Experimental carcinogenesis on the oropharyngeal mucosa of rats with hydrochloric acid, sodium nitrate and pepsin

Negro, André Del; Araújo, Marina Raquel; Tincani, Álfio José; Meirelles, Luciana Rodrigues de; Martins, Alessandro F.; Andreollo, Nelson Adami

doi:10.1590/s0102-86502008000400007

Cited by 10 publications

(9 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Adams favors the preneoplastic role of combined pepsin and acid [39] , whereas Bock and Johnston defend the effect of nonacidic pepsin [40] , [41] . On the other hand, Del Negro in an in vivo rat model describes no evidence of neoplastic activity with pepsin and acid [42] .…”

Section: Discussionmentioning

confidence: 93%

miR-21, miR-155, miR-192, and miR-375 Deregulations Related to NF-kappaB Activation in Gastroduodenal Fluid–Induced Early Preneoplastic Lesions of Laryngeal Mucosa In Vivo

Sasaki¹,

Vageli²

2016

Neoplasia

View full text Add to dashboard Cite

Gastroduodenal refluxate found in the upper aerodigestive tract is not clinically uncommon. We recently demonstrated the neoplastic potential of gastroduodenal fluids (GDF) on hypopharyngeal mucosa, via NF-κB, using in vitro and in vivo models. Here we will explore the in vivo effect of GDF on laryngeal mucosa (LM) to induce early preneoplastic lesions related to NF-κB activation, along with deregulation of specific microRNA (miRNA) markers previously linked to laryngeal cancer. We used histological, immunohistochemical, automated quantitative analysis and quantitative polymerase chain reaction to examine LM from 35 C57Bl/6J mice previously treated with topical GDF against corresponding controls (4 experimental and 3 control groups; 5 mice/group). Our analysis showed that GDF produced early preneoplastic lesions in treated LM related to NF-κB activation. LM treated by acid and bile combination demonstrated significantly higher expression of the analyzed cell proliferation markers (Ki67, CK14, ∆Np63), oncogenic p-STAT3, and changes of cell adhesion molecules (E-cadherin, ϐ-catenin) versus untreated LM or LM exposed to acid alone (P < .0005). Furthermore, acidic bile but not neutral bile appeared to accelerate the expression of “oncomirs” miR-21, miR-155, and miR-192 (acidic bile versus neutral bile, P < .0001), while reducing tumor suppressor miR-375 (acidic bile versus neutral bile, P = .0137), previously linked to NF-κB and laryngeal cancer. Finally, acidic bile induced reduction of miR-34a, miR-375, and miR-451a, exhibiting an inverse correlation with NF-κB activation. SIGNIFICANCE: Bile in combination with acid has a selective tumorigenic effect on LM, inducing deregulation of “oncomirs” and tumor suppressor miRNAs, produced by NF-κB activation with molecular and early histopathological alterations linked to neoplastic transformation. Systematic acid suppression may in part convey a protective role.

show abstract

Section: Discussionmentioning

confidence: 93%

miR-21, miR-155, miR-192, and miR-375 Deregulations Related to NF-kappaB Activation in Gastroduodenal Fluid–Induced Early Preneoplastic Lesions of Laryngeal Mucosa In Vivo

Sasaki¹,

Vageli²

2016

Neoplasia

View full text Add to dashboard Cite

show abstract

“…As evidence, the role of combined pepsin and acid as a risk factor of neoplasia is supported by Adams and Heintz [41]. On the other hand, Del Negro concludes that pepsin is not carcinogenic in rats [42], while other investigators suggest that non-acidic-pepsin can induce cell proliferation in laryngopharyngeal squamous carcinoma FaDu cell lines [35,38]. Whereas previous in vitro studies described the results of a single exposure of hypopharyngeal cancer cell lines to pepsin fluid, for a prolonged period of time (1 to 24 hours) at pH higher than or equal to 7.0, we selected normal human hypopharyngeal primary cells and normal hypopharyngeal keratinocytes treated by test fluids for shorter duration (10–15 min) and over repeated exposures to mimic more closely clinical LPR events.…”

Section: Discussionmentioning

confidence: 99%

The In Vitro Effect of Acidic-Pepsin on Nuclear Factor KappaB Activation and Its Related Oncogenic Effect on Normal Human Hypopharyngeal Cells

Sasaki¹,

Toman²,

Vageli³

2016

PLoS ONE

View full text Add to dashboard Cite

BackgroundExtra-esophageal carcinogenesis has been widely discussed in relation to the chronic effects of laryngopharyngeal reflux and most prominently with pepsin historically central to this discussion. With refluxate known to include gastric (pepsin) and duodenal (bile) fluids, we recently demonstrated the mechanistic role of NF-κB in mediating the preneoplastic effects of acidic-bile. However, the role of pepsin in promoting hypopharyngeal premalignant events remains historically unclear. Here, we investigate the in vitro effect of acidic-pepsin on the NF-κB oncogenic pathway to better define its potential role in hypopharyngeal neoplasia.MethodsHuman hypopharyngeal primary cells (HHPC) and keratinocytes (HHK) were repetitively exposed to physiologic pepsin concentrations (0.1 mg/ml) at pH 4.0, 5.0 and 7.0. Cellular localization of phospho-NF-κB and bcl-2 was determined using immunofluorescence and western blotting. NF-κB transcriptional activity was tested by luc reporter and qPCR. Analysis of DNA content of pepsin treated HHK and HHPC was performed using Fluorescence-activated-cell sorting assay. To explore a possible dose related effect, pepsin concentration was reduced from 0.1 to 0.05 and 0.01 mg/ml.ResultsAt physiologic concentration, acidic-pepsin (0.1 mg/ml at pH 4.0) is lethal to most normal hypopharyngeal cells. However, in surviving cells, no NF-κB transcriptional activity is noted. Acidic-pepsin fails to activate the NF-κB or bcl-2, TNF-α, EGFR, STAT3, and wnt5α but increases the Tp53 mRNAs, in both HHPC and HHK. Weakly acidic-pepsin (pH 5.0) and neutral-pepsin (pH 7.0) induce mild activation of NF-κB with increase in TNF-α mRNAs, without oncogenic transcriptional activity. Lower concentrations of pepsin at varying pH do not produce NF-κB activity or transcriptional activation of the analyzed genes.ConclusionOur findings in vitro do not support the role of acidic-pepsin in NF-κB related hypopharyngeal carcinogenesis.

show abstract

“…Since the effects of proteolytic enzymes, which are components of gastric juice, can persist for several hours, autolysis or delayed wound healing can occur, especially in the presence of local injuries that can be caused when a prosthesis is cleaned or replaced. In these cases, damage can be a direct result of acid-induced or pepsin-induced autolysis of the mucosa or can occur as a result of alterations in intercellular junctional complexes at the molecular level or as a result of abnormalities and abacterial inflammatory reactions at the immunological level [3,9,12,17,32,34,39,43,45].…”

Section: Discussionmentioning

confidence: 99%

The management of periprosthetic leakage in the presence of supra-oesophageal reflux after prosthetic voice rehabilitation

Lorenz¹,

Grieser²,

Ehrhart³

et al. 2010

Eur Arch Otorhinolaryngol

View full text Add to dashboard Cite

The objective of the study was to investigate the influence of anti-reflux medications on the management of periprosthetic leakage in laryngectomised patients with prosthetic voice rehabilitation. Sixty patients underwent laryngectomy and prosthetic voice rehabilitation. In a prospective non-randomised study, we examined the patients clinically and assessed the presence of reflux disease using 24-h dual-probe pH monitoring before and 6 months after oral anti-reflux treatment with proton pump inhibitors (PPIs). The severity of reflux, the effectiveness of anti-reflux therapy, and the clinical success of treatment were evaluated. Reflux parameters before and after anti-reflux therapy as well as the severity and incidence of periprosthetic leakage before and after PPI therapy were the main outcome measures. The absolute number of reflux events was 162.2 (±144.3) before treatment and 63.1 (±87.9) after treatment with PPIs (p = 0.031). The reflex area index score decreased from 327.1 (±419.3) without PPIs to 123.8 (±249.7) with PPIs (p = 0.0228). The mean DeMeester score was 108.3 (±85.4) before treatment and 47.4 (±61.7) after 6 months of treatment (p = 0.0557). The relative risk of periprosthetic leakage decreased to 0.5 after anti-reflux treatment. In 19 patients, leakage problems were successfully managed by rigorous treatment with PPIs. No further surgical procedures were required in these cases. Rigorous anti-reflux treatment leads to an improvement in parameters that can be assessed objectively by 24-h dual-probe pH monitoring. In the majority of patients, the symptoms associated with periprosthetic leakage can be improved or cured.

show abstract

Experimental carcinogenesis on the oropharyngeal mucosa of rats with hydrochloric acid, sodium nitrate and pepsin

Abstract: The data of the current study could not corroborate the hypothesis that gastroesophageal and pharyngolaryngeal refluxes are carcinogenic factors to the laryngopharyngeal mucosa, and more studies are necessary in the future.

Cited by 10 publications

References 32 publications

miR-21, miR-155, miR-192, and miR-375 Deregulations Related to NF-kappaB Activation in Gastroduodenal Fluid–Induced Early Preneoplastic Lesions of Laryngeal Mucosa In Vivo

miR-21, miR-155, miR-192, and miR-375 Deregulations Related to NF-kappaB Activation in Gastroduodenal Fluid–Induced Early Preneoplastic Lesions of Laryngeal Mucosa In Vivo

The In Vitro Effect of Acidic-Pepsin on Nuclear Factor KappaB Activation and Its Related Oncogenic Effect on Normal Human Hypopharyngeal Cells

The management of periprosthetic leakage in the presence of supra-oesophageal reflux after prosthetic voice rehabilitation

Contact Info

Product

Resources

About