miRNA-122表达的乙肝病毒X蛋白介导的抑制通过细胞周期G1-P53轴

StephenW

Infect Agent Cancer. 2016 Aug 15;11:40. doi: 10.1186/s13027-016-0085-6. eCollection 2016.
Hepatitis B virus X protein mediated suppression of miRNA-122 expression enhances hepatoblastoma cell proliferation through cyclin G1-p53 axis.Bandopadhyay M1, Sarkar N1, Datta S2, Das D1, Pal A1, Panigrahi R3, Banerjee A1, Panda CK4, Das C5, Chakrabarti S6, Chakravarty R1.
Author information

• 1ICMR Virus Unit, Kolkata, Indian Council of Medical Research, GB-4, 1st floor, ID & BG Hospital Campus, 57, Dr. S C Banerjee Road, Beliaghata, Kolkata, 700010 West Bengal India.
• 2Molecular Virology Laboratory, Defense Research Laboratory (DRDO), Tezpur, Assam India.
• 3ICMR Virus Unit, Kolkata, Indian Council of Medical Research, GB-4, 1st floor, ID & BG Hospital Campus, 57, Dr. S C Banerjee Road, Beliaghata, Kolkata, 700010 West Bengal India ; Present Address: Department of Pathology & Lab Medicine, Tulane University School of Medicine, New Orleans, LA 70112 USA.
• 4Chittaranjan National Cancer Institute, 37, SP Mukherjee Road, Kolkata, India.
• 5Saha Institute of Nuclear Physics, Bidhan nagar, Kolkata India.
• 6National Institute of Cholera and Enteric Diseases, Kolkata, India.
  


AbstractBACKGROUND: Hepatitis B virus (HBV) X protein (HBx) reported to be associated with pathogenesis of hepatocellular carcinoma (HCC) and miR-122 expression is down regulated in HCC. Previous studies reported miR-122 targets cyclin G1 (CCNG1) expression and this in turn abolishes p53-mediated inhibition of HBV replication. Here we investigated the involvement of HBx protein in the modulation of miR-122 expression in hepatoblastoma cells.
METHODS: Expression of miR-122 was measured in HepG2 cells transfected with HBx plasmid (HBx-HepG2), full length HBV genome (HBV-HepG2) and in constitutively HBV synthesizing HepG2.2.15 cells. CCNG1 mRNA (a direct target of miR-122) and protein expressions were also measured in both HBx-HepG2, HBV-HepG2 cells and in HepG2.2.15 cells. miR-122 expressions were analyzed in HBx-HepG2, HBV-HepG2 and in HepG2.2.15 cells after treatment with HBx mRNA specific siRNA. Expressions of p53 mRNA and protein which is negatively regulated by CCNG1 were analyzed in HBx transfected HepG2 cells; X silenced HBx-HepG2 cells and X silenced HepG2.2.15 cells. HBx induced cell proliferation in HepG2 cells was measured by cell proliferation assay. Flow cytometry was used to evaluate changes in cell cycle distribution. Expression of cell cycle markers were measured by real time PCR.
RESULTS: Expression of miR-122 was down regulated in HBx-HepG2, HBV-HepG2 and also in HepG2.2.15 cell line compared to control HepG2 cells. CCNG1 expression was found to be up regulated in HBx-HepG2, HBV-HepG2 cells and in HepG2.2.15 cells. Following siRNA mediated silencing of HBx expression; increased miR-122 levels were documented in HBx-HepG2, HBV-HepG2 and in HepG2.2.15 cells. HBx silencing in HBx-HepG2 and HepG2.2.15 cells also resulted in increased p53 expression. FACS analysis and assessment of expressions of cell cycle markers revealed HBx induced a release from G1/S arrest in HepG2 cells. Further, cell proliferation assay showed HBx promoted proliferation of HepG2 cell.
CONCLUSION: Our study revealed that HBx induced down regulation of miR-122 expression that consequently increased CCNG1 expression. This subsequently caused cell proliferation and release from G1/S arrest in malignant hepatocytes. The study provides the potential to utilize the HBx-miR-122 interaction as a therapeutic target to limit the development of HBV related HCC.


KEYWORDS: CCNG1; HBx; HepG2; HepG2.2.15; Hepatocellular carcinoma; miR-122

PMID:27528885PMCID:PMC4983788DOI:10.1186/s13027-016-0085-6
[PubMed] Free PMC Article

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StephenW

感染癌症代理。 2016年8月15日，11：40。 DOI：10.1186 / s13027-016-0085-6。 eCollection 2016年。
的miRNA-122表达的乙肝病毒X蛋白介导的抑制通过细胞周期G1-P53轴增强肝母细胞瘤细胞增殖。
Bandopadhyay M1，N1萨卡，达塔S2，达斯D1，帕尔A1，Panigrahi R3，班纳吉A1，熊猫CK4，达斯C5，查克拉巴蒂S6，R1 Chakravarty。
作者信息

    1ICMR病毒科，加尔各答，医学研究印第安人理事会，GB-4，一楼，ID＆BG医院校区，57，了C班纳吉博士路，Beliaghata，加尔各答，700010西孟加拉邦印度。
    2Molecular病毒学实验室，国防研究实验室（DRDO），提斯普尔，印度阿萨姆邦。
    3ICMR病毒科，加尔各答，医学研究印第安人理事会，GB-4，一楼，ID＆BG医院校区，57，了C班纳吉博士路，Beliaghata，加尔各答，700010西孟加拉邦印度;现住址：病理学与实验医学，医学杜兰大学医学院，新奥尔良，LA 70112 USA系。
    4Chittaranjan国家癌症研究所，37，SP慕克吉路，加尔各答，印度。
    核物理研究所5Saha，比丹纳加尔，加尔各答印度。
    霍乱和肠道疾病，加尔各答，印度6National研究所。

抽象
背景：

乙型肝炎病毒（HBV）X蛋白（HBx蛋白）报道与肝细胞癌（HCC）和miR-122表达的发病机制在HCC中下调相关联。先前的研究报道的miR-122靶向细胞周期蛋白G1（CCNG1）的表达，这反过来废除HBV复制的p53介导的抑制作用。这里，我们调查的HBx蛋白在的miR-122表达在肝母细胞瘤细胞中的调制的参与。
方法：

与HBx的质粒（HBx蛋白-HepG2细胞）转染HepG2细胞，全长HBV基因组（HBV-HepG2细胞）和HBV组成合成HepG2.2.15细胞测定的miR-122的表达。 CCNG1 mRNA和蛋白表达（MIR-122的直接靶）在这两个HBx蛋白，肝癌，HBV-HepG2细胞和HepG2.2.15细胞进行了测定。的miR-122表达的HBx蛋白-HepG2细胞，HBV-HepG2和在HepG2.2.15细胞与HBx的mRNA的特异性siRNA处理后进行分析。其负面CCNG1中的HBx分析调节p53的mRNA和蛋白表达的转染的HepG2细胞;点¯x沉默HBx蛋白，HepG2细胞和X沉默HepG2.2.15细胞。在HepG2细胞中的HBx诱导的细胞增殖通过细胞增殖测定来测量。流式细胞术用来评估在细胞周期分布的改变。通过实时定量PCR检测的细胞周期标志物的表达。
结果：

的miR-122的表达HBx蛋白，肝癌，HBV-HepG2细胞下调，也HepG2.2.15细胞的细胞系相比，控制HepG2细胞。发现CCNG1表达在HBx蛋白，肝癌，HBV-HepG2细胞和HepG2.2.15细胞被上调。继HBx蛋白表达的siRNA介导的沉默;增加的miR-122水平HBx蛋白，肝癌，HBV-HepG2细胞和HepG2.2.15细胞被记录在案。在HBx蛋白，HepG2和HepG2.2.15细胞HBx基因沉默也导致增加p53的表达。 FACS分析和细胞周期标志物表达的评估表明HBx蛋白诱导从G1 / S阻滞HepG2细胞释放。此外，细胞增殖实验显示HBx蛋白促进肝癌细胞增殖。
结论：

我们的研究表明，HBx蛋白诱导下调的miR-122表达了随之增加CCNG1表达的调控。随后这导致细胞增殖和从G1 / S阻滞在恶性肝细胞释放。该研究提供了利用的HBx-的miR-122相互作用作为治疗靶，以限制HBV相关HCC的发展潜力。
关键词：

CCNG1; HBx蛋白; HepG2细胞; HepG2.2.15细胞;肝细胞癌;的miR-122

结论：
    27528885
PMCID：
    PMC4983788
DOI：
    10.1186 / s13027-016-0085-6

    [考研]

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