THE EFFECT OF DNA DEMETHYLATION HIV-1 EXPRESSION IN VITRO

DNA 去甲基化对 HIV-1 体外表达的影响

• 批准号: 3853267
• 负责人: H MITSUYA
• 金额: --
• 依托单位: DIVISION OF CANCER TREATMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3853267
• 关键词: DNA methylation; HIV infections; T lymphocyte; azacitidine; gene expression; human immunodeficiency virus 1; provirus; southern blotting; tissue /cell culture

The effect of DNA demethylation on human immunodeficiency virus type-1 (H-1)IV expression was investigated in several cell lines infected with HIV-1 in vitro. In order to demethylate proviral DNA, 5-azacytidine (AZC) and its analog were employed. A persistently HIV-1-infected T-cell line, ACH2, produced a low level of HIV-1 in vitro. However, following exposure to AZC or its analog, a profound potentiation of viral expression occurred as assessed by syncytia formation and production of reverse transcriptase and p24 Gag protein. Alternation in the methylation pattern of proviral DNA in ACH2 was confirmed by Southern blot hybridization using methylation sensitive/insensitive isoschizomer restriction enzymes. AZC-induced potentiation of viral expression did not occur in any of high HIV-1-producers, chronically HIV-1 infected H9 cells, subacutely HIV-1-infected normal CD4+ T-cells, or subacutely infected lectin-activated peripheral blood mononuclear cells. Southern blot hybridization revealed that proviral DNA in these highly HIV-1-producing cells had already been extensively demethylated, and no altered DNA methylation pattern had occurred following AZC treatment. We also found that the inherent toxicity of AZC per se was, at least in part, associated with the observed enhancement of HIV-1 expression in ACH2 cells. The present data suggest that DNA methylation may play a role in regulating HIV-1 expression at least under certain circumstances. Further study is also warranted to test the possibility that certain therapeutics which exert toxicity to cells such as anticancer agents may affect the expression of HIV-1 in patients with HIV-1 infection and neoplasms.

DNA去甲基化对人类免疫缺陷病毒1型的影响 在感染了以下病毒的几种细胞系中研究了（H-1）IV表达： 体外HIV-1。为了使前病毒DNA去甲基化， (AZC)及其类似物。 持续感染HIV-1的 T细胞系ACH 2在体外产生低水平的HIV-1。 然而，在这方面， 暴露于AZC或其类似物后， 通过合胞体形成评估病毒表达， 逆转录酶和p24 Gag蛋白的产生。 交替 ACH 2中前病毒DNA的甲基化模式通过 使用甲基化敏感/不敏感的Southern印迹杂交 异构体限制酶。 AZC诱导的病毒表达增强在任何 高HIV-1生产者，慢性HIV-1感染的H9细胞，亚急性 HIV-1感染的正常CD 4 + T细胞，或亚急性感染 凝集素激活的外周血单核细胞。 Southern印迹 杂交显示，在这些HIV-1高产细胞中， 细胞已经被广泛地去甲基化， AZC处理后出现甲基化模式。 我们还发现 AZC本身的固有毒性，至少部分， 与观察到的ACH 2中HIV-1表达增强相关 细胞 目前的数据表明，DNA甲基化可能在 至少在某些情况下调节HIV-1表达。 进一步的研究也是必要的，以测试某些 对细胞产生毒性的治疗剂如抗癌剂可 影响HIV-1感染患者中HIV-1的表达， 肿瘤。