Neuropilin-1 expression in adenocarcinoma and squamous cell carcinoma of the lung is differentially regulated by hypoxia

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 17152-17152
Author(s):  
G. P. Pidgeon ◽  
M. P. Barr ◽  
M. C. Cathcart ◽  
S. Gray ◽  
K. J. O’Byrne
Keyword(s):  
Lung Cancer ◽  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
A549 Cells ◽  
Lung Tumours ◽  
Western Analysis ◽  
Neuropilin 1

17152 Background: Neuropilin-1 (NRP-1) is an isoform-specific receptor for VEGF165 and semaphorin3A, initially discovered on migrating neurons. NRP-1 expression has been reported on a number of tumour cell lines in the absence of the other VEGF receptors, where it mediates survival signals. In this study we examined the regulation of NRP-1 by hypoxia, its effect on survival in a panel of lung cancer cell lines and its potential as a biomarker in retrospective human lung tumours. Methods: A549, SK-MES1, H460 and H647 cells were grown in serum depleted media (0.5%) in normoxic or hypoxic (0.1% O2) conditions and screened for NRP-1 expression by western and immunocytochemistry analysis. Cell survival and apoptosis was determined using BrdU and Annexin-V/PI staining respectively following treatment with an antibody to the extracellular NRP-1 domain. A panel of 100 retrospective resected lung tumours and matched normal samples were stained for NRP-1 expression by immunhistochemistry. Results: A549, SKMES-1 and H647 cell lines all expressed NRP-1 and displayed reduced survival following treatment with NRP-1 antibody (1ug/ml) compared to controls (A549 46%, SKMES-1 61%, H647 53%). H460 did not express NRP-1 and no survival inhibition was seen in the cell line (104%). Reduced survival was accompanied by increased apoptosis in all NRP-1 positive cell lines. Hypoxia strongly increased NRP-1 expression in the A549 adenocarcinoma (AC) cell line, while NRP-1 was decreased in SKMES-1 squamous cell carcinoma (SCC) following hypoxia. Neutralisation of NRP-1 had a greater effect in A549 cells under hypoxia (37%), with a lesser effect in SKMES-1 cells (82%). Western analysis of matched frozen normal and lung cancer biopsies showed NRP-1 overexpression in AC and decreased expression in SCC relative to normal. High NRP-1 expression was confirmed in AC and large cell carcinoma by immunohistochemistry, relative to normal. However, SCCs had a lower level of NRP-1 staining, supporting the results by western analysis and following hypoxia in vitro. Conclusions: These results implicate NRP-1 as an important survival pathway in lung cancer. Hypoxia differentially regulated NRP-1 mediated survival implicating this pathway as a potential therapeutic strategy in AC. No significant financial relationships to disclose.

scholarly journals Spontaneous changes in intermediate filament protein expression patterns in lung cancer cell lines

1988 ◽  
Vol 91 (1) ◽  
pp. 91-108
Author(s):  
J.L. Broers ◽  
M.K. Rot ◽  
T. Oostendorp ◽  
G. Bepler ◽  
L. De Leij ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cell Carcinoma ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
Expression Patterns ◽  
Small Cell ◽  
Intermediate Filament Protein ◽  
Lung Cancer Cell ◽  
Filament Protein

The usefulness of cell lines in the study and prediction of the clinical behaviour of lung cancer is still a matter of debate. However, lung tumour cell cultures have been of value in investigations concerning molecular and cell biological aspects of these neoplasms. Especially in the examination of characteristics specific for the main types of differentiation (squamous cell carcinoma, adenocarcinoma, small cell carcinoma), in vitro studies have been most important. Twenty eight lung cancer cell lines were cultured for up to four years, and were examined at regular intervals for their intermediate filament protein (IFP) expression patterns using a panel of cytokeratin (CK) and neurofilament (NF) antibodies. These studies showed that the classic type of small cell lung cancer (SCLC) cell lines contain CKs 8, 18, and occasionally CK 19, while the variant-type SCLC cell lines generally express no CKs but can contain NFs. Non-SCLC cell lines, such as squamous cell carcinoma and adenocarcinoma cell lines, contain CKs 7 (in most cases), 8, 18 and 19. In one variant SCLC cell line and in one adenocarcinoma cell line CKs 4, 10 and 13, characteristic of squamous cell differentiation, were found. Although most cell lines have remained stable with respect to growth characteristics and IFP expression patterns, five lung cancer cultures exhibited a transition from one cell type to another, paralleled by changes in IFP expression. Progressions from classic to variant SCLC cell lines have been observed, next to conversions from variant SCLC to cell lines re-expressing cytokeratins. In some cases this resulted in a coexpression of CKs and NFs within a cell line and even within individual tumour cells. These results strongly support the earlier finding that CK expression in SCLC cell lines is a reliable marker for the classic type of differentiation, while the absence of CKs and the presence of NFs marks the variant type of differentiation. Our results are discussed in view of previous histological findings.

scholarly journals Remedial Effects and Molecular Docking Studies of Nevadensin: ‎Antiglucosidase, Anti-cholinesterase, and Anti-Human Oral Squamous ‎Cell Carcinoma Potentials

2021 ◽  
Author(s):  
Xu Lan ◽  
Li Liu ◽  
Guangyu Wu
Keyword(s):  
Squamous Cell Carcinoma ◽  
Molecular Docking ◽  
Oral Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
Antioxidant Activities ◽  
Biological Activities ◽  
Radical Scavenging

IntroductionNevadensin has a variety of pharmacological effects, including effects of anti-mycobacterium ‎tuberculosis, antitussive, anti-hypertensive, anti-inflammatory, and free radical-scavenging ‎activities. In this study, we investigated for their anticholinergics, antidiabetic, and anti-‎human oral squamous cell carcinoma potentials for nevadensin. ‎Material and methodsThe antioxidant activities of nevadensin were elucidated by using various bioanalytical assays. ‎On the other hand, IC50 values were calculated for acetylcholine esterase, α-glucosidase ‎inhibition effects of nevadensin. For determining of anti-human oral squamous cell carcinoma ‎properties of nevadensin, MTT assay was used on HUVEC, HSC-3, HSC-4, and Ca9-22 cell ‎lines. The molecular docking method used to compare the biological activities of the ‎nevadensin molecule against enzymes was used. Afterwards, the ADME/T analysis was ‎performed to investigate the drug availability of the nevadensin molecule and the obtained ‎parameters from ADME/T analysis were examined.‎ResultsThe cell viability of nevadensin was very low against human oral squamous cell carcinoma cell ‎lines without any cytotoxicity on the human normal (HUVEC) cell line. The IC50 of the ‎nevadensin against HSC-3, HSC-4, and Ca9-22 were 316, 273, and 399 µg/mL, respectively. ‎Thereby, the best cytotoxicity results and anti-human oral squamous cell carcinoma potentials ‎of our nevadensin was observed in the case of the HSC-4 cell line. ‎ConclusionsMaybe the anti-human lung carcinoma properties of nevadensin are related to their antioxidant ‎effects. ‎

scholarly journals Tongue Cancer and Epigenetic Factors: An in-vitro Study on 298 Micro-RNAS

2012 ◽  
Vol 10 (3) ◽  
pp. 447-454
Author(s):  
A. Guida ◽  
G. Pannone ◽  
A. Lucchese ◽  
R. Serpico ◽  
D. Pasquali ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
Tongue Cancer ◽  
In Vitro Study ◽  
Early Results ◽  
Micro Rnas

Oral Squamous Cell Carcinoma (OSCC) is the sixth most frequent malignant tumour. There is some evidence that tongue cancer has a higher local failure rate and poorer prognosis than other anatomical sites in the oral cavity. We used tongue squamous cell carcinoma cell lines harbouring mutated p53/p16 as tongue cancer models to study the influences exerted by p53 and p16 genes on the expression of micro RNAs (miRNAs). The study was performed on microarray chips harbouring 298 miRNA sequences. OSCC cell lines used in this study were SCC-4, SCC-15 and SCC-25, all three carrying mutated/hypermethylated p53/p16. The expression values normalized to healthy control of 298 miRNAs were obtained for each cell line. MiRNA 196b was found hyperexpressed in the three cell lines. MiRNAs 19b-1, 21, 27a, 30d, 134, 339, 379 and 465 were found altered in two out of three cell lines. miRNAs found altered in one cell line out of three were: 7b, 23a, 25, 30c, 30e-3p, 107,125b, 124a, 214, 216, 325 and 384. A literature review for each miRNA found significant was undertaken. Some miRNAs have a well-known role in oral cancer, some have been put in correlation with other cancers/diseases, others are found significant for the first time. These early results in tongue cancer cell lines harbouring mutation of p16/p53 need further analyses to understand whether this variation of miRNA levels are directly influenced by the malfunction of these proteins or if, vice-versa, altered miRNA levels influence the function of p16 and p53.

scholarly journals Targeting PKCι-PAK1 signaling pathways in EGFR and KRAS mutant adenocarcinoma and lung squamous cell carcinoma

2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Masaoki Ito ◽  
Carles Codony-Servat ◽  
Jordi Codony-Servat ◽  
David Lligé ◽  
Imane Chaib ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Squamous Cell Carcinoma ◽  
Cell Viability ◽  
Cell Carcinoma ◽  
Signaling Pathways ◽  
Cell Lines ◽  
Squamous Cell ◽  
Drug Combination ◽  
Colony Formation ◽  
Tumor Volume

Abstract Introduction p21-activated kinase 1 (PAK1) stimulates growth and metastasis in non-small cell lung cancer (NSCLC). Protein kinase C iota (PKCι) is an enzyme highly expressed in NSCLC, regulating PAK1 signaling. In the present study we explored whether the PKCι-PAK1 signaling pathway approach can be an efficient target in different types of NSCLC cell and mouse models. Methods The effect of IPA-3 (PAK1 inhibitor) plus auranofin (PKCι inhibitor) combination was evaluated by cell viability assay, colony formation and western blotting assay, using three types of NSCLC cell lines: EGFR or KRAS mutant adenocarcinoma and squamous cell carcinoma with PAK1 amplification. In addition, for clinical availability, screening for new PAK1 inhibitors was carried out and the compound OTSSP167 was evaluated in combination with auranofin in cell and mice models. Results The combination of IPA-3 or OTSSP167 plus auranofin showed high synergism for inhibiting cell viability and colony formation in three cell lines. Mechanistic characterization revealed that this drug combination abrogated expression and activation of membrane receptors and downstream signaling proteins crucial in lung cancer: EGFR, MET, PAK1, PKCι, ERK1/2, AKT, YAP1 and mTOR. A nude mouse xenograft assay demonstrated that this drug combination strongly suppressed tumor volume compared with single drug treatment. Conclusions Combination of IPA-3 or OTSSP167 and auranofin was highly synergistic in EGFR or KRAS mutant adenocarcinoma and squamous cell carcinoma cell lines and decreased tumor volume in mice models. It is of interest to further test the targeting of PKCι-PAK1 signaling pathways in EGFR mutant, KRAS mutant and squamous NSCLC patients.

scholarly journals Cancer cell line microarray as a novel screening method for identification of radioresistance biomarkers in head and neck squamous cell carcinoma

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Johannes Routila ◽  
Karri Suvila ◽  
Reidar Grénman ◽  
Ilmo Leivo ◽  
Jukka Westermarck ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Stem Cell ◽  
Cell Carcinoma ◽  
Head And Neck ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
Stem Cell Marker ◽  
Cell Marker ◽  
Hnscc Cell

Abstract Background Currently, no clinically useful biomarkers for radioresistance are available in head and neck squamous cell carcinoma (HNSCC). This study assesses the usefulness of Cell Line Microarray (CMA) method to enhance immunohistochemical screening of potential immunohistochemical biomarkers for radioresistance in HNSCC cell lines. Methods Twenty-nine HNSCC cell lines were cultured, cell pellets formalin-fixed, paraffin-embedded, and arrayed. Radioresistance features of the cell lines were combined to immunohistochemical stains for p53, NDFIP1, EGFR, stem cell marker Oct4, and PP2A inhibitor CIP2A. Results Expression of p53, EGFR or CIP2A did not indicate intrinsic radioresistance in vitro. Stem cell marker Oct4 nuclear positivity and NDFIP1 nuclear positivity was correlated with increased intrinsic radioresistance. Conclusion The usefulness of CMA in analysis of HNSCC cell lines and discovery of biomarkers is demonstrated. CMA is very well adapted to both testing of antibodies in a large panel of cell lines as well as correlating staining results with other cell line characteristics. In addition, CMA-based antibody screening proved an efficient and relatively simple method to identify potential radioresistance biomarkers in HNSCC.

scholarly journals Pharmacological Inhibition of PP2A Overcomes Nab-Paclitaxel Resistance by Downregulating MCL1 in Esophageal Squamous Cell Carcinoma (ESCC)

Cancers ◽  
2021 ◽  
Vol 13 (19) ◽  
pp. 4766
Author(s):  
Qi Song ◽  
Herui Wang ◽  
Dongxian Jiang ◽  
Chen Xu ◽  
Jing Cui ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Esophageal Squamous Cell Carcinoma ◽  
Tumor Growth ◽  
Cell Carcinoma ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
Resistant Cell ◽  
Parental Cell Line ◽  
Pharmacological Inhibition

Paclitaxel-based chemotherapy is a treatment option for advanced esophageal squamous cell carcinoma (ESCC). However, the development of chemoresistance leads to treatment failure, and the underlying mechanism remains elusive. We investigated the mechanisms of nanoparticle albumin-bound paclitaxel (nab-PTX) resistance by establishing three nab-PTX resistant ESCC cell lines. Proteomics analysis revealed higher oxidative phosphorylation (OXPHOS) in resistant cell line DR150 than in its parental cell line KYSE150, which is likely caused by stabilized anti-apoptotic protein MCL1. Additionally, we discovered the elevated activity of protein phosphatase 2A (PP2A), the phosphatase that dephosphorylates and stabilizes MCL1, in nab-PTX resistant cell lines. Pharmacological inhibition of PP2A with small molecule compound LB-100 decreased MCL1 protein level, caused more apoptosis in nab-PTX resistant ESCC cell lines than in the parental cells in vitro, and significantly inhibited the tumor growth of nab-PTX resistant xenografts in vivo. Moreover, LB-100 pretreatment partially restored nab-PTX sensitivity in the resistant cell lines and synergistically inhibited the tumor growth of nab-PTX resistant xenografts with nab-PTX. In summary, our study identifies a novel mechanism whereby elevated PP2A activity stabilizes MCL1 protein, increases OXPHOS, and confers nab-PTX resistance, suggesting that targeting PP2A is a potential strategy for reversing nab-PTX resistance in patients with advanced ESCC.

scholarly journals Two alternative cell line models for the study of multiorganic metastasis and immunotherapy in Lung Squamous Cell Carcinoma

2021 ◽  
Author(s):  
Karmele Valencia ◽  
Cristina Sainz ◽  
Cristina Bértolo ◽  
Gabriel de Biurrun ◽  
Jackeline Agorreta ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
Lung Squamous Cell Carcinoma ◽  
Metastatic Potential ◽  
Human Squamous Cell Carcinoma ◽  
Whole Exome

There is a paucity of adequate mouse models and cell lines available to study lung squamous cell carcinoma (LUSC). We have generated and characterized two models of phenotypically different transplantable LUSC cell lines (UN-SCC679 and UN-SCC680) derived from an N-nitroso-tris-chloroethylurea (NTCU) chemically-induced mouse model in A/J mice. Furthermore, we genetically characterized and compared both LUSC cell lines by performing whole exome and RNA sequencing. These experiments revealed similar genetic and transcriptomic patterns that may correspond to the classical LUSC human subtype. In addition, we compared the immune landscape generated by both tumor cells lines in vivo and assessed their response to immune checkpoint inhibition. The differences between the two cell lines are a good model for the remarkable heterogeneity of human squamous cell carcinoma. Study of the metastatic potential of these models revealed that both cell lines represent the human LUSC organotropism to the brain, bones, liver and adrenal glands. In summary, we have generated a very valuable cell line tools for LUSC research that recapitulates the complexity of the human disease.

scholarly journals Identification and Characterization of Cancer Stem Cells from Head and Neck Squamous Cell Carcinoma Cell Lines

2015 ◽  
Vol 36 (2) ◽  
pp. 784-798 ◽  
Author(s):  
Valentina Pozzi ◽  
Davide Sartini ◽  
Romina Rocchetti ◽  
Andrea Santarelli ◽  
Corrado Rubini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Squamous Cell Carcinoma ◽  
Cancer Stem Cells ◽  
Cell Carcinoma ◽  
Head And Neck ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell

Background/Aims: Head and neck squamous cell carcinoma (HNSCC) ranks sixth worldwide for tumor-related mortality. A subpopulation of tumor cells, termed cancer stem cells (CSCs), has the ability to support cancer growth. Therefore, profiling CSC-enriched populations could be a reliable tool to study cancer biology. Methods: We performed phenotypic characterization of 7 HNSCC cell lines and evaluated the presence of CSCs. CSCs from Hep-2 cell line and HNSCC primary cultures were enriched through sphere formation and sphere-forming cells have been characterized both in vitro and in vivo. In addition, we investigated the expression levels of Nicotinamide N-methyltransferase (NNMT), an enzyme overexpressed in several malignancies. Results: CSC markers were markedly expressed in Hep-2 cell line, which was found to be highly tumorigenic. CSC-enriched populations displayed increased expression of CSC markers and a strong capability to form tumors in vivo. We also found an overexpression of CSC markers in tumor formed by CSC-enriched populations. Interestingly, NNMT levels were significantly higher in CSC-enriched populations compared with parental cells. Conclusion: Our study provides an useful procedure for CSC identification and enrichment in HNSCC. Moreover, results obtained seem to suggest that CSCs may represent a promising target for an anticancer therapy.

scholarly journals PD-L1 and MRN synergy in platinum-based chemoresistance of head and neck squamous cell carcinoma

2019 ◽  
Vol 122 (5) ◽  
pp. 640-647 ◽  
Author(s):  
Bin Shen ◽  
Dongyan Huang ◽  
Andrew J. Ramsey ◽  
Kevin Ig-Izevbekhai ◽  
Kevin Zhang ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Head And Neck ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
Molecular Mechanisms ◽  
Cisplatin Resistance ◽  
Neck Squamous Cell Carcinoma ◽  
Hnscc Cell

Abstract Background We have been investigating the molecular mechanisms of cisplatin-induced chemoresistance in head and neck squamous cell carcinoma (HNSCC). Based on our previous findings, the present study investigates how the Mre11, Rad50, and NBS1 (MRN) DNA repair complex interacts at the molecular level with the programmed cell death ligand 1 (PD-L1) in cisplatin-induced chemoresistance. Methods Human HNSCC cell lines were used to determine the role played by PD-L1 in cisplatin resistance. Initial experiments investigated PD-L1 expression levels in cells exposed to cisplatin and whether PD-L1 interacts directly with the MRN complex. Finally, in vitro studies and in vivo experiments on BALB/c nu/nu mice were performed to determine whether interference of PD-L1 or NBS1 synthesis modulated cisplatin resistance. Results Exposure to cisplatin resulted in PD-L1 being upregulated in the chemoresistant but not the chemosensitive cell line. Subsequent co-immunoprecipitation studies demonstrated that PD-L1 associates with NBS1. In addition, we found that the knockdown of either PD-L1 or NBS1 re-sensitised the chemoresistant cell line to cisplatin. Finally, but perhaps most importantly, synergy was observed when both PD-L1 and NBS1 were knocked down making the formerly chemoresistant strain highly cisplatin sensitive. Conclusions PD-L1 plays a pivotal role in cisplatin resistance in chemoresistant human HNSCC cell lines.

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