Hepatitis B core antigen modulates exosomal miR-135a to target vesicle-associated membrane protein 2 promoting chemoresistance in hepatocellular carcinoma.
Hepatitis B core antigen modulates exosomal miR-135a to target vesicle-associated membrane protein 2 promoting chemoresistance in hepatocellular carcinoma.
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乙型肝炎核心抗原调节外泌体 miR-135a 以靶向囊泡相关膜蛋白 2，促进肝细胞癌的化疗耐药性
DOI：
10.3748/wjg.v27.i48.8302
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发表时间：
2021-12-28
影响因子：
4.3
通讯作者：
Zhu F
中科院分区：
文献类型：
--
作者：
Wei XC;Xia YR;Zhou P;Xue X;Ding S;Liu LJ;Zhu F
背景肝细胞癌（Hepatocellular carcinoma，HCC）是最常见的恶性肿瘤之一。B型肝炎病毒（HBV）感染与肝细胞癌的关系迄今已有文献报道。外泌体miRNA有助于癌症进展和化学抗性。已知HBV X蛋白调节促进细胞增殖和肝癌发生过程的miRNA。然而，关于B型肝炎核心抗原（HBc）调控exosomal miRNAs诱导肝癌细胞耐药的研究尚未见报道。目的探讨HBc促进肝癌细胞对阿霉素耐药的机制。方法超离心法分离外泌体。通过动态光散射（DLS）和透射电子显微镜（TEM）评价外泌体的形态和大小。使用癌症基因组图谱（TCGA）数据库鉴定HCC中差异表达的miRNA。采用定量聚合酶链反应检测患者组织标本中miR-135 a-5 p的水平。利用TargetScan和荧光素酶法对miR-135 a-5 p的靶基因进行预测和验证。最后，我们采用流式细胞术、CCK-8法和western blot法检测了miR-135 a-5 p在Dox存在或不存在的情况下对肝癌细胞增殖和抗凋亡的影响。结果发现HBc可增加miR-135 a-5 p的表达。生物信息学和患者样本的综合分析发现，与癌旁组织相比，miR-135 a-5 p在HCC组织中增加。生物信息学分析和体外验证将囊泡相关膜蛋白2（VAMP 2）鉴定为miR-135 a-5 p的新靶基因。功能分析显示，外泌体miR-135 a-5 p诱导HCC中的凋亡保护、细胞增殖和化疗抗性。此外，拯救实验表明，VAMP 2逆转了miR-135 a-5 p的凋亡保护、细胞生长和耐药性。最后，HBc通过miR-135 a-5 p/VAMP 2轴促进HCC抗凋亡、增殖和耐药性，并阻止Dox诱导的凋亡。结论HBc通过上调外泌体miR-135 a-5 p/VAMP 2的表达，促进细胞抗凋亡、增殖和耐药。因此，我们的工作表明miR-135 a-5 p/VAMP 2调节轴在HCC的化疗耐药性中起重要作用，并且是HCC的潜在分子治疗靶点。
BACKGROUND Hepatocellular carcinoma (HCC) is one of the most common malignant tumors. The association of hepatitis B virus (HBV) infection with HCC is hitherto documented. Exosomal miRNAs contribute to cancer progression and chemoresistance. HBV X protein has been known to modulate miRNAs that facilitate cell proliferation and the process of hepatocarcinogenesis. However, there has been no report on hepatitis B core antigen (HBc) regulating exosomal miRNAs to induce drug resistance of HCC cells. AIM To elucidate the mechanism by which HBc promotes Doxorubicin hydrochloride (Dox) resistance in HCC. METHODS Exosomes were isolated by ultracentrifugation. The morphology and size of exosomes were evaluated by Dynamic Light Scattering (DLS) and transmission electron microscopy (TEM). The miRNAs differentially expressed in HCC were identified using The Cancer Genome Atlas (TCGA) database. The level of miR-135a-5p in patient tissue samples was detected by quantitative polymerase chain reaction. TargetScan and luciferase assay were used to predict and prove the target gene of miR-135a-5p. Finally, we identified the effects of miR-135a-5p on anti-apoptosis and the proliferation of HCC in the presence or absence of Dox using flow cytometry, Cell counting kit 8 (CCK-8) assay and western blot. RESULTS We found that HBc increased the expression of exosomal miR-135a-5p. Integrated analysis of bioinformatics and patient samples found that miR-135a-5p was increased in HCC tissues in comparison with paracancerous tissues. Bioinformatic analysis and in vitro validation identified vesicle-associated membrane protein 2 (VAMP2) as a novel target gene of miR-135a-5p. Functional assays showed that exosomal miR-135a-5p induced apoptosis protection, cell proliferation, and chemotherapy resistance in HCC. In addition, the rescue experiment demonstrated that VAMP2 reversed apoptosis protection, cell growth, and drug resistance by miR-135a-5p. Finally, HBc promoted HCC anti-apoptosis, proliferation, and drug resistance and prevented Dox-induced apoptosis via the miR-135a-5p/VAMP2 axis. CONCLUSION These data suggested that HBc upregulated the expression of exosomal miR-135a-5p and promoted anti-apoptosis, cell proliferation, and chemical resistance through miR-135a-5p/VAMP2. Thus, our work indicated an essential role of the miR-135a-5p/VAMP2 regulatory axis in chemotherapy resistance of HCC and a potential molecular therapeutic target for HCC.
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DOI：
10.1073/pnas.1521230113
发表时间：
2016-02-23
影响因子：
11.1
作者：
Kowal, Joanna;Arras, Guillaume;Thery, Clotilde
通讯作者：
Thery, Clotilde
影响因子：
5.6
作者：
Kouwaki T;Okamoto M;Tsukamoto H;Fukushima Y;Oshiumi H
通讯作者：
Oshiumi H
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DOI：
10.1186/s13046-018-0677-7
发表时间：
2018-03-12
期刊：
Journal of experimental & clinical cancer research : CR
影响因子：
--
作者：
Fu X;Liu M;Qu S;Ma J;Zhang Y;Shi T;Wen H;Yang Y;Wang S;Wang J;Nan K;Yao Y;Tian T
通讯作者：
Tian T
影响因子：
14.9
作者：
Karagkouni D;Paraskevopoulou MD;Chatzopoulos S;Vlachos IS;Tastsoglou S;Kanellos I;Papadimitriou D;Kavakiotis I;Maniou S;Skoufos G;Vergoulis T;Dalamagas T;Hatzigeorgiou AG
通讯作者：
Hatzigeorgiou AG
乙型肝炎病毒核心蛋白以c-Ets2依赖性方式增强人端粒酶逆转录酶表达和肝癌细胞增殖
DOI：
10.1016/j.biocel.2013.03.015
发表时间：
2013-07-01
影响因子：
4
作者：
Gai, Xiaoxiao;Zhao, Peiqing;Liang, Xiaohong
通讯作者：
Liang, Xiaohong