miR-325-3p/DPAGT1 轴失调促进 HBV 阳性 HCC 化疗耐药。
Dysregulation of the miR-325-3p/DPAGT1 axis supports HBV-positive HCC chemoresistance.
机构信息
Department of Immunology, Anhui Key Laboratory of Infection and Immunity at Bengbu Medical College, Bengbu, Anhui, China.
Department of Laboratory Medicine, Bengbu Medical College, Bengbu, Anhui, China.
出版信息
Biochem Biophys Res Commun. 2019 Nov 5;519(2):358-365. doi: 10.1016/j.bbrc.2019.08.116. Epub 2019 Sep 10.
BACKGROUND
Chemotherapeutic resistance in hepatitis B virus (HBV)-positive hepatocellular carcinoma (HCC) patients is an unfortunate side effect of standard chemotherapy. This situation necessitates a better understanding of the molecular pathways underlying HBV + HCC chemoresistance in order to aid the development of novel chemotherapeutic targets.
METHODS
We generated two doxorubicin (DOX)-resistant HBV + HCC sublines HepG2.2.15 and Huh7-1.3. qRT-PCR was used to evaluate dysregulation in hexosamine pathway genes in chemosensitive and chemoresistant HBV + HCC cell lines in vitro. Western blots, luciferase reporter assays, and in vivo xenograft tumor studies were conducted to reveal the role of the miRNA-325-3p/DPAGT1 axis in HBV + HCC chemoresistance.
RESULTS
The hexosamine pathway gene dolichyl-phosphate N-acetylglucosamine phosphotransferase 1 (DPAGT1) was found to be upregulated in both DOX-resistant cell lines. Enhancing DPAGT1 activity significantly improved the survival of DOX-resistant cells. Silencing or pharmacological inhibition of DPAGT1 inhibited xenograft tumor growth under DOX-treated conditions. DPAGT1 upregulation was associated with higher levels of stemness-related markers and ATP-binding cassette (ABC) drug efflux transporters in DOX-resistant cell lines. miR-325-3p was found to negatively modulate DPAGT1 expression and phenocopied the effects of DPAGT1 silencing in vitro and in vivo. In HBV + HCC patients treated with transarterial chemoembolization (TACE), high and low levels of tumor DPAGT1 and miR-325-3p expression, respectively, were associated with a poor chemotherapeutic response.
CONCLUSIONS
Our findings provide novel insights into the role of miR-325-3p/DPAGT1 axis dysregulation in supporting HBV + HCC chemoresistance.
背景
乙型肝炎病毒（HBV）阳性肝细胞癌（HCC）患者的化疗耐药是标准化疗的不幸副作用。这种情况需要更好地了解 HBV+HCC 化疗耐药的分子途径，以帮助开发新的化疗靶点。
方法
我们生成了两种阿霉素（DOX）耐药 HBV+HCC 亚系 HepG2.2.15 和 Huh7-1.3。体外 qRT-PCR 用于评估化学敏感性和耐药性 HBV+HCC 细胞系中己糖胺途径基因的失调。Western blot、荧光素酶报告基因检测和体内异种移植肿瘤研究揭示了 miRNA-325-3p/DPAGT1 轴在 HBV+HCC 化疗耐药中的作用。
结果
己糖胺途径基因 dolichyl-phosphate N-acetylglucosamine phosphotransferase 1（DPAGT1）在两种 DOX 耐药细胞系中均上调。增强 DPAGT1 活性可显著提高 DOX 耐药细胞的存活率。沉默或药理学抑制 DPAGT1 可抑制 DOX 处理条件下的异种移植肿瘤生长。DPAGT1 上调与 DOX 耐药细胞系中更高水平的干性相关标志物和三磷酸腺苷结合盒（ABC）药物外排转运体相关。miR-325-3p 被发现可负调控 DPAGT1 表达，并在体外和体内模拟 DPAGT1 沉默的作用。在接受经动脉化疗栓塞（TACE）治疗的 HBV+HCC 患者中，肿瘤 DPAGT1 和 miR-325-3p 表达水平高和低分别与化疗反应不良相关。
结论
我们的研究结果提供了新的见解，即 miR-325-3p/DPAGT1 轴失调在支持 HBV+HCC 化疗耐药中的作用。
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