Cancer remains one of the main causes of human mortality despite significant progress in its diagnostics and therapy achieved in the past decade. Massive hypomethylation of retrotransposons, in particular LINE-1, is considered a hallmark of most malignant transformations as it results in the reactivation of retroelements and subsequent genomic instability. Accumulating data on LINE-1 aberrant methylation in different tumor types indicates its significant role in cancer initiation and progression. However, direct evidence that LINE-1 activation can be used as a cancer biomarker is still limited. The objective of this review was to critically evaluate the published results regarding the diagnostic/prognostic potential of the LINE-1 methylation status in cancer. Our analysis indicates that LINE-1 hypomethylation is a promising candidate biomarker of cancer development, which, however, needs validation in both clinical and laboratory studies to confirm its applicability to different cancer types and/or stages. As LINE-1 is present in multiple cell-free copies in blood, it has advantages over single-copy genes regarding perspectives of using its methylation status as an epigenetic cancer biomarker for cell-free DNA liquid biopsy.

• Keywords
• DNA methylation, LINE-1 (L1), cell-free DNA, epigenetic cancer biomarker,
• MeSH
• Survival Analysis MeSH
• Antineoplastic Agents therapeutic use   MeSH
• Long Interspersed Nucleotide Elements * MeSH
• Epigenesis, Genetic MeSH
• Humans MeSH
• DNA Methylation MeSH
• Biomarkers, Tumor blood   genetics   MeSH
• Neoplasms diagnosis   drug therapy   genetics   mortality   MeSH
• Genomic Instability MeSH
• Prognosis MeSH
• Disease Progression MeSH
• Gene Expression Regulation, Neoplastic * MeSH
• Signal Transduction MeSH
• Liquid Biopsy MeSH
• DNA Transposable Elements * MeSH
• Cell-Free Nucleic Acids blood   genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• Antineoplastic Agents MeSH
• Biomarkers, Tumor MeSH
• DNA Transposable Elements * MeSH
• Cell-Free Nucleic Acids MeSH

The patatin-like phospholipase domain containing 3 (PNPLA3) gene (viz. its I148M variant) is one of the key players in the pathogenesis of nonalcoholic fatty liver disease (NAFLD). We have identified a novel insertion/deletion variant of 1114 bp, localized in the second intron of the PNPLA3 gene, which corresponds to the 3' terminal sequence of the long-interspersed element (LINE-1). DNA analysis of 122 NAFLD patients and 167 control subjects as well as RNA analysis of 19 liver biopsies revealed that the novel variant is very common (frequency = 0.41), fully linked to the clinically important I148M variant, and clinically silent. Although the LINE-1 insertion does not seem to have any biological effect, it can impede genotyping of the I148M variant. If insertion prevents the attachment of the diagnostic primer, then the non-insertion allele will be selectively amplified; and thus the frequency of the 148M "risk" allele will be significantly overestimated due to the complete linkage of the LINE-1 insertion and the 148I allele of the PNPLA3 gene. Therefore, our findings underline the importance of careful design and consistent documentation of the methodology, including primer sequences. Critical revisions of the results of some studies that have already been reported may therefore be needed.

• MeSH
• Acyltransferases genetics   MeSH
• Alleles MeSH
• Long Interspersed Nucleotide Elements genetics   MeSH
• Phospholipases A2, Calcium-Independent genetics   MeSH
• Genetic Predisposition to Disease genetics   MeSH
• Genotype MeSH
• Liver pathology   MeSH
• Polymorphism, Single Nucleotide genetics   MeSH
• Humans MeSH
• Non-alcoholic Fatty Liver Disease genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Acyltransferases MeSH
• adiponutrin, human MeSH Browser
• Phospholipases A2, Calcium-Independent MeSH

We describe the molecular etiology of β(+)-thalassemia that is caused by the insertion of the full-length transposable element LINE-1 (L1) into the intron-2 of the β-globin gene (HBB). The transcript level of the affected β-globin gene was severely reduced. The remaining transcripts consisted of full-length, correctly processed β-globin mRNA and a minute amount of three aberrantly spliced transcripts with a decreased half-life due to activation of the nonsense-mediated decay pathway. The lower steady-state amount of mRNA produced by the β-globin(L1) allele also resulted from a reduced rate of transcription and decreased production of full-length β-globin primary transcripts. The promoter and enhancer sequences of the β-globin(L1) allele were hypermethylated; however, treatment with a demethylating agent did not restore the impaired transcription. A histone deacetylase inhibitor partially reactivated the β-globin(L1) transcription despite permanent β-globin(L1) promoter CpG methylation. This result indicates that the decreased rate of transcription from the β-globin(L1) allele is associated with an altered chromatin structure. Therefore, the molecular defect caused by intronic L1 insertion in the β-globin gene represents a novel etiology of β-thalassemia.

• Keywords
• HBB, LINE-1, epigenetic repression, β-globin, β-thalassemia,
• MeSH
• Alleles MeSH
• Alternative Splicing MeSH
• beta-Globins genetics   MeSH
• beta-Thalassemia genetics   MeSH
• CpG Islands MeSH
• Long Interspersed Nucleotide Elements * MeSH
• Adult MeSH
• Transcription, Genetic MeSH
• Introns * MeSH
• Mutagenesis, Insertional * MeSH
• Humans MeSH
• DNA Methylation MeSH
• Gene Order MeSH
• Promoter Regions, Genetic MeSH
• Gene Expression Regulation MeSH
• RNA Stability MeSH
• Gene Silencing MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Case Reports MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• beta-Globins MeSH

BACKGROUND: Neurotensin receptors are overexpressed in several cancer types including pancreatic ductal adenocarcinoma. Three NTR subtypes have been cloned: NTR-1, NTR-2 and NTR-3. The most expressed NTR-1 is not present in normal pancreatic tissue and has a low expression in chronic pancreatitis. OBJECTIVE: Objective of this study was to test in vitro affinity of the new 68Ga labelled neurotensin analogue DOTA-NT-20.3 (fragment 6-13, Ac-Lys(DOTA)-Pro-Arg(N-CH3)-Arg-Pro-Tyr-Tle-Leu) on the human pancreatic ductal adenocarcinoma cell line AsPC-1. METHOD: For the preparation of 68Ga-DOTA-NT-20.3, 68GaCl3 solution (eluted from 68Ge/68Ga generator) and 50 μg of precursor (Iason, Graz, Austria) water dissolved were used in an automatic synthesis module. The labeled compound was added to cell culture flask and incubated at 37°C. At various time points after tracer addition up to 80min, cells were recovered, rinsed and counted for radioactivity. Results were expressed as percent binding normalized to 200000 cells and affinity parameters were calculated. RESULTS: Labeling yield was ≥98 %. The molar ratio between labelled and total peptide was about 1/400. AsPC-1 cell line showed rapid uptake of the tracer including surface and internalized binding, tending to a plateau phase 80 min after tracer addition (11%/200.000 cells). The Kd (7.335 pmol) and Bmax (90.52 kBq) value indicated high tracer affinity for AsPC-1cell line especially if compared with the literature data regarding other malignancies (e.g. colonic cancer cell line). Binding sites were 1.09x106 sites per cell. CONCLUSION: New tracer 68Ga-DOTA-NT-20.3 can be a suitable candidate for the clinical use in patients with pancreatic ductal adenocarcinoma.

• Keywords
• 68Ga-NT-DOTA- 20.3, AsPC-1, NT, NTR-1, Neurotensin, Neurotensin receptors, Pancreatic ductal adenocarcinoma.,
• MeSH
• Adenocarcinoma metabolism   pathology   MeSH
• Heterocyclic Compounds, 1-Ring chemistry   metabolism   MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• Pancreatic Neoplasms metabolism   pathology   MeSH
• Neurotensin agonists   chemistry   metabolism   MeSH
• Peptide Fragments chemistry   metabolism   MeSH
• Gallium Radioisotopes chemistry   metabolism   MeSH
• Receptors, Neurotensin metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• 1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid MeSH Browser
• Heterocyclic Compounds, 1-Ring MeSH
• Neurotensin MeSH
• Peptide Fragments MeSH
• Gallium Radioisotopes MeSH
• Receptors, Neurotensin MeSH

We have found that pretreatment of a human oral mucosal epithelial cell line KB with natural human interferon-alpha (IFN-alpha) augments the expression of intercellular adhesion molecule-1 (ICAM-1), and the complex of CD29 and CD49b, which is known as very late antigen (VLA)-2, in a dose-dependent manner, whereas the expression of other adhesion molecules such as CD44 leukocyte function associated antigen-3 (LFA-3), VLA-4 and endothelial leukocyte adhesion molecule-1 (ELAM-1) did not show any significant changes under the same conditions. The pretreatment of KB cells with IFN-alpha also induces the adhesion of these cells to the human leukemia T cell line MOLT-16 and to peripheral T lymphocytes in a dose-dependent manner. However, the adhesion of the above-mentioned T cells to KB cells was not inhibited by specific antibodies against ICAM-1, CD29, and CD49b. The results thus show that adhesion molecules other than ICAM-1, CD29, AND CD49b are responsible for the induced adhesion between T cells and IFN-alpha-pretreated KB cells.

• MeSH
• Integrin beta1 biosynthesis   MeSH
• Cell Adhesion MeSH
• Cell Line MeSH
• Antigens, CD biosynthesis   MeSH
• Epithelium metabolism   MeSH
• Epithelial Cells MeSH
• Integrin alpha2 MeSH
• Interferon-alpha pharmacology   MeSH
• Humans MeSH
• Intercellular Adhesion Molecule-1 biosynthesis   MeSH
• T-Lymphocytes metabolism   pathology   MeSH
• Mouth Mucosa cytology   metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Integrin beta1 MeSH
• Antigens, CD MeSH
• Integrin alpha2 MeSH
• Interferon-alpha MeSH
• Intercellular Adhesion Molecule-1 MeSH

BACKGROUND: Human immunodeficiency virus type 1 (HIV-1) latency represents the major barrier to virus eradication in infected individuals because cells harboring latent HIV-1 provirus are not affected by current antiretroviral therapy (ART). We previously demonstrated that DNA methylation of HIV-1 long terminal repeat (5' LTR) restricts HIV-1 reactivation and, together with chromatin conformation, represents an important mechanism of HIV-1 latency maintenance. Here, we explored the new issue of temporal development of DNA methylation in latent HIV-1 5' LTR. RESULTS: In the Jurkat CD4(+) T cell model of latency, we showed that the stimulation of host cells contributed to de novo DNA methylation of the latent HIV-1 5' LTR sequences. Consecutive stimulations of model CD4(+) T cell line with TNF-α and PMA or with SAHA contributed to the progressive accumulation of 5' LTR DNA methylation. Further, we showed that once established, the high DNA methylation level of the latent 5' LTR in the cell line model was a stable epigenetic mark. Finally, we explored the development of 5' LTR DNA methylation in the latent reservoir of HIV-1-infected individuals who were treated with ART. We detected low levels of 5' LTR DNA methylation in the resting CD4(+) T cells of the group of patients who were treated for up to 3 years. However, after long-term ART, we observed an accumulation of 5' LTR DNA methylation in the latent reservoir. Importantly, within the latent reservoir of some long-term-treated individuals, we uncovered populations of proviral molecules with a high density of 5' LTR CpG methylation. CONCLUSIONS: Our data showed the presence of 5' LTR DNA methylation in the long-term reservoir of HIV-1-infected individuals and implied that the transient stimulation of cells harboring latent proviruses may contribute, at least in part, to the methylation of the HIV-1 promoter.

• Keywords
• Chromatin conformation, DNA methylation, HIV-1, HIV-1-infected individuals, Latent HIV-1 provirus reactivation, Latent reservoir,
• MeSH
• Cell Line virology   MeSH
• Time Factors MeSH
• HIV Long Terminal Repeat genetics   MeSH
• HIV Infections drug therapy   genetics   virology   MeSH
• HIV-1 genetics   MeSH
• Jurkat Cells virology   MeSH
• Virus Latency genetics   physiology   MeSH
• Anti-HIV Agents therapeutic use   MeSH
• Humans MeSH
• DNA Methylation * MeSH
• Proviruses genetics   physiology   MeSH
• Antiretroviral Therapy, Highly Active MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Anti-HIV Agents MeSH

Mitosis in spherical cells of heteronuclear cell line VUP-1 was analyzed by means of time-lapse cinemicrography. A comparison with another permanent cell line (hamster) revealed some differences in the duration of individual mitotic stages. Metaphase was the longest stage in VUP-1 line cells (about 3-4 hours). Various irregularities in mitotic cycle kinetics were also found, such as absence of anaphase or separation of chromosomes from the aster in metaphase. Attention was further focussed on cytokinesis.

• MeSH
• Cell Line * MeSH
• Epithelial Cells MeSH
• Fibroblasts MeSH
• Cricetinae MeSH
• Humans MeSH
• Melanoma MeSH
• Mitosis * MeSH
• Choroid Neoplasms MeSH
• Animals MeSH
• Check Tag
• Cricetinae MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Hybridization of established dendritic cell lines with tumour cells represents a prospective technology for the construction of antitumour vaccines. Experiments were designed to examine whether administration of cell populations prepared by fusion of HPV 16-associated tumour TC-1 cells with dendritic cell line DC2.4 could be used for treatment of TC-1 tumours growing in syngeneic mice. The therapeutic potency of TC-1/DC2.4 fusion vaccine administered 24 h after fusion and that of TC-1/DC2.4 hybrid cells selected for 3 weeks in HAT-containing medium was tested. It has been found that administration of both types of fusion vaccines at the site of growing TC-1 tumour transplants significantly inhibited tumour growth with regard to the percentage of tumour-bearing mice and to the size of the transplanted tumours. Peritumoral administration of the DC2.4 cells alone also reduced the size of growing TC-1 tumours, but not the percentage of the tumour-bearing mice. Although in the groups of mice treated with fusion vaccines the size of the tumours was reproducibly smaller than that in the mice treated with parental DC2.4 cells, the difference was not statistically significant.

• MeSH
• Dendritic Cells immunology   transplantation   MeSH
• Cell Fusion MeSH
• Hybrid Cells immunology   transplantation   MeSH
• Immunotherapy methods   MeSH
• Culture Media MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Cell Line, Tumor MeSH
• Neoplasm Proteins therapeutic use   MeSH
• Neoplasms, Glandular and Epithelial immunology   pathology   therapy   virology   MeSH
• Papillomaviridae physiology   MeSH
• Disease Progression MeSH
• Cancer Vaccines therapeutic use   MeSH
• Flow Cytometry MeSH
• Neoplasm Transplantation MeSH
• Papillomavirus Vaccines * MeSH
• Viral Vaccines therapeutic use   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Culture Media MeSH
• Neoplasm Proteins MeSH
• Cancer Vaccines MeSH
• TCIM protein, human MeSH Browser
• Papillomavirus Vaccines * MeSH
• Viral Vaccines MeSH

The HPRS line 1 lymphoblastoid cells derived from an ovarian lymphoma induced by Marek's disease virus in chickens displayed increased agglutinability and haemadsorption when incubated with Con A. The increase was proportional to the concentration of Con A used. The same Con A-untreated cells and normal chicken lymphocytes incubated with Con A exhibited no increase in agglutinability and haemadsorption, however.

• MeSH
• Agglutination drug effects   MeSH
• Cell Membrane drug effects   MeSH
• Cell Line MeSH
• Erythrocytes immunology   MeSH
• Hemadsorption drug effects   MeSH
• Concanavalin A pharmacology   MeSH
• Chickens MeSH
• Humans MeSH
• Lymphoma MeSH
• Ovarian Neoplasms MeSH
• Receptors, Concanavalin A immunology   MeSH
• Herpesvirus 2, Gallid MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Concanavalin A MeSH
• Receptors, Concanavalin A MeSH

Breast cancer resistance protein (BCRP/ABCG2) is a member of the ATP-binding cassette transporter family that recognizes a variety of chemically unrelated compounds. Its expression has been revealed in many mammal tissues, including placenta. The purpose of this study was to describe its role in transplacental pharmacokinetics using rat placental HRP-1 cell line and dually perfused rat placenta. In HRP-1 cells, expression of Bcrp, but not P-glycoprotein, was revealed at mRNA and protein levels. Cell accumulation studies confirmed Bcrp-dependent uptake of BODIPY FL prazosin. In the placental perfusion studies, a pharmacokinetic model was applied to distinguish between passive and Bcrp-mediated transplacental passage of cimetidine as a model substrate. Bcrp was shown to act in a concentration-dependent manner and to hinder maternal-to-fetal transport of the drug. Fetal-to-maternal clearance of cimetidine was found to be 25 times higher than that in the opposite direction; this asymmetry was partly eliminated by BCRP inhibitors fumitremorgin C (2 microM) or N-(4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2-isoquinolinyl)ethyl]-phenyl)-9,10-dihydro-5-methoxy-9-oxo-4-acridine carboxamide (GF120918; 2 microM) and abolished at high cimetidine concentrations (1000 microM). When fetal perfusate was recirculated, Bcrp was found to actively remove cimetidine from the fetal compartment to the maternal compartment even against a concentration gradient and to establish a 2-fold maternal-to-fetal concentration ratio. Based on our results, we propose a two-level defensive role of Bcrp in the rat placenta in which the transporter 1) reduces passage of its substrates from mother to fetus but also 2) removes the drug already present in the fetal circulation.

• MeSH
• ATP Binding Cassette Transporter, Subfamily G, Member 2 MeSH
• ATP-Binding Cassette Transporters analysis   genetics   physiology   MeSH
• Cell Line MeSH
• Cimetidine pharmacokinetics   MeSH
• Immunohistochemistry MeSH
• Rats MeSH
• RNA, Messenger analysis   MeSH
• Metabolic Clearance Rate MeSH
• ATP Binding Cassette Transporter, Subfamily B, Member 1 physiology   MeSH
• Perfusion MeSH
• Placenta metabolism   MeSH
• Rats, Wistar MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• ATP Binding Cassette Transporter, Subfamily G, Member 2 MeSH
• ATP-Binding Cassette Transporters MeSH
• Abcg2 protein, rat MeSH Browser
• Cimetidine MeSH
• RNA, Messenger MeSH
• ATP Binding Cassette Transporter, Subfamily B, Member 1 MeSH