lncRNA PVT1 promotes hepatitis B virus-positive liver cancer progression by disturbing histone methylation on the c-Myc promoter
lncRNA PVT1 promotes hepatitis B virus-positive liver cancer progression by disturbing histone methylation on the c-Myc promoter
复制标题
DOI：
10.3892/or.2019.7444
复制
发表时间：
2020-02-01
期刊：
影响因子：
4.2
通讯作者：
Lu, Wei
中科院分区：
文献类型：
--
作者：
Jiang, Bei;Yang, Bing;Lu, Wei
长链非编码RNA（LncRNA）PVT 1最近被报道参与肝细胞癌（HCC）的发展。我们的目的是阐明PVT 1与临床上B型肝炎病毒阳性肝癌的相关性，以及PVT 1在肝癌细胞生物学中的作用，并研究其潜在的分子机制。进行qPCR分析以检查PVT 1在B型肝炎病毒阳性HCC组织和肝癌细胞系中的表达。通过转染过表达载体或shRNA实现lncRNA PVT 1过表达和敲低。相应地检查细胞增殖、集落形成、迁移、凋亡和侵袭能力。RNA pull-down检测PVT 1与PRC 2复合物之间的连接。采用染色质免疫沉淀法检测MYC启动子上EZH 2蛋白和H3 K27 me 3的结合水平。结果显示，与HBV阴性样本相比，在B病毒阳性的HCC组织中检测到PVT 1的上调。在B型肝炎病毒阳性的Hep 3 B细胞中，而不是在B型肝炎病毒阴性的HepG 2细胞中，lncRNA PVT 1增强细胞增殖、迁移和侵袭。PVT 1能够与EZH 2结合，阻断EZH 2募集至MYC启动子，从而通过改变H3 K37 me 3的状态促进肝癌细胞MYC的表达，EZH 2蛋白与lncRNA-PVT 1表达呈负相关。总之，我们的研究结果表明，lncRNA PVT 1通过干扰MYC启动子上的组蛋白甲基化促进B型肝炎病毒阳性肝癌的进展，这表明lncRNA PVT 1可能是开发早期B型肝炎病毒阳性肝癌诊断和治疗策略的潜在靶点。
Long noncoding RNA (LncRNA) PVT1 has recently been reported to be involved in the development of hepatocellular carcinoma (HCC). We aimed to elucidate the correlation of PVT1 with hepatitis B virus-positive HCC in the clinic, and the roles of PVT1 in liver cancer cell biology, as well as to investigate the underlying molecular mechanisms. qPCR analysis was performed to examine the expression of PVT1 in hepatitis B virus-positive HCC tissues and liver cancer cell lines. lncRNA PVT1 overexpression and knockdown were achieved by transfection of an overexpression vector or shRNA. Cell proliferation, colony formation, migration, apoptosis, and invasion capabilities were examined, accordingly. RNA pull-down assay was employed to examine the connection between PVT1 and the PRC2 complex. Chromatin immunoprecipitation was employed to test the combination with EZH2 protein and H3K27me3 level on the MYC promoter. The results revealed that upregulation of PVT1 was detected in hepatitis B virus-positive HCC tissues compared with that noted in the HBV-negative samples. lncRNA PVT1 enhanced cell proliferation, migration, and invasion in the hepatitis B virus-positive Hep3B cells rather than the hepatitis B virus-negative HepG2 cells. PVT1 was able to bind EZH2 and obstruct the recruitment of EZH2 to the promoter of MYC therefore promoting MYC expression by altering H3K37me3 status in Hep3B liver cancer cells, and EZH2 protein was negatively correlated with lncRNA-PVT1 expression. In conclusion, our results indicate that lncRNA PVT1 promotes hepatitis B virus-positive liver cancer progression by disturbing histone methylation on the MYC promoter, suggesting that lncRNA PVT1 may be a potential target for developing diagnostic and therapeutic strategies of hepatitis B virus-positive liver cancer at the early stages.