Shi Taiyang, Hua Qinfang, Ma Zhipeng, Lv Qijun
Department of Laboratory Medicine, Central Laboratory, Shengli Oilfield Central Hospital, Dongying 257034, Shandong, China.
Department of Pharmacy, People's Hospital of Hekou District, Dongying 257200, Shandong, China.
Pathol Res Pract. 2017 Dec;213(12):1464-1469. doi: 10.1016/j.prp.2017.10.020. Epub 2017 Oct 26.
Hepatitis B Virus X (HBx) Protein encoded by HBV is believed to be the major player in the process of HBV-induced oncogenesis. Ectopic expression of miR-200a-3p was reported to be associated with diverse tumorigenesis. This study aimed to better understand the role of miR-200a-3p and its correlation with HBx in HBV-induced hepatocellular carcinoma (HCC).
In this report, we examined the gene expression using quantitative RT-PCR and protein expression using Western blotting analysis. Cells were transfected with miR-200a-3p mimics or empty vector, and HBx-carrying vector or empty vector. Cell viability was tested using CCK-8 assay. Wound healing assay was performed to assess cell migration while Transwell assay was performed to evaluate cell invasion.
miR-200a-3p was downregulated in HBV-positive tissue samples compared with HBV-negative tissue samples. This result was further confirmed with HBV-positive and - negative cell lines. HBx protein was overexpressed in HBV-positive cells where expression of miR-200a-3p was significantly suppressed. Increased cell viability, altered cell cycle progression, increased cell migration and invasion occurred in HBx-overexpressed cells compared to its controls. In forced expressed miR-200a-3p cells, cell viability, cell migration and invasion were significantly decreased, and cell cycle status was altered compared to its controls.
Taken together, pathogenetic function of HBx is negatively correlated with miR-200a-3p in HBV-cased HCC through regulating cell viability, cell cycle arrest, cell migration and cell invasion.
乙肝病毒（HBV）编码的乙肝病毒X蛋白（HBx）被认为是HBV诱导肿瘤发生过程中的主要参与者。据报道，miR-200a-3p的异位表达与多种肿瘤发生有关。本研究旨在更好地了解miR-200a-3p的作用及其与HBx在HBV诱导的肝细胞癌（HCC）中的相关性。
在本报告中，我们使用定量RT-PCR检测基因表达，并使用蛋白质印迹分析检测蛋白质表达。用miR-200a-3p模拟物或空载体以及携带HBx的载体或空载体转染细胞。使用CCK-8测定法测试细胞活力。进行伤口愈合试验以评估细胞迁移，同时进行Transwell试验以评估细胞侵袭。
与HBV阴性组织样本相比，HBV阳性组织样本中miR-200a-3p表达下调。HBV阳性和阴性细胞系进一步证实了这一结果。HBx蛋白在miR-200a-3p表达明显受抑制的HBV阳性细胞中过表达。与对照相比，HBx过表达的细胞中细胞活力增加、细胞周期进程改变、细胞迁移和侵袭增加。在强制表达miR-200a-3p的细胞中，与对照相比，细胞活力、细胞迁移和侵袭明显降低，并且细胞周期状态发生改变。
综上所述，在HBV相关的HCC中，HBx的致病功能通过调节细胞活力、细胞周期停滞、细胞迁移和细胞侵袭与miR-200a-3p呈负相关。
