scholarly journals 17-Aminogeldanamycin Inhibits Constitutive Nuclear Factor-Kappa B (NF-κB) Activity in Patient-Derived Melanoma Cell Lines

2020 ◽  
Vol 21 (11) ◽  
pp. 3749
Author(s):  
Mariusz L. Hartman ◽  
Magdalena Rogut ◽  
Aleksandra Mielczarek-Lewandowska ◽  
Michal Wozniak ◽  
Malgorzata Czyz
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Nuclear Factor Kappa B ◽  
Time Lapse ◽  
Genetic Alterations ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nuclear Factor ◽  
Nuclear Factor Kappa ◽  
Promising Therapeutic Approach ◽  
Melanoma Cell Lines

Melanoma remains incurable skin cancer, and targeting heat shock protein 90 (HSP90) is a promising therapeutic approach. In this study, we investigate the effect of 17-aminogeldanamycin, a potent HSP90 inhibitor, on nuclear factor-kappa B (NF-κB) activity in BRAFV600E and NRASQ61R patient-derived melanoma cell lines. We performed time-lapse microscopy and flow cytometry to monitor changes in cell confluence and viability. The NF-κB activity was determined by immunodetection of phospho-p65 and assessment of expression of NF-κB-dependent genes by quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and enzyme-linked immunosorbent assay (ELISA). Constitutive activity of p65/NF-κB was evident in all melanoma cell lines. Differences in its level might be associated with genetic alterations in CHUK, IL1B, MAP3K14, NFKBIE, RIPK1, and TLR4, while differences in transcript levels of NF-κB-inducible genes revealed by PCR array might result from the contribution of other regulatory mechanisms. 17-Aminogeldanamycin markedly diminished the level of phospho-p65, but the total p65 protein level was unaltered, indicating that 17-aminogeldanamycin inhibited activation of p65/NF-κB. This conclusion was supported by significantly reduced expression of selected NF-κB-dependent genes: cyclin D1 (CCND1), C-X-C motif chemokine ligand 8 (CXCL8), and vascular endothelial growth factor (VEGF), as shown at transcript and protein levels, as well as secretion of IL-8 and VEGF. Our study indicates that 17-aminogeldanamycin can be used for efficient inhibition of NF-κB activity and the simultaneous diminution of IL-8 and VEGF levels in the extracellular milieu of melanoma.

scholarly journals Circulating Nuclear Factor-Kappa B Mediates Cancer-Associated Inflammation in Human Breast and Colon Cancer

2020 ◽  
Author(s):  
Hafize Uzun ◽  
Berrin Papila Kundaktepe ◽  
Volkan Sozer ◽  
Pinar Kocael ◽  
Sinem Durmus ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Colon Cancer ◽  
Nuclear Factor Kappa B ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nuclear Factor ◽  
Healthy Controls ◽  
Nuclear Factor Kappa ◽  
Tnf Α ◽  
Study Results ◽  
Serum Crp

Background: Inflammation is recognized as a hallmark feature of cancer development and progression. The aim of our study was to investigate the significance of serum nuclear factor kappa-B (NF-кB) levels as a circulating marker in the monitor of inflammation in breast and colon cancer; to show the relationship between NF-кB with inflammatory parameters as tumor necros faktor-α (TNF-α), soluble TNF-related apoptosis inducing ligand (sTRAIL), interleukin-6 (IL-6), pentraxin-3 (PTX-3), procalcitonin (PCT), and C-reactive protein (CRP) levels. Methods: Serum NF-кB, TNF-α, sTRAIL, IL-6, PTX-3, PCT, and serum CRP levels were measured using enzyme linked immunosorbent assay (ELISA) in 40 patients with breast cancer, 40 patients with colon cancer and 30 healthy controls. Results: The serum NF-кB, TNF-α, IL-6, PTX-3, PCT and serum CRP concentration was significantly higher and the serum sTRAIL concentration was significantly lower in patients with breast and colon cancer than in those with an healthy controls. NF-кB were positive correlated with CRP and negative corelated with sTRAIL. Conclusions: These results suggest that increased NF-кB may decrease the clinical efficacy of sTRAIL in solid tumour cells. There is relationship between inflammation and carcinogenesis so that the development of cancer occurs with chronic inflammation in breast and colon. The study results have shown that colon and breast cancer patients have increased systemic inflammation, as measured by increased circulating cytokines, and acute phase proteins, or by abnormalities in circulating cells. NF-кB may combine with other markers of the systemic inflammatory response in prognostic scores in cancer. In addition to surgical resection of the tumor, conventional radio- and chemotherapy for cancer treatment, the use of sTRAIL or other agonists for cancer therapy appeared a new potential therapy.  

scholarly journals The YUMM lines: a series of congenic mouse melanoma cell lines with defined genetic alterations

2016 ◽  
Vol 29 (5) ◽  
pp. 590-597 ◽  
Author(s):  
Katrina Meeth ◽  
Jake Xiao Wang ◽  
Goran Micevic ◽  
William Damsky ◽  
Marcus W. Bosenberg
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Genetic Alterations ◽  
Congenic Mouse ◽  
Mouse Melanoma ◽  
Mouse Melanoma Cell ◽  
Melanoma Cell Lines

scholarly journals Biotransformed Soybean Extract (BSE) Inhibits Melanoma Cell Growth and Viability In Vitro: Involvement of Nuclear Factor-Kappa B Signaling

PLoS ONE ◽  
2014 ◽  
Vol 9 (7) ◽  
pp. e103248 ◽  
Author(s):  
Fernanda Maria Pinto Vilela ◽  
Deeba N. Syed ◽  
Jean Christopher Chamcheu ◽  
Laura A. Calvo-Castro ◽  
Vanessa Silveira Fortes ◽  
...  
Keyword(s):  
Cell Growth ◽  
Melanoma Cell ◽  
Nuclear Factor Kappa B ◽  
Nuclear Factor ◽  
Nuclear Factor Kappa ◽  
Soybean Extract ◽  
Cell Growth And Viability

scholarly journals The Relationship between Serum and Gene Expression Levels of RANK, RANKL and Osteoprotegerin Inflammatory Pathway with Unstable Angina: A Case-control Study

2021 ◽  
Author(s):  
Alireza Farrokhian ◽  
Mahtab Miraftab ◽  
Minoo Chenari ◽  
Hossein Akbari ◽  
Hassan Nikoueinejad ◽  
...  
Keyword(s):  
Gene Expression ◽  
Unstable Angina ◽  
Nuclear Factor Kappa B ◽  
Mononuclear Cells ◽  
Serum Levels ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nuclear Factor ◽  
Inflammatory Pathway ◽  
Nuclear Factor Kappa ◽  
Receptor Activator

Osteoprotegerin (OPG), receptor activator of nuclear factor-kappa B (RANK) and receptor activator of nuclear factor-kappa B ligand (RANKL), the members of the tumor necrosis factor (TNF) family, have multiple effects on bone metabolism, endocrine functions and, as an inflammatory pathway, in the immune system. This study tried to determine the association of the OPG/RANKL/RANK axis with the severity of unstable angina (UA) as an inflammatory condition. Our study involved 50 patients with UA and 50 healthy people. Serum and peripheral blood mononuclear cells were isolated from all participants. Serum levels and gene expression of OPG, RANKL, and RANK in mononuclear cells were measured by enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR), respectively. For each patient with UA, the thrombolysis in myocardial infarction (TIMI) and the global registry of acute coronary events (GRACE) scores were determined to evaluate the severity of the disease. Then we analyzed the relation of OPG, RANKL, and RANK levels with TIMI and GRACE scores in patients with UA. Discriminate analysis was used to predict the combinational models of such factors on the prediction of UA. Serum levels of OPG and RANKL (p<0.001) and gene expression of RANKL (p<0.001) were significantly more in patients than those in healthy ones. No relation was seen between the OPG/RANKL/RANK axis and the severity of UA according to TIMI and GRACE scores. Our study shows that serum level, as well as gene expression of OPG/RANKL/RANK axis neither, predicts the occurrence of UA nor shows any relationship with its severity.  

scholarly journals The Long Noncoding RNA ANRIL Promotes Cell Apoptosis in Lipopolysaccharide-Induced Acute Kidney Injury Mediated by the TLR4/Nuclear Factor-Kappa B Pathway

2020 ◽  
Vol 45 (2) ◽  
pp. 209-221 ◽  
Author(s):  
Ye Zhu ◽  
Sheng-Wei Wei ◽  
Ao Ding ◽  
Wei-Ping Zhu ◽  
Mei-Fang Mai ◽  
...  
Keyword(s):  
Noncoding Rna ◽  
Nuclear Factor Kappa B ◽  
Quantitative Polymerase Chain Reaction ◽  
Kidney Tissue ◽  
Inflammatory Factors ◽  
Luciferase Reporter ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Nuclear Factor ◽  
Nuclear Factor Kappa

Background/Aims: The purpose of this study is to analyze the expression and biological function of lncRNA ANRIL, microRNA-199a, TLR4, and nuclear factor-kappa B (NF-κB) in acute renal injury (AKI) induced by lipopolysaccharide (LPS). Methods: The levels of ANRIL and microRNA-199a in mouse cells and kidneys were detected by quantitative-polymerase chain reaction. Western blot analysis was used for the NF-κB pathway protein. MTT assay was used for cell viability. Enzyme-linked immunosorbent assay was used for the secretion of inflammatory factors in mouse kidney tissue. Apoptosis was measured by flow cytometry and Western blotting. The potential binding region between ANRIL and miR-199a was verified by luciferase reporter assay. Results: The upregulation of ANRIL can reduce the expression of microRNA-199a and increases the number of apoptotic cells. The expression levels of ANRIL in LPS-induced AKI mice and LPS-treated HK2 cells were upregulated compared with the control group. Overexpression of ANRIL increased apoptosis and promoted TLR4 (Toll-like receptor 4), NF-κB phosphorylation, and downstream transcription factor production. Conclusion: ANRIL/NF-κB pathway in LPS-induced apoptosis provided theoretical guidance for ANRIL in the treatment of AKI.

scholarly journals 2441 INTERLEUKIN-1B AND CYCLOOXYGENASE-2 PROINFLAMMATION ANALYSIS AND IN SILICO DOCKING NUCLEAR FACTOR KAPPA B ON ENDOMETRIOSIS CELL CULTURE GIVEN HEPTYL GALLATE AND OCTYL GALLATE TREATMENT

2019 ◽  
pp. 503-506
Author(s):  
ARLENI BUSTAMI ◽  
FAJAR SULISTYA UTAMI ◽  
RAHMI BUDIARTI ◽  
HERI WIBOWO
Keyword(s):  
Cell Culture ◽  
Gallic Acid ◽  
In Silico ◽  
Nuclear Factor Kappa B ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nuclear Factor ◽  
In Silico Docking ◽  
Nuclear Factor Kappa ◽  
Proinflammatory Factor ◽  
Cox 2

Objective: The aim of this study is to analyze the effect of octyl gallate and heptyl gallate toward the regulation of interleukin-1β and cyclooxygenase (COX)-2 proinflammatory factor on endometriosis cell culture and analyze its activity toward nuclear factor kappa B (NFkB) target protein through in silico docking technique. Methods: In vitro study was performed on endometriosis cells cultured treated with two dosages each of heptyl and octyl gallate (51.2 μg/mL and 102.4 μg/mL) for 48 h, then followed by 10 ng/mL lipopolysaccharides (LPS) induction for 24 h. The positive control group was treated by LPS induced and the negative control was treated without LPS. Inflammation regulation was evaluated with enzyme-linked immunosorbent assay technique and in silico docking analyzed using bioinformatics technique. Results: Molecular docking analysis with gallic acid and their derivatives showed that more stable affinity and stronger binding found on octyl gallate than heptyl gallate and gallic acid at the active site of NFkB. Conclusions: Based on this study results, octyl gallate and heptyl gallate were proven to be able to reduce COX-2 proinflammatory factor through NFkB pathway as an inflammatory regulator; thus, it has the potential to be developed as a therapy for endometriosis.

Sequentially-accumulated genetic alterations at chromosome 9p21 play a role in melanoma progression

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 8522-8522
Author(s):  
G. Palmieri ◽  
C. Rozzo ◽  
A. Manca ◽  
A. Cossu ◽  
A. Lissia ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Genetic Alterations ◽  
Allelic Loss ◽  
Chromosome 9P21 ◽  
Cdkn2a Gene ◽  
Melanoma Progression ◽  
Dysplastic Nevi ◽  
Tumor Dissemination ◽  
Melanoma Cell Lines

8522 Background: Cytogenetic and molecular studies indicated the chromosome 9p21 and its CDKN locus, with the p16/CDKN2A tumor suppressor genes, as the genomic regions involved into the pathogenesis of malignant melanoma (MM). To further elucidate the role of such regions in melanoma, we evaluated the 9p21-linked molecular alterations during the different phases of melanocytic tumorigenesis. Methods: Paraffin-embedded tissue sections from common nevi, dysplastic nevi, and melanomas, including primary and metastatic MM lesion, as well as melanoma cell lines were evaluated by both fluorescence in situ hybridization (FISH) using probes spanning the entire 9p21 region and immunohistochemistry (IHC) for p16/CDKN2A expression. Results: Allelic deletion at the CDKN locus and p16/CDKN2A gene silencing were both observed at increased rates moving from nevi to early or advanced primary melanomas and to correspondent metastases. Dysplastic nevi, primary and secondary melanomas were instead found to be heterozygously deleted at one or more loci within the 9p21 chromosome in a quite similar fraction (55–62%) of cases. Inactivation of the p16/CDKN2A gene was found at rates higher in melanoma cell lines (67%) than in vivo melanomas (62% secondary and 12.5% primary melanomas), with a normal p16/CDKN2A IHC staining in all analyzed nevi. Conclusions: Our findings indicate a model of sequential accumulation of genetic alterations from normal melanocytes to metastatic melanoma, with a) 9p21 allelic loss playing a role in melanocytic transformation and tumor initiation; b) deletions at CDKN locus and p16/ CDKN2A gene inactivation participating to tumor dissemination. Presence of such alterations could be further evaluated as markers of the different phases of melanoma progression. No significant financial relationships to disclose.

scholarly journals Proteasome 26S subunit, non-ATPases 1 (PSMD1) and 3 (PSMD3), play an oncogenic role in chronic myeloid leukemia by stabilizing nuclear factor-kappa B

Oncogene ◽  
2021 ◽  
Vol 40 (15) ◽  
pp. 2697-2710
Author(s):  
Alfonso E. Bencomo-Alvarez ◽  
Andres J. Rubio ◽  
Idaly M. Olivas ◽  
Mayra A. Gonzalez ◽  
Rebecca Ellwood ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Nuclear Factor Kappa B ◽  
Luciferase Reporter ◽  
Nuclear Factor ◽  
Nuclear Factor Kappa ◽  
Tki Resistance ◽  
Reporter Assays ◽  
Proteasome 26S

AbstractTyrosine kinase inhibitors (TKIs) targeting BCR-ABL1 have revolutionized therapy for chronic myeloid leukemia (CML), paving the way for clinical development in other diseases. Despite success, targeting leukemic stem cells and overcoming drug resistance remain challenges for curative cancer therapy. To identify drivers of kinase-independent TKI resistance in CML, we performed genome-wide expression analyses on TKI-resistant versus sensitive CML cell lines, revealing a nuclear factor-kappa B (NF-κB) expression signature. Nucleocytoplasmic fractionation and luciferase reporter assays confirmed increased NF-κB activity in the nucleus of TKI-resistant versus sensitive CML cell lines and CD34+ patient samples. Two genes that were upregulated in TKI-resistant CML cells were proteasome 26S subunit, non-ATPases 1 (PSMD1) and 3 (PSMD3), both members of the 19S regulatory complex in the 26S proteasome. PSMD1 and PSMD3 were also identified as survival-critical genes in a published small hairpin RNA library screen of TKI resistance. We observed markedly higher levels of PSMD1 and PSMD3 mRNA in CML patients who had progressed to the blast phase compared with the chronic phase of the disease. Knockdown of PSMD1 or PSMD3 protein correlated with reduced survival and increased apoptosis in CML cells, but not in normal cord blood CD34+ progenitors. Luciferase reporter assays and immunoblot analyses demonstrated that PSMD1 and PSMD3 promote NF-κB protein expression in CML, and that signal transducer and activator of transcription 3 (STAT3) further activates NF-κB in scenarios of TKI resistance. Our data identify NF-κB as a transcriptional driver in TKI resistance, and implicate PSMD1 and PSMD3 as plausible therapeutic targets worthy of future investigation in CML and possibly other malignancies.

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