Pepsin Promotes Activation of Epidermal Growth Factor Receptor and Downstream Oncogenic Pathways, at Slightly Acidic and Neutral pH, in Exposed Hypopharyngeal Cells

Doukas, Panagiotis G.; Vageli, Dimitra P.; Sasaki, Clarence T.; Judson, Benjamin L.

doi:10.3390/ijms22084275

Cited by 18 publications

(17 citation statements)

References 87 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We used Cell Titer‐Glo® Luminescent Cell Viability Assay (Promega Corp., Madison, WI, USA), as previously described and in supplementary methods (Data S1), 51 to detect the effect of PARP‐1 or NF‐κB inhibition or gene silencing on acidic bile‐related cell survival, compared to acidic bile alone. Luminescence was measured by a luminometer (Gen5 software; Synergy1, BIOTEK; Gen5™ software, BioTek Instruments Inc., Winooski, VT, USA).…”

Section: Methodsmentioning

confidence: 99%

The Role of PARP‐1 and NF‐κB in Bile‐Induced DNA Damage and Oncogenic Profile in Hypopharyngeal Cells

Doukas¹,

Vageli²,

Judson³

2022

The Laryngoscope

Self Cite

View full text Add to dashboard Cite

Objectives/Hypothesis We recently documented that acidic bile, a gastroesophageal reflux content, can cause invasive hypopharyngeal squamous cell carcinoma, by inducing widespread DNA damage and promoting nuclear factor kappa B (NF‐κB)‐related oncogenic molecular events. Poly or adenosine diphosphate (ADP)‐ribose polymerase‐1 (PARP‐1), a sensitive sensor of DNA damage, may interact with NF‐κB. We hypothesized that PARP‐1 is activated in hypopharyngeal cells (HCs) with marked DNA damage caused by acidic bile, hence there is an association between PARP‐1 and NF‐κB activation or its related oncogenic profile, in this process. Study Design In vitro study. Methods We targeted PARP‐1 and NF‐κB(p65), using pharmacologic inhibitors, 1.0 μM Rucaparib (AG014699) and 10 μM BAY 11–7082 {3‐[4=methylphenyl)sulfonyl]‐(2E)‐propenenitrile}, respectively, or silencing their gene expression (siRNAs) and used immunofluorescence, luciferase, cell viability, direct enzyme‐linked immunosorbent assays, and qPCR analysis to detect the effect of targeting PARP‐1 or NF‐κB in acidic bile‐induced DNA damage, PARP‐1, p‐NF‐κB, and B‐cell lymphoma 2 (Bcl‐2) expression, as well as NF‐κB transcriptional activity, cell survival, and mRNA oncogenic phenotype in HCs. Results We showed that (i) PARP‐1 is overexpressed by acidic bile, (ii) targeting NF‐κB adequately prevents the acidic bile‐induced DNA double‐strand breaks (DSBs) by gamma H2A histone family member X (γH2AX), oxidative DNA/RNA damage, PARP‐1 overexpression, anti‐apoptotic mRNA phenotype, and cell survival, whereas (iii) targeting PARP‐1 preserves elevated DNA damage, NF‐κB activation, and anti‐apoptotic phenotype. Conclusion We document for the first time that the activation of PARP‐1 is an early event during bile reflux‐related head and neck carcinogenesis and that NF‐κB can mediate DNA damage and PARP‐1 activation. Our data encourage further investigation into how acidic bile‐induced activated NF‐κB mediates DNA damage in hypopharyngeal carcinogenesis. Level of Evidence NA Laryngoscope, 133:1146–1155, 2023

show abstract

Section: Methodsmentioning

confidence: 99%

The Role of PARP‐1 and NF‐κB in Bile‐Induced DNA Damage and Oncogenic Profile in Hypopharyngeal Cells

Doukas¹,

Vageli²,

Judson³

2022

The Laryngoscope

Self Cite

View full text Add to dashboard Cite

show abstract

“…Furthermore, Lewin et al ( 47 ) reported a close association between LPR and patients with premalignant lesions or early carcinomas of the larynx. It is worth mentioning that although there have been efforts to link the effect of other gastroesophageal refluxate contents, such as pepsin, with pre-neoplastic events in the larynx and pharynx, the conclusions have been divergent ( 48 – 54 ). A recent study indicated a possible contributory effect of slightly acidic or neutral pepsin to the inflammatory and neoplastic effects of LPR ( 54 ), but there is still no direct evidence of carcinogenesis induced by pepsin.…”

Section: Bile As a Potential Carcinogenmentioning

confidence: 99%

Bile reflux and hypopharyngeal cancer (Review)

Vageli¹,

Doukas²,

Doukas³

et al. 2021

Oncol Rep

View full text Add to dashboard Cite

Laryngopharyngeal reflux, a variant of gastroesophageal reflux disease, has been considered a risk factor in the development of hypopharyngeal cancer. Bile acids are frequently present in the gastroesophageal refluxate and their effect has been associated with inflammatory and neoplastic changes in the upper aerodigestive tract. Recent in vitro and in vivo studies have provided direct evidence of the role of acidic bile refluxate in hypopharyngeal carcinogenesis and documented the crucial role of NF-κB as a key mediator of early oncogenic molecular events in this process and also suggested a contribution of STAT3. Acidic bile can cause premalignant changes and invasive squamous cell cancer in the affected hypopharynx accompanied by DNA damage, elevated p53 expression and oncogenic mRNA and microRNA alterations, previously linked to head and neck cancer. Weakly acidic bile can also increase the risk for hypopharyngeal carcinogenesis by inducing DNA damage, exerting anti-apoptotic effects and causing precancerous lesions. The most important findings that strongly support bile reflux as an independent risk factor for hypopharyngeal cancer are presented in the current review and the underlying mechanisms are provided.

show abstract

“…IL-1β, a proinflammatory cytokine, is associated with tumor progression in lung cancer patients in multiple studies [ 53 ]. It has been recently shown that the transcriptional activation of TNF-α, IL-1β, and PTGS2 (COX-2) are among other early oncogenic molecular events that occur in the epithelium of the upper aerodigestive tract under its chronic exposure to known risk factors, such as tobacco smoke N-nitrosamines, with or without nicotine, and/or gastroesophageal refluxate, and can be prevented by the application of specific inhibitors [ 24 , 54 , 55 , 56 , 57 ]. An examination of the Cancer Genome Atlas database shows that for lung cancer, IL-1 and TNF reveal a tendency as an oncogene, as patients with higher IL-1β and TNF-α mRNA levels are linked to shorter overall survival, although this correlation does not reach statistical significance [ 57 ].…”

Section: Discussionmentioning

confidence: 99%

Sorafenib Alleviates Inflammatory Signaling of Tumor Microenvironment in Precancerous Lung Injuries

et al. 2023

Self Cite

View full text Add to dashboard Cite

According to population-based studies, lung cancer is the prominent reason for cancer-related mortality worldwide in males and is also rising in females at an alarming rate. Sorafenib (SOR), which is approved for the treatment of hepatocellular carcinoma and renal cell carcinoma, is a multitargeted protein kinase inhibitor. Additionally, SOR is the subject of interest for preclinical and clinical trials in lung cancer. This study was designed to assess in vivo the possible effects of sorafenib (SOR) in diethylnitrosamine (DEN)-induced lung carcinogenesis and examine its probable mechanisms of action. A total of 30 adult male rats were divided into three groups (1) control, (2) DEN, and (3) DEN + SOR. The chemical induction of lung carcinogenesis was performed by injection of DEN intraperitoneally at 150 mg/kg once a week for two weeks. The DEN-administered rats were co-treated with SOR of 10 mg/kg by oral gavage for 42 alternate days. Serum and lung tissue samples were analyzed to determine SRY-box transcription factor 2 (SOX-2) levels. The tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) levels were measured in lung tissue supernatants. Lung sections were analyzed for cyclooxygenase-2 (COX-2) and c-Jun N-terminal kinase (JNK) histopathologically. In addition, cyclooxygenase-2 (COX-2) and c-Jun N-terminal kinase (JNK) were analyzed by immunohistochemistry and immunofluorescence methods, respectively. SOR reduced the level of SOX-2 that maintenance of cancer stemness and tumorigenicity, and TNF-α and IL-1β levels. Histopathological analysis demonstrated widespread inflammatory cell infiltration, disorganized alveolar structure, hyperemia in the vessels, and thickened alveolar walls in DEN-induced rats. The damage was markedly reduced upon SOR treatment. Further, immunohistochemical and immunofluorescence analysis also revealed increased expression of COX-2 and JNK expression in DEN-intoxicated rats. However, SOR treatment alleviated the expression of these inflammatory markers in DEN-induced lung carcinogenesis. These findings suggested that SOR inhibits DEN-induced lung precancerous lesions through decreased inflammation with concomitant in reduced SOX-2 levels, which enables the maintenance of cancer stem cell properties.

show abstract

Pepsin Promotes Activation of Epidermal Growth Factor Receptor and Downstream Oncogenic Pathways, at Slightly Acidic and Neutral pH, in Exposed Hypopharyngeal Cells

Cited by 18 publications

References 87 publications

The Role of PARP‐1 and NF‐κB in Bile‐Induced DNA Damage and Oncogenic Profile in Hypopharyngeal Cells

The Role of PARP‐1 and NF‐κB in Bile‐Induced DNA Damage and Oncogenic Profile in Hypopharyngeal Cells

Bile reflux and hypopharyngeal cancer (Review)

Sorafenib Alleviates Inflammatory Signaling of Tumor Microenvironment in Precancerous Lung Injuries

Contact Info

Product

Resources

About