Aspectos histológicos, genéticos y moleculares de las lesiones preneoplásicas de la mucosa gástrica / Histological, genetic and molecular aspects of preneoplastic lesions of the gastric mucosa

Siendo el cáncer gástrico la 3ª causa de muerte por cáncer en Chile, y existiendo estrategias de tamizaje consistentes en pesquisa de lesiones preneoplásicas de la mucosa gástrica, es relevante conocer los aspectos genéticos y moleculares que puedan ser aplicados, en la optimización de dichas estrategias a grupos de mayor riesgo. El objetivo de este manuscrito fue revisar la evidencia actual en los aspectos señalados, y de la inmunohistoquímica de 4 marcadores (p53, CDX2, MUC2 y S100A9) en la mucosa gástrica normal y en las lesiones preneoplásicas de la misma.

SUMMARY: Since gastric cancer is the 3rd leading cause of death from cancer in Chile, and there are screening strategies consisting of screening for preneoplastic lesions of the gastric mucosa, it is important to know certain genetic and molecular aspects that can be applied in optimizing these strategies for higher risk groups. The aim of this manuscript was to review the current evidence on the aforementioned aspects, and on the immunohistochemistry of 4 markers (p53, CDX2, MUC2 and S100A9) in normal gastric mucosa and in its preneoplastic lesions.

Effects of 17β-Estradiol on Colonic Permeability and Inflammation in an Azoxymethane/Dextran Sulfate Sodium-Induced Colitis Mouse Model

BACKGROUND/AIMS: Intestinal barrier dysfunction is a hallmark of inflammatory bowel diseases (IBDs) such as ulcerative colitis. This dysfunction is caused by increased permeability and the loss of tight junctions in intestinal epithelial cells. The aim of this study was to investigate whether estradiol treatment reduces colonic permeability, tight junction disruption, and inflammation in an azoxymethane (AOM)/dextran sodium sulfate (DSS) colon cancer mouse model. METHODS: The effects of 17β-estradiol (E2) were evaluated in ICR male mice 4 weeks after AOM/DSS treatment. Histological damage was scored by hematoxylin and eosin staining and the levels of the colonic mucosal cytokine myeloperoxidase (MPO) were assessed by enzyme-linked immunosorbent assay (ELISA). To evaluate the effects of E2 on intestinal permeability, tight junctions, and inflammation, we performed quantitative real-time polymerase chain reaction and Western blot analysis. Furthermore, the expression levels of mucin 2 (MUC2) and mucin 4 (MUC4) were measured as target genes for intestinal permeability, whereas zonula occludens 1 (ZO-1), occludin (OCLN), and claudin 4 (CLDN4) served as target genes for the tight junctions. RESULTS: The colitis-mediated induced damage score and MPO activity were reduced by E2 treatment (p < 0.05). In addition, the mRNA expression levels of intestinal barrier-related molecules (i.e., MUC2, ZO-1, OCLN, and CLDN4) were decreased by AOM/DSS-treatment; furthermore, this inhibition was rescued by E2 supplementation. The mRNA and protein expression of inflammation-related genes (i.e., KLF4, NF-κB, iNOS, and COX-2) was increased by AOM/DSS-treatment and ameliorated by E2. CONCLUSIONS: E2 acts through the estrogen receptor β signaling pathway to elicit anti-inflammatory effects on intestinal barrier by inducing the expression of MUC2 and tight junction molecules and inhibiting pro-inflammatory cytokines.

Effect of ceftriaxone on the intestinal epithelium and microbiota in neonatal mice / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of ceftriaxone on the intestinal epithelium and microbiota in mice in the early-life stage, as well as the recovery of the intestinal epithelium and reconstruction of intestinal microbiota in adult mice.</p><p><b>METHODS</b>A total of 36 BALB/C neonatal mice were randomly divided into control group and experimental group, with 18 mice in each group. The mice in the experimental group were given ceftriaxone 100 mg/kg every day by gavage within 21 days after birth. Those in the control group were given an equal volume of normal saline by gavage. Immunohistochemistry was used to measure the expression of Ki67, Muc2, and ZO-1 in the intestinal epithelium. qPCR and next-generation sequencing were used to analyze the overall concentration and composition of fecal bacteria.</p><p><b>RESULTS</b>After 21 days of ceftriaxone intervention, the experimental group had a significant reduction in body weight, a significant reduction in the expression of Ki67 and ZO-1 and a significant increase in the expression of Muc2 in intestinal epithelial cells, a significant reduction in the overall concentration of fecal bacteria, and a significant increase in the diversity of fecal bacteria compared with the control group (P<0.05). Firmicutes was the most common type of fecal bacteria in the experimental group, and there were large amounts of Staphylococcus and Enterococcus. The experimental group had a certain degree of recovery of the intestinal epithelium, but there were still significant differences in body weight and the structure of intestinal microbiota between the two groups at 56 days after birth (P<0.05).</p><p><b>CONCLUSIONS</b>Early ceftriaxone intervention significantly affects the development of the intestinal epithelium and the construction of intestinal microbiota in the early-life stage. The injury of the intestinal microbiota in the early-life stage may continue to the adult stage and affect growth and development and physiological metabolism.</p>

Role of using two-route ulinastatin injection to alleviate intestinal injury in septic rats / 中华创伤杂志(英文版)

PURPOSE@#Early application of protease inhibitors through the intestinal lumen could increase survival following experimental shock by blocking the pancreatic digestive enzymes. Hence, it was hypothesized that two-route injection (intraintestinal + intravenous) of ulinastatin (UTI), a broad-spectrum protease inhibitor, could better alleviate intestinal injury than single-route injection (either intravenous or intraintestinal).@*METHODS@#A sepsis model induced by lipopolysaccharide on rats was established. The rats were randomly divided into five groups: sham, sepsis, UTI intravenous injection (Uiv), UTI intraintestinal injection (Uii), and UTI intraintestinal + intravenous injection (Uii + Uiv) groups. The mucosal barrier function, enzyme-blocking effect, levels of systemic inflammatory cytokines, and 5-day survival rate were compared among groups. The small intestinal villus height (VH), crypt depth (CD), and two components of mucosal barrier (E-cadherin and mucin-2) were measured to evaluate the mucosal barrier function. The levels of trypsin and neutrophil elastase (NE) in the intestine, serum, and vital organs were measured to determine the enzyme-blocking effect.@*RESULTS@#Compared with the single-route injection group (Uiv or Uii), the two-route injection (Uii + Uiv) group displayed: (1) significantly higher levels of VH, VH/CD, E-cadherin, and mucin-2; (2) decreased trypsin and NE levels in intestine, plasma, and vital organs; (3) reduced systemic inflammatory cytokine levels; and (4) improved survival of septic rats.@*CONCLUSION@#Two-route UTI injection was superior to single-route injection in terms of alleviating intestinal injury, which might be explained by extensive blockade of proteases through different ways.

Evaluation of enemas containing sucralfate in tissue content of muc-2 protein in experimental model of diversion colitis / Avaliação de enemas com sucralfato no conteúdo tecidual da proteina muc-2 em colite de exclusão experimental

ABSTRACT Background: The effects of topical application of sucralfate (SCF) on the tissue content of MUC-2 protein have not yet been evaluated in experimental models of diversion colitis. Aim: To measure the tissue content of MUC-2 protein in the colonic mucosa diverted from fecal stream submitted to the SCF intervention. Methods: Thirty-six rats underwent derivation of intestinal transit through proximal colostomy and distal mucous fistula. The animals were divided into three groups which were submitted application of enemas with saline, SCF 1 g/kg/day and SCF 2 g/kg/day. Each group was divided into two subgroups, according to euthanasia was done after two or four weeks. The colitis diagnosis was established by histopathological study and the inflammatory intensity was evaluated by previously validated scale. The MUC-2 protein was identified by immunohistochemistry and the tissue content was measured computerized morphometry). Results: The application of enemas with SCF in the concentration of 2 g/kg/day reduced inflammatory score of the segments that were diverted from fecal stream. The content of MUC-2 in diverted colon of the animals submitted to the intervention with SCF, independently of intervention period and the used concentration, was significantly greater than animals submitted to the application of enemas containing saline (p< 0.01). The content of MUC-2 after the intervention with SCF in the concentration of 2 g/kg/day was significantly higher when compared to the animals submitted to the application containing SCF at concentration of 1.0 g/kg/day (p<0.01). The tissue content of MUC-2 reached the highest values after intervention with SCF in the concentration of 2 g/kg/day for four weeks (p<0.01). Conclusion: The preventive application of enemas containing SCF reduces the inflammatory score and avoids the reduction of tissue content of MUC-2, suggesting that the substance is a valid therapeutic strategy to preserve the mucus layer that covers the intestinal epithelium.

RESUMO Racional: Os efeitos da aplicação tópica de sucralfato (SCF) no conteúdo tecidual da proteína mucina-2 (MUC-2) ainda não foram avaliados em modelos experimentais de colite de exclusão. Objetivo: Mensurar o conteúdo tecidual da proteína MUC-2 na mucosa cólica sem trânsito intestinal submetida à intervenção com SCF. Método : Trinta e seis ratos foram submetidos à derivação intestinal por colostomia proximal terminal e fístula mucosa distal. Foram divididos em três grupos segundo recebessem clisteres contendo solução fisiológica (SF), SCF 1 g/kg/dia e SCF 2 g/kg/dia. Cada grupo foi dividido em dois subgrupos, segundo a eutanásia ser realizada após duas ou quatro semanas. O diagnóstico de colite foi estabelecido por estudo histopatológico e a intensidade inflamatória foi avaliada por escala validada. A expressão tecidual da MUC-2 foi identificada por imunoistoquímica e seu conteúdo mensurado por morfometria computadorizada. Resultados: A aplicação de clisteres com SCF na concentração de 2 g/kg/dia reduziu a intensidade inflamatória no cólon sem trânsito fecal. O conteúdo tecidual de MUC-2 no cólon sem trânsito dos animais submetidos à intervenção com SCF, independente do tempo de intervenção e da concentração utilizada, foi maior quando comparado aos animais tratados com SF (p<0,01). O conteúdo de MUC-2 após a intervenção com SCF na concentração de 2 g/kg/dia foi maior quando comparado aos animais submetidos à intervenção com concentração menor (p<0,01). O conteúdo de MUC-2 foi maior após intervenção com SCF na concentração de 2 g/kg/dia por quatro semanas (p<0,01). Conclusão: A aplicação preventiva de clisteres com SCF reduz o grau de inflamação e preserva o conteúdo tecidual de MUC-2, em segmentos desprovidos de trânsito intestinal, mostrando-se uma estratégia terapêutica válida para preservar a camada de muco que recobre o epitélio intestinal.

Effect on Muc2 gene knockdown in Ht29 cells by CRISPR/Cas9 on probiotics-mediated inhibition of E.coli K1 adhesion and invasion / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of Lactobacillus rhamnosus GG (LGG) for inhibiting E.coli K1 (E44) adhesion and invasion of an intestinal epithelial cell model with Muc2 gene knockdown established using CRISPR-Cas9 system.</p><p><b>METHODS</b>Two 20-25 bp sgRNAs targeting Muc2 were chemically synthesized to construct CRISPR expression vectors for transfection in wild-type human colonic cancer cell line Ht29. The efficiency of Muc2 knockdown was determined using Western blotting. After assessment of the viability and proliferation of the transfected cells with MTT assay, we evaluated the effects of the probiotics against E44 adhesion and invasion of the cells through a competitive exclusion assay.</p><p><b>RESULTS</b>Transfection of the cells with Lenticrisprv2 plasmid vectors resulted in a cell line with stable Muc2 knockdown by 81%. The inhibitory effects of probiotics against E44 adhesion and invasion of the transfected cells were markedly attenuated, and the relative adhesion and invasion rates of E44 were 72.23% (P<0.05) and 81.49% (P<0.05), respectively.</p><p><b>CONCLUSION</b>Muc2 knockdown causes attenuation of the inhibitory effects of probiotics against E44 adhesion and invasion of the intestinal epithelial cells, suggesting that up-regulation of Muc2 may serve as an important mechanism for the probiotics to reinforce the intestinal barrier and antagonize the pathogenic bacteria, which sheds light on a new strategy for prevention and treatment of bacterial intestinal infections.</p>

MUC1-2VNTR DNA Vaccine Induces Immune Responses in Mouse Model with Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the humoral and cellular immune responses induced by MUC1-2VNTR DNA vaccine in multiple myeloma (MM) tumor-bearing mice.</p><p><b>METHODS</b>In vitro, multiple myeloma cells were transfected by plasmid pcDNA3.1-2VNTR/myc-hisB with Lipofectamine2000. The above-mentioned mouse myeloma cells were inoculated subcutaneously into female BALB/c mice for establishing tumor-bearing animal models. These female BALB/c mice were immunized with pcDNA-2VNTR/myc-hisB or pcDNA/myc-hisB. The cytotoxic T lymphocyte (CTL) activity was detected by the LDH method and the spleen lymphocyte proliferation activity was detected by CCK-8 method.</p><p><b>RESULTS</b>After immunization of BALB/c tumor-bearing mice with recombinant plasmid for 25 days, the tumor mass (0.5605 ± 0.2065 g) was significantly lighter than that in the empty plasmid control group (1.521 ± 0.6985 g) (P < 0.01) and the control group (1.5315 ± 0.5425 g) (P < 0.01). The difference of tumor mass was not statislically significant between empty plasmid control group (1.521 ± 0.6985 g) and the control group (1.5315 ± 0.5425 g) (P > 0.05). The CTL and NK cell activity was significantly higher in the group of intramuscular injection with recombinant plasmid than that in control group. The spleen lymphocyte proliferation was statistically significantly increased after being immunized with recombinant plasmid pcDNA3.1-2VNTR/myc-hisB, compared with empty vector (P < 0.01). The results showed that MUC1-2VNTR gene immunization could induce anti-tumor effect in MM tumor-bearing mice.</p><p><b>CONCLUSION</b>MUC1-2VNTR DNA immunization can elicit both humoral and cellular tumor specific immune response to multiple myeloma in MM tumor-bearing mice. It suggested that the MUC1-2VNTR DNA vaccine may be a potential treatment measure for patients with MM.</p>

CDX-2, MUC-2 and B-catenin as intestinal markers in pure mucinous carcinoma of the breast

BACKGROUND: Pure mucinous adenocarcinoma of the breast is a rare entity characterized by the production of variable amounts of mucin comprising 1% to 6% of breast carcinomas. Some mucinous adenocarcinomas have shown expression of intestinal differentiation markers such as MUC-2. This study examines the expression of intestinal differentiation markers in this type of breast carcinoma. RESULTS: Twenty-two cases of pure mucinous adenocarcinoma of the breast were assessed. Immunochemistry was performed for beta-catenin, CDX-2 and MUC-2. All cases were positive for B-catenin. MUC-2 positivity was observed in all cases; 63. 6% were 3 plus positive. All cases were negative for CDX-2. CONCLUSIONS: These results suggest that mucinous breast carcinomas express some markers of intestinal differentiation, such as MUC-2 and beta-catenin; however, future studies with a larger series of cases and using molecular techniques that help affirm these results are needed.

A Case of Primary Retroperitoneal Mucinous Cystadenoma Arising from the Retropancreatic Area / 대한소화기학회지

Primary retroperitoneal mucinous cystadenoma is an extremely uncommon tumor, even though mucinous cystadenoma often develops in the ovary and less frequently in the pancreas. A 21-year-old female was admitted to our hospital due to severe abdominal pain. A well-demarcated, oval shaped cystic tumor at the retropancreatic area with displacement of the pancreas and surrounding major vessels was observed on CT and MRI. Exploratory laparotomy was performed, and complete excision of the entire cyst was performed without complication. The pathologic finding was consistent with primary retropancreatic mucinous cystadenoma. To the best of our knowledge, this report is the first to describe a case of retropancreatic mucinous cystadenoma arising from the retropancreatic area in Korea.

Protective effect of MUC2 siRNA against colon cancer growth through the induction of apoptosis

Mucinous colorectal cancers are highly aggressive phenotype presenting with more advanced disease and a poor prognosis. The biological mechanisms involved are unclear, but appear to be linked to mucin glycoprotein overexpression like MUC2. While the role of MUC2 in colon cancer metastasis is established, the biological events and molecular pathways modulated by MUC2 are still unknown. In this study, mucin expressing human colon cancer cells LS174T were grown 'in vitro' and MUC2 expression was inhibited by MUC2 small interfering RNA molecules [siRNA]. Cell culture and soft agar growth were measured to determine the overall effect on viability. Apoptosis was investigated by measuring protein level of polyADP-ribose polymerase [PARP], caspases-3 and -8. Finally, in vivo LS174T xenografts where grown in nude mice, different treatments included MUC siRNA, scramble siRNA or saline where administered, via tail vein injection, twice a week for two weeks. Results showed that upon treatment with MUC2-siRNA there was a 5-fold reduction cell culture growth and 9-fold reduction in soft agar growth in LS174T cells. A 3 to 5-fold increase in apoptosis was mediated by caspase-8 activation. Systemic administration of MUC2 siRNA markedly inhibited tumor growth in colon cancer xenografts grown in nude mice. Tumor growth inhibition was 59% by comparing MUC2 siRNA treatment and control siRNA treatments. There was no significance difference in tumor growth between control siRNA and normal saline treatment groups [p>0.05]. We conclude that MUC2 expression appears to protect LS174T colon cancer cells from apoptosis through extrinsic apoptosis pathway. We hypothesize that contrary to previous notions that MUC2 is a secreted glycoprotein involved in digestion and gastrointestinal tract lubrication, it appears to be involved in maintenance of LS174T cell viability. MUC2 may represent a therapeutic target in mucinous colorectal carcinomas

Role of MUC2 gene in the regulation of rat intestinal barrier function by probiotics / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate MUC2 expression in rat colons induced by probiotics and its effects on the inhibition of E.coli K1 (E44) penetration of the intestinal barrier by probiotics.</p><p><b>METHODS</b>SD rats were subjected to intragastric administration of probiotics, E44, or probiotics +E44 on a daily basis for 7 days, and MUC2 expression in the colons was determined by RT-PCR. MUC2-targeted shRNA (shRNA MUC2) and scrambled shRNA plasmids (shRNA NC) were respectively transfected into Lovo cells, and the efficiency of MUC2 knockdown was determined using qRT-PCR. Competitive exclusion assay was used to evaluate the effects of the probiotics against E44 adhesion and invasion.</p><p><b>RESULTS</b>Intestinal MUC2 mRNA expression was up-regulated in the rats after intragastric administration of probiotics, while E44 administration caused significantly lowered MUC2 expression. MUC2 expression was down-regulated (by 66.7%) by transfection with shRNA MUC2 in Lovo cells as compared with the negative control and mock control cells. The inhibition of E44 adherence and invasion by probiotics was significantly attenuated in transfected Lovo cell culture (in which the relative adhesion and invasion rates of E44 were 56.64% and 66.64%, respectively) as compared with those in the control group.</p><p><b>CONCLUSION</b>The up-regulation of MUC2 in rat colons can be one of the mechanisms of the probiotics in antagonizing the translocation of the pathogenic bacteria. Silencing MUC2 expression causes attenuated inhibitory effect of the probiotics on E. coli K1 penetration across human intestinal epithelial cells.</p>

An Inverse Relationship between the Expression of the Gastric Tumor Suppressor RUNX3 and Infection with Helicobacter pylori in Gastric Epithelial Dysplasia

BACKGROUND/AIMS: This study was performed to determine the association between RUNX3 expression and Helicobacter pylori infection in premalignant gastric lesions. METHODS: We examined 107 patients with gastric epithelial dysplasia who had undergone endoscopic mucosal resection or submucosal dissection. All tissue samples were evaluated by RUNX3 staining and subclassified by immunophenotype. H. pylori infection in dysplastic lesions and the normal surrounding tissue was examined by silver staining, and cagA status was assessed by polymerase chain reaction. RESULTS: The loss of RUNX3 expression was observed in 62 cases (57.9%), and an association with H. pylori infection was found in 54 cases (50.5%). The infection rate with the cagA-positive H. pylori strain was 63.0%. In RUNX3-negative lesions, the rate of H. pylori infection (p=0.03) and the frequency of category 4 lesions (according to the revised Vienna classification) were high (p=0.02). In addition, the gastric mucin phenotype was predominant. In RUNX3-negative category 4 lesions, the rate of cagA-positive H. pylori infection rate was high but not significantly increased (p=0.08). CONCLUSIONS: Infection with H. pylori is associated with inactivation of RUNX3 in early gastric carcinogenesis. This mechanism was prominent in gastric cancer with a gastric mucin phenotype.

Colloid carcinoma of pancreas: a clinicopathologic study of 4 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the pathologic features, diagnosis, differential diagnosis and molecular characteristics of colloid carcinoma of the pancreas.</p><p><b>METHODS</b>The clinical findings, morphologic features, immunophenotype and K-ras gene alterations were investigated in 4 cases of pancreatic colloid carcinoma.</p><p><b>RESULTS</b>In the 4 cases of colloid carcinoma of the pancreas, three tumors were located in the head of the pancreas, one was located in the body and tail. The average age was 56.5 years old. The presenting symptom was abdominal pain in 2 cases, increased level of U-GLU in 1 patient, and an accidental finding presented in 1 patient. Grossly, 3 cases were cystic and solid, with mucin in the cyst; 1 case was solid. Microscopically, the colloid carcinoma was characterized by large pools of extracellular mucin, containing neoplastic cells, which were in the pattern of cuboidal, cribriform or irregular clusters, or formed an incomplete lining separating mucin pools from the stroma. Three cases developed from pre-existing pancreatic ductal adenocarcinoma (IPMN), intestinal-type, and 1 from IPMN, pancreatobiliary-type. Immunohistochemical studies showed that MUC2 was positive in 3 cases, and MUC1 in 1 case. K-ras gene mutation was identified in 2 cases, showing a single-amino-acid substitution in codon 12, as Gly12Asp (GGT > GAT) and Gly12Arg (GGT > CGT).</p><p><b>CONCLUSIONS</b>Pancreatic colloid carcinoma is a rare variant of pancreatic ductal adenocarcinoma, which is associated with IPMN and mucinous cystic neoplasms. Positive MUC2 staining and absent MUC1 expression are commonly found, and K-ras gene mutation is occasionally identified in these tumors.</p>

Expression of mucin regulated by recombinant human interleukin-6 in HT-29 cell of colon cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study the regulation of the mucin protein in colon cancer cell line HT-29 by recombinant human interleukin-6(rIL-6) and to further elucidate the development of colon cancer.</p><p><b>METHODS</b>The HT-29 cells were treated with different concentrations of rIL-6(1, 2, 5, 10 and 20 μg/L), then flow cytometry and RT-PCR were used to detect the expression of mucin1 and mucin2. Transwell invasion assay was used to observe the effect of invasion capability of rIL-6 to HT-29 cells.</p><p><b>RESULTS</b>In colon cancer, the expression of mucin 1 could be promoted by rIL-6 with concentration above 2 μg/L, the expression rates were(12.5±1.6)%, (26.6±2.7)%, (33.9±2.8)% and (58.9±2.5)%, respectively, higher than (8.0±0.8)% in the negative controls (P<0.01), meanwhile, the expression of mucin 2 decreased by rIL-6 with concentration above 2 μg/L, the expression rates were(30.5±2.6)%, (17.0±2.7)%, (11.0±2.0)% and (5.3±1.8)%, respectively, lower than (41.6±3.6)% in negative control(P<0.01). With the increase in rIL-6 concentration, the invasion of HT-29 cells was enhanced.</p><p><b>CONCLUSIONS</b>In colon cancer, the expression of mucin1 can be promoted by rIL-6, while the expression of mucin2 can be inhibited. IL-6 is a promoting effect factor in colon cancer invasion and metastasis.</p>

Hath1 gene transfer inhibits the proliferation of colonic cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of Hath1 gene transfer on the proliferation of colonic cancer cells in vitro.</p><p><b>METHODS</b>The recombinant plasmid pcDNA3.1(+)-Hath1 was transfected into HT29 colonic cancer cells, and 3 positive cell clones were randomly selected to test the levels of Hath1 mRNA, Muc2 mRNA, Hath1, Muc2, cyclin D1 and p27 by quantitative real-time RT-PCR and Western blotting. The proliferation of the transfected HT29 cells was observed by means of colony formation assay and xenograft growth in nude mice.</p><p><b>RESULTS</b>Hath1 significantly down-regulated of cyclin D1 and up-regulate of p27 expressions and inhibited the proliferation of HT29 cells.</p><p><b>CONCLUSION</b>Hath1 gene may be an anti-oncogene in colon carcinogenesis.</p>

Interleukin-1beta induces MUC2/MUC5B gene expression in human nasal epithelial cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To demonstrate the effects of IL-1beta on MUC2/MUC5B gene expression in cultured human nasal epithelial cells.@*METHOD@#In passage-2 cultured human nasal epithelial cells, the mRNA levels of MUC2/MUC5B gene expression induced by IL-1 beta were determined by fluorescent quantitative RT-PCR.@*RESULT@#MUC2/MUC5B mRNAs were detected after 24 h of exposure to IL-1beta. MUC2 mRNA levels in IL-1 beta treatment [(39.26 +/- 6.10) x 10(4) copy/microg] were significantly higher than control [(5.70 +/- 4.16) x 10(4) copy/microg] (P < 0.01). MUC5BmRNA levels in IL-1beta treatment [(5.7 +/- 2.06) x 10(5) copy/microg] were significantly higher than control [(1.11 +/- 0.72) 10(5) copy/microg] (P < 0.05).@*CONCLUSION@#IL-1 beta increased MUC2/MUC5B mRNA levels in human nasal epithelial cells. These results suggest that IL-1beta may enhance mucin gene expression in cultured human nasal epithelial cells.

Clinicopathologic and immunohistochemical study on early gastric signet-ring cell carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the expression of gastric and intestinal phenotypic markers in gastric signet-ring cell (SRC) carcinoma and the relationship with the clinicopathologic parameters and prognosis.</p><p><b>METHODS</b>Immunohistochemical study was carried out in 91 cases of early-stage SRC carcinoma using MUC1, MUC5AC and MUC6 antibodies as the gastric phenotypic markers and MUC2 and CDX2 antibodies as the intestinal phenotypic markers. According to the expression of phenotypic markers, the tumors were classified into three different subgroups: gastric, intestinal and mixed. The findings were analyzed together with various clinical parameters and follow-up data.</p><p><b>RESULTS</b>Amongst the 91 cases studied, 53 cases (58.2%) belonged to gastric type, 22 cases (24.2%) mixed type and 16 cases (17.6%) intestinal type. The positive rates of MUC2 and CDX2 in early submucosal carcinoma were significantly higher than those in mucosal carcinoma (P < 0.01). On the other hand, the rates of MUC5AC and MUC6 expression in early submucosal carcinoma were significantly lower than those in mucosal carcinoma (P < 0.01 and P < 0.05). The rates of MUC2 and CDX2 expression in cases with lymph node metastasis and vascular invasion were significantly higher than those in cases without nodal or vascular involvement (P < 0.05). The expression of CDX2 was also significantly higher in cases with larger tumor size (P < 0.05). Cases with intestinal phenotype more likely had invasion deeper than mucosal layer and carried higher chance of lymph node metastasis (P = 0.000 and P = 0.003). Intestinal and mixed types correlated with shortened five-year survival.</p><p><b>CONCLUSIONS</b>The intestinal type of SRC carcinoma is associated with poorer biologic behavior and prognosis, as compared with that of the gastric type. Classification on the basis of immunophenotypic markers may be useful in predicting prognosis and guiding treatment for patients with gastric SRC carcinoma.</p>

The expression of mucins gene in the human nasal polyps and allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To detect the mucin gene (MUC2, MUC5AC, MUC5B, MUC18 and MUC19) expression in the nasal polyps, allergic rhinitis (AR) and the normal nasal mucosa in human. To investigate the role and clinical significance of mucin gene in the pathogenesis of nasal polyps and AR patients.@*METHOD@#We obtained samples from 35 cases of nasal polyps, 18 cases of AR inferior turbinate and 18 cases of simple nasal septum deviation inferior turbinate. Specimens were analyzed with RT-PCR and Real-time FQ-RT-PCR.@*RESULT@#The results of RT-PCR and FQ-RT-PCR showed that the expression of MUC5AC, MUC5B in nasal polyps and AR patients was significantly higher than that in normal mucosa (P0.05). The expression of MUC2, MUC18 in nasal polyps and AR was not significantly different from that in normal mucosa (P>0.05). And the results of RT-PCR for MUC19 expression in AR was higher than that in nasal polyps group and normal group (P<0.05 or P<0.01).@*CONCLUSION@#MUC5AC and MUC5B are highly expressed in epithelium of human nasal polyps and AR, and they take part in mucus over-secretion in nasal polyps and AR. The expression of MUC19 in AR was higher than that in nasal polyps group and normal group. It indicates that the secretion of MUC19 in allergic rhinitis was on high level. There was no difference of the expression of MUC2 and MUC18 in nasal polyps group, AR group and in normal group.

Effects of platelet activating factor receptor antagonist in intestinal mucin-2 during endotoxemia on young rats / 中华儿科杂志

<p><b>OBJECTIVE</b>This study aimed to investigate the roles of platelet activating factor (PAF) receptor antagonist in damage to intestinal mucin-2 (MUC2) during endotoxemia on young rats.</p><p><b>METHODS</b>Eighteen-day-old Wistar rats were randomized to be treated with lipopolysaccharide (LPS) (5 mg/kg), LPS plus PAF receptor antagonist and normal saline injection (control). PAF receptor antagonist BN52021 5 mg/kg was administered 30 minutes before or after LPS injection (pretreatment group or treatment group). The ileum specimens (n = 8) were harvested at 1.5, 3, 6, 24, 48 and 72 hours after LPS injection. Transmission electron microscopy was used for morphologic evaluation. Immunohistochemistry was used to determine MUC2 in intestinal mucosa.</p><p><b>RESULTS</b>Microvilli and tight junctions were intact in the control group. Enlargement of tight junctions were seen in the LPS group and microvilli were thin, rare or disrupted, shed. The rough endoplasmic reticulum, mitochondria, and glycogen particles were injured. The changes of the pretreatment and treatment group were slightly milder than that in the LPS group. Ileum of a control rat in which a thin layer of mucus covering the epithelial surface and mucin-containing goblet cells appeared very distended. The experiment group showed a decrease or irregularly distributed membranous mucus expression. The MUC2 content of absorbance significantly decreased in the LPS challenged group compared with that in the control group (P < 0.01), and reached a nadir at 6 hours (0.1841 +/- 0.0047) vs. the control group (0.2091 +/- 0.0060) (P < 0.01). The tendency of the level of MUC2 in the pretreatment and treatment group was the same as that of the LPS group, and the level of MUC2 in the pretreatment and treatment group was higher than that in the LPS group at each time point. ANOVA analysis showed that the inter-group and intra-group difference had statistical significance (P < 0.05).</p><p><b>CONCLUSIONS</b>PAF may play some roles in the injury of intestinal mucus barrier function during endotoxemia. Preventive and remedial use of PAF receptor antagonist BN52021 may relieve intestinal injury.</p>

Mucin expression profile in Barrett's, dysplasia, adenocarcinoma sequence in the esophagus.

BACKGROUND: The molecular events that accompany the progression to adenocarcinoma (ADC) of the esophagus are poorly understood. Aberrant mucin receptor expression can contribute to increased cell growth and metastatic ability. AIM: The aim of this study was to establish a pattern for mucin (MUC) gene expression in the esophageal mucosa under normal and pathological conditions. SETTING: University Hospital Cancer Center Laboratory. Archived tissue samples studied in a retrospective fashion. MATERIALS AND METHODS: Tissue samples were obtained from the archives of patients with histological evidence of Barrett's esophagus (BE) progressing to ADC. Immunohistochemical analysis was performed using mouse monoclonal antibodies for MUC1, MUC2, MUC5AC, MUC6. Semiquantitative scoring of histological staining was performed using a linear scoring system: 0-staining absent; 1-staining in 0-25%; 2-staining in 25-50%; and 3-staining in 50-75% of the epithelium. The Binomial test was used to explore trends and differences in frequency of mucin expression along the pathological sequence. RESULTS: Only mild superficial staining of MUC1 was seen in normal squamous epithelium. MUC1 and MUC2 were uniformly expressed in all samples (7/7) of BE and dysplasia (P=0.008). MUC1 expression was upregulated (7/7) in progression to adenocarcinoma (P=0.008). The secretory mucins, MUC5AC and MUC6 showed a decrease in expression with progression from BE to dysplasia to ADC (P< 0.05). CONCLUSIONS: Downregulation of MUC5AC and MUC6 decreases mucosal protection against gastric acid. Increasing MUC1 expression is associated with progression from dysplasia to ADC. Upregulation of MUC2 reflects intestinal metaplasia in BE.