRN2-4.28] Log10 IU/m LGP 0.001P ~ (0.001). In addition, the positive rate of HCV RNA in HBV-HCV co-infection group (53.3) was lower than that in simple HCV infection group (P 0.001). The HCV RNA level in HBV-HCV co-infection group was significantly lower than that in HCV infection group, but the level of versus 5.87 [IQRN 3.54-6.71] Log10 IUP / ml P0.001. HBV-HCV co-infection group was significantly lower than that in HBV infection group. However, the results of HBV-HCV combined with infection group were similar to those of simple HCV infection group. All in all, these results suggest that HCV and HBV inhibit viral replication in the HBV-HCV co-infection group. According to the serology of patients with HBV-HCV combined with infection, the degree of liver injury was similar to that of HCV infection group, but the degree of liver injury was more serious than that of simple HBV infection group. The second part: hepatocellular carcinoma (HCC) is the third leading cause of cancer death in the world. 500000 people die from HCC every year. Although chemotherapy has been widely used in the treatment of HCC patients, the side effects of chemotherapeutic drugs are serious, and selective delivery therapy for anticancer drugs is being widely studied. In this regard, aptamer based tumor targeting drug delivery has become a promising method to increase the efficacy of chemotherapeutic drugs and reduce or eliminate drug toxicity. In this study, we developed a novel aptamer for Hep G2 cells by exponential enrichment ligand phylogeny (SELEX), and used HCA#3 as a target ligand to selectively deliver adriamycin to Hep G2 cells cultured in vitro. The results of the second part were as follows: the 76 base nucleotide aptamers specifically bound to Hep G2 hepatoma cells, but did not bind to the control cells. The aptamer HCA#3 was modified with repeated CG structure to carry more than one adriamycin. Each modified HCA#3 aptamer can be inserted into four adriamycin molecules to form an aptamer-adriamycin coupling compound. The stability analysis showed that AP-DC was stable in serum. Functional analysis showed that AP-DC specifically targeted and released adriamycin into Hep G2 cells. HCA#3 APDC induced apoptosis of Hep G2 cells, but had little effect on the control cells. In conclusion, these results suggest that HCA#3Ap DC is a promising aptamer for targeted therapy and can release adriamycin specifically and efficiently into hepatoma cells.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.7

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