Long Noncoding RNAs Hepatocyte Nuclear Factor 4A Antisense RNA 1 and Hepatocyte Nuclear Factor 1A Antisense RNA 1 Are Involved in Ritonavir-Induced Cytotoxicity in Hepatoma Cells
Long Noncoding RNAs Hepatocyte Nuclear Factor 4A Antisense RNA 1 and Hepatocyte Nuclear Factor 1A Antisense RNA 1 Are Involved in Ritonavir-Induced Cytotoxicity in Hepatoma Cells
复制标题
长非编码RNA肝细胞核因子4A反义RNA 1和肝细胞核因子1A反义RNA 1参与利托那韦诱导的肝癌细胞细胞毒性
DOI：
10.1124/dmd.121.000693
复制
发表时间：
2021-12
影响因子：
3.9
通讯作者：
Lirong Zhang
中科院分区：
文献类型：
--
作者：
Xiaofei Wang;Yihang Yu;Pei Wang;Kun Yang;Yiting Wang;Liang Yan;Xiao-bo Zhong;Lirong Zhang
利托那韦（RTV）是一种用于抗HIV方案的药物增强剂，可诱导肝损伤。RTV主要通过肝脏中的细胞色素P450 3A 4（CYP 3A 4）代谢。HNF 4A反义RNA 1（HNF 4A-AS 1）和HNF 1A反义RNA 1（HNF 1A-AS 1）是调控CYP 3A 4和PXR表达的长链非编码RNA。本研究探讨了HNF 4A-AS 1和HNF 1A-AS 1在RTV诱导的肝毒性中的作用及其机制。在Huh 7和HepG 2细胞中，HNF 4A-AS 1和HNF 1A-AS 1被小发夹RNA敲低。进行乳酸脱氢酶和活性氧测定以评估RTV诱导的肝毒性。染色质免疫沉淀定量实时聚合酶链反应被用来检测PXR富集和组蛋白修饰的CYP 3A 4启动子。HNF 4A-AS 1敲低增加PXR和CYP 3A 4表达，加剧RTV诱导的细胞毒性，而HNF 1A-AS 1敲低产生相反的表型。从机制上讲，在HNF 4A-AS 1敲低后，CYP 3A 4启动子中PXR的富集和组蛋白3赖氨酸4（H3 K4 me 3）的三甲基化增加，组蛋白3赖氨酸27（H3 K27 me 3）的三甲基化减少。然而，在HNF 1A-AS 1敲低后，PXR和H3 K4 me 3富集减少。通过降低HNF 4A-AS 1或HNF 1A-AS 1引起的RTV诱导的肝毒性的改变可通过敲低或过表达PXR来逆转。HNF 4A-AS 1过表达或HNF 1A-AS 1敲低可减弱PXR激活剂利福平引起的对RTV诱导的肝损伤的易感性增加。综上所述，这些结果表明，HNF 4A-AS 1和HNF 1A-AS 1通过调节CYP 3A 4表达，主要通过影响CYP 3A 4启动子中PXR的结合和组蛋白修饰状态来调节RTV诱导的肝毒性。显著性声明HNF 4A-AS 1和HNF 1A-AS 1分别从人转录因子基因HNF 4A和HNF 1A的相邻反义基因转录，被鉴定为长非编码RNA，其可影响由利福平暴露引起的RTV诱导的肝毒性和对RTV诱导的肝毒性的易感性，主要是通过改变CYP 3A 4启动子中PXR富集和组蛋白修饰状态来影响CY 3A 4的表达。这一发现为进一步研究RTV引起肝损伤的机制提供了方向。
Ritonavir (RTV), a pharmacoenhancer used in anti-HIV regimens, can induce liver damage. RTV is primarily metabolized by cytochrome P450 3A4 (CYP3A4) in the liver. HNF4A antisense RNA 1 (HNF4A-AS1) and HNF1A antisense RNA 1 (HNF1A-AS1) are long noncoding RNAs that regulate the expression of pregnane X receptor (PXR) and CYP3A4. This study investigated the role and underlying mechanisms of HNF4A-AS1 and HNF1A-AS1 in RTV-induced hepatotoxicity. HNF4A-AS1 and HNF1A-AS1 were knocked down by small hairpin RNAs in Huh7 and HepG2 cells. Lactate dehydrogenase and reactive oxygen species assays were performed to assess RTV-induced hepatotoxicity. Chromatin immunoprecipitation quantitative real-time polymerase chain reaction was used to detect PXR enrichment and histone modifications in the CYP3A4 promoter. HNF4A-AS1 knockdown increased PXR and CYP3A4 expression and exacerbated RTV-induced cytotoxicity, whereas HNF1A-AS1 knockdown generated the opposite phenotype. Mechanistically, enrichment of PXR and trimethylation of histone 3 lysine 4 (H3K4me3) in the CYP3A4 promoter was increased, and trimethylation of histone 3 lysine 27 (H3K27me3) was decreased after HNF4A-AS1 knockdown. However, PXR and H3K4me3 enrichment decreased after HNF1A-AS1 knockdown. Alterations in RTV-induced hepatotoxicity caused by decreasing HNF4A-AS1 or HNF1A-AS1 were reversed by knockdown or overexpression of PXR. Increased susceptibility to RTV-induced liver injury caused by the PXR activator rifampicin was attenuated by HNF4A-AS1 overexpression or HNF1A-AS1 knockdown. Taken together, these results revealed that HNF4A-AS1 and HNF1A-AS1 modulated RTV-induced hepatotoxicity by regulating CYP3A4 expression, primarily by affecting the binding of PXR and histone modification status in the CYP3A4 promoter. SIGNIFICANCE STATEMENT HNF4A-AS1 and HNF1A-AS1, transcribed separately from neighboring antisense genes of the human transcription factor genes HNF4A and HNF1A, were identified as long noncoding RNAs that can affect RTV-induced hepatotoxicity and susceptibility to RTV-induced hepatotoxicity caused by rifampicin exposure, mainly by affecting the expression of CY3A4 via alterations in PXR enrichment and histone modification status in the CYP3A4 promoter. This discovery provides directions for further research on the mechanisms of RTV-induced liver injury.
登录
查看更多内容
DOI：
10.2174/187569209788654005
发表时间：
2009-06-01
影响因子：
--
作者：
Hernandez JP;Mota LC;Baldwin WS
通讯作者：
Baldwin WS
抗逆转录病毒疗法与艾滋病毒系统滥用药物之间的药物相互作用。
影响因子：
4.3
作者：
Kumar S;Rao PS;Earla R;Kumar A
通讯作者：
Kumar A
影响因子：
3.6
作者：
Liang Yan;Yiting Wang;Jingyang Liu;Ya-li Nie;Xiao-bo Zhong;Q. Kan;Lirong Zhang
通讯作者：
Liang Yan;Yiting Wang;Jingyang Liu;Ya-li Nie;Xiao-bo Zhong;Q. Kan;Lirong Zhang
影响因子：
3.9
作者：
Parimal Pande;Xiao-bo Zhong;W. Ku
通讯作者：
Parimal Pande;Xiao-bo Zhong;W. Ku
DOI：
10.32388/356hl8
发表时间：
2020-02
期刊：
Definitions
影响因子：
--
作者：
通讯作者：
--