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1.
J Vis Exp ; (144)2019 02 19.
Artículo en Inglés | MEDLINE | ID: mdl-30855577

RESUMEN

Despite the exceptional infectivity of the hepatitis B virus (HBV) in vivo, where only three viral genomes can result in a chronicity of experimentally infected chimpanzees, most in vitro models require several hundreds to thousands of viral genomes per cell in order to initiate a transient infection. Additionally, static 2D cultures of primary human hepatocytes (PHH) allow only short-term studies due to their rapid dedifferentiation. Here, we describe 3D liver-on-a-chip cultures of PHH, either in monocultures or in cocultures with other nonparenchymal liver-resident cells. These offer a significant improvement to studying long-term HBV infections with physiological host cell responses. In addition to facilitating drug efficacy studies, toxicological analysis, and investigations into pathogenesis, these microfluidic culture systems enable the evaluation of curative therapies for HBV infection aimed at eliminating covalently closed, circular (ccc)DNA. This presented method describes the set-up of PHH monocultures and PHH/Kupffer cell co-cultures, their infection with purified HBV, and the analysis of host responses. This method is particularly applicable to the evaluation of long-term effects of HBV infection, treatment combinations, and pathogenesis.


Asunto(s)
Virus de la Hepatitis B/fisiología , Hepatitis B/fisiopatología , Hepatocitos/metabolismo , Macrófagos del Hígado/metabolismo , Hígado/patología , Humanos
2.
Data Brief ; 14: 538-542, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28856185

RESUMEN

This data contains information related to the research article entitled "Osteopontin splice variants and polymorphisms in Cancer Progression and Prognosis" [1]. Here, we describe an in silico analysis of transcription factors that could have altered binding to their DNA target sequence as a result of SNPs in the osteopontin gene promoter. We concentrated on SNPs associated with cancer risk and development. The analysis was performed with PROMO v3.0.2 software which incorporates TRANSFACT v6.4 of. We also present a figure depicting the putative transcription factor binding according to genotype.

3.
Biochim Biophys Acta Rev Cancer ; 1868(1): 93-108.A, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28254527

RESUMEN

Osteopontin (OPN) is an extracellular matrix protein that is overexpressed in various cancers and promotes oncogenic features including cell proliferation, survival, migration, and angiogenesis, among others. OPN can participate in the regulation of the tumor microenvironment, affecting both cancer and neighboring cells. Here, we review the roles of OPN splice variants (a, b, c) in cancer development, progression, and prognosis, and also discuss the identities of isoforms 4 and 5. We also discussed how single-nucleotide polymorphisms (SNPs) of the OPN gene are an additional factor influencing the level of OPN in individuals, modulating the risks of cancer development and outcome.


Asunto(s)
Neoplasias/genética , Neoplasias/patología , Osteopontina/genética , Polimorfismo Genético/genética , Empalme del ARN/genética , Secuencia de Aminoácidos , Secuencia de Bases , Progresión de la Enfermedad , Humanos , Pronóstico
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