Hepatitis B Virus Surface Antigen Promotes Stemness of Hepatocellular Carcinoma through Regulating MicroRNA-203a.
Hepatitis B Virus Surface Antigen Promotes Stemness of Hepatocellular Carcinoma through Regulating MicroRNA-203a.
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DOI：
10.14218/jcth.2021.00373
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发表时间：
2023-02-28
影响因子：
3.6
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中科院分区：
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持续阳性的B型肝炎表面抗原（HBsAg）患者，即使HBV-DNA载量较低，发生肝细胞癌（HCC）的风险也高于无HBV感染的患者。鉴于肿瘤的干性在肿瘤的发生和维持中具有关键作用，本研究旨在探讨HBsAg是否通过调控microRNA影响HCC的生物学功能和干性，并探讨其潜在机制。我们筛选出了miR-203 a，这是在对HBsAg阳性样本的微阵列分析中最显著下调的microRNA，并在随后的研究中专注于该miRNA。进行体外和体内功能实验以评估其调节功能。本研究分析了miR-203 a对干性的影响以及与BMI 1的可能相关性。miR-203 a在HBeAg阳性HCC中显著下调，微阵列分析中显示最急剧的降低。在细胞中刺激或过表达/敲低HBsAg后，通过定量实时PCR证实miR-203 a与HBsAg表达之间的负相关性。我们证明了miR-203 a在体内抑制HCC细胞增殖、迁移、克隆形成能力和肿瘤发展中的功能。此外，miR-203 a的过表达显著增加了肿瘤细胞对5-FU治疗的敏感性，并降低了具有干细胞标志物的HCC细胞的比例。与我们的研究一致，癌症基因组图谱数据库和我们中心的样本的生存分析表明，miR-203 a水平低的患者预后较差。我们还发现维持干细胞自我更新能力的基因BMI 1与HCC标本中的miR-203 a呈显著负相关（p<0.001）。类似地，也在体外证实了miR-203 a过表达/敲低后的相反BMI 1变化。双荧光素酶报告分析表明miR-203 a可能通过直接结合调控BMI 1的表达。HBsAg可能通过抑制miR-203 a促进HCC的发展和肿瘤的干化，导致预后不良。miR-203 a可能是治疗HCC的重要靶点。下一步需要进行更明确的机制研究和动物实验。
Patients with persistent positive hepatitis B surface antigen (HBsAg), even with a low HBV-DNA load, have a higher risk of hepatocellular carcinoma (HCC) than those without HBV infection. Given that tumor stemness has a critical role in the occurrence and maintenance of neoplasms, this study aimed to explore whether HBsAg affects biological function and stemness of HCC by regulating microRNA, and to explore underlying mechanisms. We screened out miR-203a, the most significant down-regulated microRNA in the microarray analysis of HBsAg-positive samples and focused on that miRNA in the ensuing study. In vitro and in vivo functional experiments were performed to assess its regulatory function. The effect of miR-203a on stemness and the possible correlation with BMI1 were analyzed in this study. MiR-203a was significantly down-regulated in HBsAg-positive HCC with the sharpest decrease shown in microarray analysis. The negative correlation between miR-203a and HBsAg expression was confirmed by quantitative real-time PCR after stimulation or overexpression/knockdown of HBsAg in cells. We demonstrated the function of miR-203a in inhibiting HCC cell proliferation, migration, clonogenic capacity, and tumor development in vivo. Furthermore, the overexpression of miR-203a remarkably increases the sensitivity of tumor cells to 5-FU treatment and decreases the proportion of HCC cells with stem markers. In concordance with our study, the survival analysis of both The Cancer Genome Atlas database and samples in our center indicated a worse prognosis in patients with low level of miR-203a. We also found that BMI1, a gene maintains the self-renewal capacity of stem cells, showed a significant negative correlation with miR-203a in HCC specimen (p<0.001). Similarly, opposite BMI1 changes after overexpression/knockdown of miR-203a were also confirmed in vitro. Dual luciferase reporting assay suggested that miR-203a may regulate BMI1 expression by direct binding. HBsAg may promote the development of HCC and tumor stemness by inhibiting miR-203a, resulting in poor prognosis. miR-203a may serve as a crucial treatment target in HBsAg-positive HCC. More explicit mechanistic studies and animal experiments need to be conducted as a next step.