TFF1 Expression Research Articles

Changes in expression of oestrogen regulated and proliferation genes with neoadjuvant treatment highlight heterogeneity of clinical resistance to the aromatase inhibitor, letrozole

IntroductionClinical resistance is a major factor limiting benefits to endocrine therapy. Causes of resistance may be diverse and the mechanism of resistance in individual breast cancers is usually unknown. The present study illustrates how changes in the expression of proliferation and oestrogen-regulated genes occurring during neoadjuvant treatment with the aromatase inhibitor, letrozole, may define distinctive tumour subgroups and suggest different mechanisms of resistance in clinically endocrine resistant breast cancers.MethodsPostmenopausal women with large primary oestrogen-receptor (ER)-rich breast cancers were treated neoadjuvantly with letrozole (2.5 mg daily) for three months. Clinical response was determined by ultrasound changes in tumour volume. Tumour ribonucleic acid (RNA) from biopsies taken before, after 14 days and after three months of treatment was hybridized on Affymetrix U133A chips. Changes in expression of KIAA0101, TFF3, SERPINA3, IRS-1 and TFF1 were taken as markers of oestrogen regulation and those in CDC2, CKS-2, Cyclin B1, Thymidine Synthetase and PCNA as markers of proliferation.ResultsFifteen tumours with < 50% volume reduction over three months of treatment were classified as being clinically non-responsive. Gene expression changes after 14 days of treatment with letrozole revealed different patterns of change in oestrogen regulated and proliferation genes in individual resistant tumours. Tumours could be separated into three different subgroups as follows: i) nine cases in which both proliferation and oestrogen signalling signatures were generally reduced on treatment (ii) four cases in which both signatures were generally unaffected or increased with treatment and (iii) two cases in which expression of the majority of oestrogen-regulated genes decreased whereas proliferation genes remained unchanged or increased. In 14 out of 15 tumours, RNA profiles were also available after three months of treatment. Patterns of change observed after 14 days were maintained or accentuated at three months in nine tumours but changes in patterns were apparent in the remaining five cancers.ConclusionsDifferent dynamic patterns of expression of oestrogen-regulated and proliferation genes were observed in tumours clinically resistant to neoadjuvant letrozole, thus illustrating heterogeneity of resistance and discriminating molecular sub-classes of resistant tumours. Molecular phenotyping might help to direct circumventing therapy suggesting the targeting of specific pathways in different tumour subtypes.

Prognostic and predictive value of TFF1 for adjuvant endocrine therapy in Chinese women with early ER positive breast cancer: Comparing aromatase inhibitors with tamoxifen

Factors that predict in favor of an aromatase inhibitors (AIs) over tamoxifen (TAM) in estrogen receptor (ER) breast cancer remains to be identified. We compared progesterone receptor (PR) and trefoil factor 1 (TTF1) status (+ve versus −ve) as predictive of superior effect of AI’s over tamoxifen among a total of 1973 Chinese women with early ER+ breast cancer. The expression of TFF1 was independently associated with ER and PR. However, there was no correlation with TFF1 and HER-2 expression. Treatment effect was more pronounced in the ER+/TFF1+ postmenopausal patients with a hazard ratio favoring AIs (HR = 0.397, 95%CI 0.183–0.860), but not in the PR positive cohorts (HR = 0.466, 95%CI 0.186–1.164). We suggested that AIs was better than TAM especially in the postmenopausal patients with ER+/TFF1+ breast cancer; however the clinical application of this observation still requires further prospective studies.

674 Concomitant Knockout of M1 Muscarinic Receptors (CHRM1) in Azoxymethane-Treated Chrm3 −/− Mice Nullifies Reductions in Colon Tumor Number and Size

G A A b st ra ct s (HP). Animals were analyzed 3 and 12 months after inoculation. Morphology of the fundus was examined in sections stained with H&E. Expression of MIP-2, TNF-alfa, INF-gamma, DCAMKL1, villin, activation-induced cytidine deaminase (AID), TFF2 and COX-2 genes were measured by QRT-PCR. Distribution of parietaland DCAMKL1-positive cells was analyzed immunohistochemistry. Western blots were used to assess the phosphorylation of Smad1-5-8 and STAT3. RESULTS: The mucosa of the TG-mice exhibited decreased phosphorylation of Smad1-5-8, confirming inhibition of BMP signaling. Histological analysis showed increased height, dilated glands, decreased parietal cell number and amodest increase in inflammatory cells. The TG-mice also exhibited a 2to 3-fold increase in the expression of the inflammatory genes MIP-2, INF-gamma TNF-alfa, and COX-2. Inhibition of BMP signaling led to increased expression of the gastric progenitor cell (GPC) markers DCAMKL1 and villin, and of AID and TFF2, molecules associated with gastric carcinogenesis. Infection of the TG-mice with HP, led to a dramatic increase in inflammation, characterized by enhanced cytokine gene expression and by a robust influx of inflammatory cells. HP-infected non-TG mice exhibited inflammatory changes that were less severe then those seen in the infected TG-animals. Intraepithelial neoplasia was seen in TG-mice infected for 12 months but not in age matched, non-infected TG-mice, or HP-infected non-TG-mice. HP infection also led to enhanced expression of AID, TFF2, villin and DCAMKL1 and to the phosphorylation and activation of the oncogenic molecule STAT3. CONCLUSIONS: Loss of gastric BMP signaling leads to a pro-inflammatory state, resulting in extreme responses and accelerated development of intraepithelial neoplasia with HP infection, suggesting that BMPs normally temper gastric inflammatory responses. Low BMP signaling and heightened inflammation stimulate GPCs, including increased marker expression and cell number. Thus BMPs play an important role in modulating inflammatory responses in the development of gastric cancer.

Abstract 2237: Expression of the trefoil protein TFF3 in human breast cancers

Abstract Expression of the trefoil protein TFF3 in human breast cancers The purpose of this study is to investigate expression of the trefoil factor family protein, TFF3 in a large series of human breast cancers. Trefoil factors are small secreted proteins that are expressed in mucus secreting epithelia. They are involved in mucosal protection in part by stimulation of cell motility. It has been proposed that the motogenic actions of trefoil proteins may facilitate tumor cell invasion and metastasis. Expression of TFF3 mRNA is regulated by estrogen in human breast cancer cell lines and strong association between estrogen receptor and TFF3 mRNA expression has been demonstrated in breast tumors by expression microarray analyses. A tissue microarray was constructed from 300 primary breast tumors and 76 metastatic deposits of breast tumors from patients who presented in the northern region of England between 2002 and 2003. TFF3 expression was evaluated by immunohistochemistry with an antibody raised against correctly-folded human TFF3 protein. Each sample was analyzed in duplicate. The intensity of the immunoreactions in 1,000 tumor cells was evaluated as: absent, weak, moderate or strong. The association of TFF3 expression with various clinico-pathological features and with estrogen receptor protein expression was tested statistically with SPSS software (p&amp;lt;0.01). Clear TFF3 expression is demonstrated in normal and malignant breast epithelial cells. There is no expression in stromal, endothelial or immune cells. Expression of TFF3 varied enormously between tumor samples. TFF3 expression is highest in mucinous and tubular breast adenocarcinomas. TFF3 is expressed at higher levels in lobular than in ductal breast cancers. TFF3 expression is associated strongly with estrogen receptor expression. There is a negative association between TFF3 expression and tumor grade. There is however a positive association between TFF3 expression and presence of vascular invasion and TFF3 expression is higher in metastatic breast tumor cells than in primary breast tumor cells. This is the first report of TFF3 protein expression in a large series of human breast tumors. The strong association between TFF3 and estrogen receptor suggests that TFF3 expression is dependent on estrogen in a significant proportion of breast tumors. The high expression of TFF3 in breast tumors with vascular invasion and in metastatic breast tumor cells supports the hypothesis that TFF3 predisposes towards breast cancer cell invasion. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2237.

EGFR/HER2 inhibitor AEE788 increases ER-mediated transcription in HER2/ER-positive breast cancer cells but functions synergistically with endocrine therapy

Background:Cross-talk between receptor tyrosine kinases and the oestrogen receptor (ER) is implicated in resistance to endocrine therapy. We investigated whether AEE788 (a combined inhibitor of EGFR, HER2 and VEGFR) plus tamoxifen or letrozole enhanced the individual anti-tumour effects of these agents.Methods:Breast cancer cell lines modelling endocrine-resistant and -sensitive disease were engineered to express aromatase (A) and examined using proliferation, western blotting and ER-α transcription assays.Results:AEE788 enhanced the anti-proliferative effect of tamoxifen and letrozole in ER+ cell lines (MCF-7 2A, ZR75.1 A3 and BT474 A3). This associated with an elevated G1 arrest and nuclear accumulation of p27. It is noteworthy that AEE788 alone or in combination with endocrine therapy increased the expression of progesterone receptor (PGR) and TFF1 in BT474 A3 cells. This may indicate a mechanism of resistance to AEE788 in ER+/HER2+ breast cancers. In a ZR75.1 A3 xenograft, AEE788 alone or in combination with tamoxifen provided no further benefit compared with letrozole. However, letrozole plus AEE788 produced a significantly greater inhibition of tumour growth compared with letrozole alone.Conclusion:These data suggest that AEE788 plus letrozole in breast cancer overexpressing HER2 may provide superior anti-tumour activity, compared with single agents.

Trefoil factor 3 is induced during degenerative and inflammatory joint disease, activates matrix metalloproteinases, and enhances apoptosis of articular cartilage chondrocytes

Trefoil factor 3 (TFF3, also known as intestinal trefoil factor) is a member of a family of protease-resistant peptides containing a highly conserved motif with 6 cysteine residues. Recent studies have shown that TFF3 is expressed in injured cornea, where it plays a role in corneal wound healing, but not in healthy cornea. Since cartilage and cornea have similar matrix properties, we undertook the present study to investigate whether TFF3 could induce anabolic functions in diseased articular cartilage. We used reverse transcriptase-polymerase chain reaction, Western blot analysis, and immunohistochemistry to measure the expression of TFF3 in healthy articular cartilage, osteoarthritis (OA)-affected articular cartilage, and septic arthritis-affected articular cartilage and to assess the effects of cytokines, bacterial products, and bacterial supernatants on TFF3 production. The effects of TFF3 on matrix metalloproteinase (MMP) production were measured by enzyme-linked immunosorbent assay, and effects on chondrocyte apoptosis were studied by caspase assay and annexin V assay. Trefoil factors were not expressed in healthy human articular cartilage, but expression of TFF3 was highly up-regulated in the cartilage of patients with OA. These findings were confirmed in animal models of OA and septic arthritis, as well as in tumor necrosis factor alpha- and interleukin-1beta-treated primary human articular chondrocytes, revealing induction of Tff3/TFF3 under inflammatory conditions. Application of the recombinant TFF3 protein to cultured chondrocytes resulted in increased production of cartilage-degrading MMPs and increased chondrocyte apoptosis. In this study using articular cartilage as a model, we demonstrated that TFF3 supports catabolic functions in diseased articular cartilage. These findings widen our knowledge of the functional spectrum of TFF peptides and demonstrate that TFF3 is a multifunctional trefoil factor with the ability to link inflammation with tissue remodeling processes in articular cartilage. Moreover, our data suggest that TFF3 is a factor in the pathogenesis of OA and septic arthritis.

Synthesis and localization of trefoil factor family (TFF) peptides in the human urinary tract and TFF2 excretion into the urine

Trefoil factor family (TFF) peptides promote regeneration and repair processes of mucous epithelia. They also probably play a key role in the remarkable regenerative capacity of the urinary tract epithelia. We have localized TFF1, TFF2, and TFF3 expression systematically in surgical specimens from the urinary tract by reverse transcription with the polymerase chain reaction, Western blot analysis, and immunohistochemistry. Urine samples from patients suffering from nephrolithiasis have been investigated and compared with those of healthy controls. TFF synthesis is detectable along the entire urinary tract epithelia. TFF3 synthesis is the most pronounced followed by TFF1, whereas TFF2 synthesis is occasionally detectable but only in trace amounts. In contrast, TFF2 is the predominant TFF peptide excreted into the urine, and significantly increased urinary TFF2 levels (together with occasionally raised TFF3 levels) have been observed in patients suffering from nephrolithiasis. Thus, we consider that TFF3 plays a major part in regeneration and restitution processes in urinary tract epithelia. TFF2 and probably also TFF3 are candidate biomarkers for nephrolithiasis and possibly other inflammatory conditions of the urinary tract.

Downregulation of trefoil factor 3 gene expression in the colon of the senescence-accelerated mouse (SAM)-P6 revealed by oligonucleotide microarray analysis

Global comparison of the colonic gene expression profiles between 14-month-old senescenceaccelerated mouse (SAM)-P6 mice and SAM-R1 mice, a wild-type control, was conducted with an oligonucleotide microarray containing more than 5,000 mouse genes. Eight genes were upregulated more than two-fold and 94 genes were downregulated more than two-fold in SAM-P6 mice. The three cell defense genes intelectin1 (Itln1), trefoil factor 3 (intestinal) (Tff3) and "deleted in malignant brain tumors 1" (Dmbt1) were among those extensively downregulated. Quantitative RT-PCR analysis confirmed that Itln1 mRNA was almost undetectable in SAM-P6 colon, whereas it was readily detected in SAM-R1 colon. Colonic expression of both Tff3 and Dmbt1 mRNA was also substantially decreased, to one third and two thirds of the levels in SAM-R1 mice, respectively. A 14 kDa Tff3 dimer was detected by Western blotting in the colon of all three SAM-R1 mice, but was not present in three SAM-P6 mice. No upregulation of 3 cell defense genes was detected in 3-month-old SAM-R1 as well as SAM-P6 mice. These results suggest that a diminution of the intestinal trefoil factor system may be involved in the acceleration of aging in SAM-P6 mice.

Biological Evidence for Dual Antiangiogenic – Antiaromatase Activity of the VEGF-R Inhibitor PTK787/ZK222584.

Abstract Background: Targeting the Vascular Endothelial Growth Factor (VEGF) and Estrogen Receptor (ER) signaling pathways concomitantly may provide enhanced therapeutic benefit in ER positive breast cancer. We have previously shown the VEGFR tyrosine kinase inhibitor, PTK787/ZK222584 (PTK/ZK) is a competitive aromatase inhibitor in vitro (Banerjee et al Cancer Res 2009 69: 4716-4723). Here we investigated whether (1) PTK/ZK shows both anti-angiogenic and aromatase inhibitory properties in in vivo models of ER positive breast cancer and (2) the combination of PTK/ZK and letrozole (LET) is superior to letrozole alone.Methods: Human breast cancer cell lines were engineered to express aromatase. Both MCF7 AROM1 (ER+HER2-) and BT474 AROM3 (ER+, HER2+) cells were grown as subcutaneous xenografts with androstenedione support in ovariectomized, nude mice. Mice bearing MCF7 AROM1 or BT474 AROM3 tumors were randomised to receive daily, by oral gavage, concentrations of vehicle, PTK/ZK (25, 50, or 100mg/kg), letrozole (1 mg/kg) or PTK/ZK (50mg/kg) in combination with letrozole (1 mg/kg) for 28 days. Mice were injected with FITC-lectin prior to sacrifice at weekly intervals. Tumor sections were stained for Ki67, apoptosis (TUNEL), CD31 and PgR. Mice bearing BT474 AROM3 xenografts were also treated twice daily with either vehicle or 50 mg/kg PTK/ZK for 3 days and uteri were then weighed and TFF1 and mRNA expression determined.Results: In MCF7 AROM1 xenografts, the daily growth rate was significantly reduced in all treatment groups compared to vehicle (p&amp;lt;0.05). Whilst letrozole caused tumor regression and PTK/ZK at 50 mg/kg and 100 mg/kg stabilized tumor volumes, there was no significant differences between the tumor volumes observed in the 50 mg/kg PTK/ZK and letrozole groups (p=0.14) or letrozole and combination group (p=0.14). The cell turnover index (CTI), a composite measurement of both proliferation and apoptosis significantly decreased (&amp;gt;50%) after 1 week with all treatments (p&amp;lt;0.05) except for 25 mg/kg PTK/ZK compared to the vehicle group The addition of PTK/ZK to letrozole did not lower the CTI further compared to letrozole alone. The vessel density was significantly reduced in the PTK/ZK (50 mg/kg and 100mg/kg) and combination groups compared to the vehicle group from as early as day 7 (p&amp;lt;0.05). However, in the letrozole group, there was no significant reduction in the vessel number until day 28 (p&amp;lt;0.05). PTK/ZK also significantly decreased the growth rate and vessel number compared to the vehicle (p&amp;lt;0.05) in BT474 AROM3 xenografts. PTK/ZK decreased PgR protein expression (p&amp;lt;0.01), TFF1 expression (p=0.002) and uterine weight (p=0.02) suggesting that PTK/ZK decreases E2 signaling in vivo.Conclusions: The VEGF-R inhibitor PTK/ZK showed effects on E2-dependent gene expression consistent with aromatase inhibition as well as anti-angiogenesis in xenograft models of breast cancer. Letrozole alone showed highly effective antitumor activity and the combination of PTK/ZK with letrozole was not superior. Nonetheless these results provide further support for a potential therapeutic approach of dual inhibition of VEGF and E2 signaling using a single agent. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5139.

17: Epithelial protection and repair: the role of trefoil factor 1 in the mouse cervix through pregnancy

The cervix remodels throughout pregnancy in preparation for parturition and includes stages of softening, ripening and dilation. Understanding the molecular processes that regulate each phase is critical for identification of preterm birth risk factors due to cervical malfunction. Trefoil factor 1 (Tff1) is a member of a family of secreted proteins that play an important role in the protection and restitution of the gastrointestinal mucosal epithelium. We previously determined a temporal and robust mRNA expression of Tff1 in the pregnant cervix with maximal expression prior to birth and rapid decline postpartum.

Airway Trefoil Factor Expression during Naphthalene Injury and Repair

While the role of trefoil factors (TFF) in the maintenance of epithelial integrity in the gastrointestinal tract is well known, their involvement in wound healing in the conducting airway is less well understood. We defined the pattern of expression of TFF1, TFF2, and TFF3 in the airways of mice during repair of both severe (300 mg/kg) and moderate (200 mg/kg) naphthalene-induced Clara cell injury. Quantitative real-time PCR for tff messenger RNA expression and immunohistochemistry for protein expression were applied to airway samples obtained by microdissection of airway trees or to fixed lung tissue from mice at 6 and 24 h and 4 and 7 days after exposure to either naphthalene or an oil (vehicle) control. All three TFF were expressed in normal whole lung and airways. TFF2 was the most abundant and was enriched in airways. Injury of the airway epithelium by 300 mg/kg naphthalene caused a significant induction of tff1 gene expression at 24 h, 4 days, and 7 days. In contrast, tff2 was decreased in the high-dose group at 24 h and 4 days but returned to baseline levels by 7 days. tff3 gene expression was not significantly changed at any time point. Protein localization via immunohistochemistry did not directly correlate with the gene expression measurements. TFF1 and TFF2 expression was most intense in the degenerating Clara cells in the injury target zone at 6 and 24 h. Following the acute injury phase, TFF1 and TFF2 were localized to the luminal apices of repairing epithelial cells and to the adjacent mesenchyme in focal regions that correlated with bifurcations and the bronchoalveolar duct junction. The temporal pattern of increases in TFF1, TFF2, and TFF3 indicate a role in cell death as well as proliferation, migration, and differentiation phases of airway epithelial repair.

INOS‐derived nitric oxide mediates the increase in TFF2 expression associated with gastric damage: role of HIF‐1

Trefoil (TFF) peptides are involved in gastrointestinal mucosal restitution. An hypoxia inducible factor 1 (HIF-1)-dependent induction of TFF genes has been reported in gastric epithelial cells. Nitric oxide (NO) is associated with mucosal damage and modulates HIF-1 activity. The aim of the present study was to analyze the role of iNOS-derived NO in HIF-1alpha stabilization and TFF gene expression in damaged gastric mucosa. Aspirin caused gastric injury that peaked 6 h after dosing and returned to normality at 24 h. iNOS mRNA expression occurs in the corpus in parallel with damage. Blockade of iNOS activity did not modify gastric lesions induced by aspirin but delayed mucosal healing. Aspirin induced HIF-1alpha stabilization and TFF2 mRNA up-regulation in the mucosa, but these effects were diminished when iNOS activity was inhibited. Results obtained using a coculture setup showed that iNOS-derived NO from activated macrophages induced HIF-1alpha stabilization, TFF gene expression, and accelerated wound healing in cultured epithelial cells. Finally, transient silencing of endogenous HIF-1alpha in epithelial cells significantly undermined activated macrophage-induced TFF gene expression. Evidence suggests that the iNOS-derived NO associated with NSAID-induced gastric injury is implicated in mucosal restitution via the HIF-1-mediated induction of TFF genes.

VP-128, a novel oestradiol-platinum(II) hybrid with selective anti-tumour activity towards hormone-dependent breast cancer cells in vivo

We have previously reported the synthesis of VP-128, a new 17beta-oestradiol (E(2))-linked platinum(II) hybrid with high affinity for oestrogen receptor alpha (ERalpha). In the present study, we have investigated the anti-tumour activity of VP-128 towards breast cancer cells in vitro and in vivo. We used human ERalpha-positive (MCF-7) and -negative (MDA-MB-468) cells as a model for treatment with increasing doses of VP-128, cisplatin or E(2) in vitro and for xenograft experiments in nude mice in vivo. Compared with cisplatin, VP-128 showed markedly improved in vitro and in vivo anti-tumour activity towards ERalpha-positive MCF-7 breast cancer cells, without increased systemic toxicity. In these caspase-3-deficient cells, treatment with VP-128 overcame weak cellular sensitivity to cisplatin in vitro and in vivo. In these cells, only the hybrid induced apoptosis in an ERalpha-dependent manner, inactivated both X-linked inhibitor of apoptosis protein and Akt, and induced selective nuclear accumulation of ERalpha and the expression of ER-regulated genes c-myc and tff1, which was blocked by ERalpha-specific antagonist ICI 282 780. In the case of ERalpha-negative MDA-MB-468 cells, VP-128, but not cisplatin, induced nuclear accumulation of apoptosis-inducing factor and inhibited c-myc expression. However, VP-128 did not show enhanced in vivo anti-tumour activity compared with cisplatin. These results reveal two different modes of action for VP-128 in ERalpha-positive and -negative breast cancer cells, and highlight the promising therapeutic value of this unique E(2)-platinum hybrid for selective targeting of hormone-dependent cancers.

Intrauterine Growth Restriction Alters Postnatal Colonic Barrier Maturation in Rats

Intrauterine growth restriction (IUGR) is a leading cause of perinatal mortality and morbidity and increases the risk for necrotizing enterocolitis. We hypothesized that colonic barrier disruption could be responsible for intestinal frailty in infants and adults born with IUGR. Mucins and trefoil factor family 3 (TFF3) actively contribute to epithelium protection and healing. Our aim was to determine whether IUGR affects colonic mucosa maturation. IUGR was induced by dietary protein restriction in pregnant dams. Mucins and Tff3 expression and morphologic maturation of the colonic mucosa were followed during postnatal development of the offspring. Before weaning, mucin 2 and Tff3 protein levels were reduced in colonic mucosa of rats with IUGR compared with controls. After weaning, expression of mucin 2 (mRNA and protein) and mucin 4 (mRNA) were lower in colonic mucosa of rats with IUGR. At the same time, IUGR was associated with a reduction of crypt depth and a higher percentage of crypts in fission. We conclude that IUGR impairs mucus barrier development and is associated with long-term alterations of mucin expression. The lack of an efficient colonic barrier induced by IUGR may predispose to colonic injury not only in neonatal life but also in later life.

T1967 iNOs-Derived No Mediates TFF2 Expression in the Damaged Mucosa of Aspirin-Treated Rats

Trefoil peptides are involved in mechanisms of defense and repair in the gastrointestinal tract. We have recently demonstrated a HIF-1 dependent induction of TFF genes by hypoxia in gastric epithelial cells but less is known about TFF genes regulation In Vivo. Nitric oxide has been shown to modulate HIF-1 activity in cellular models. AIM: To analyze the role of iNOS-derived NO on HIF-1α stabilization and the TFF2 mRNA expression in the damaged mucosa of aspirin-treated rats. MATERIALM b) significantly reduced the increase in TFF2 mRNA expression observed 3 and 6 h after aspirin administration, and delayed the fall of TFF2 mRNA expression at 24h; c) diminished HIF-1α immunostaining. CONCLUSIONS: iNOS-derived NO induces HIF-1α stabilization and TFF2 mRNA expression in the damaged mucosa of aspirin-treated rats. These results suggest that iNOS-derived NO may contribute to the recovery of the gastric mucosa by modulating the epithelial gene expression.

Relationship between TFF3 expression and gastric cardiac carcinogenesis

Relationship between TFF3 expression and gastric cardiac carcinogenesis

Intestinal Expression of TFF and Related Genes During Postnatal Development in a Piglet Probiotic Trial

Trefoil factor family (TFF) peptides provide protective and reparative effects by enhancing epithelial integrity and promoting mucosal restitution. TFF peptide expression is induced after mucosal damage. These processes are of central physiological relevance during the postnatal intestinal development and are strongly influenced during the weaning period. In piglets, weaning at early maturation stages frequently causes mucosal inflammation. The aim of this study was to evaluate postnatal intestinal TFF expression in a piglet probiotic trial. Low intestinal TFF2 expression was measured at early maturation stages. Weaning, however, was associated with a distinct response of increased TFF2 expression, indicating an important role in enhancing mucosal integrity. In the distal jejunum and ileum weaning could as well be associated with increased TFF3 mRNA levels. Differential TFF1 expression was not detected. Furthermore, TFF2 localization studies in different intestinal loci were performed by means of immunohistochemistry. Expression of selected genes (TGFA, EGFR, Cox-2) known to promote TFF signaling showed differential expression pattern as well, thereby providing further functional background. Furthermore, the expression patterns of EGFR observed in this study contribute to an advanced view of previous findings of EGFR regulation mainly obtained in rodents. An upregulated EGFR expression during early postnatal development suggests a local relevance to porcine intestinal maturation. However, a feed supplementation with the probiotic strain Enterococcus faecium did not influence TFF expression.

Altered expressions of trefoil peptides genes in squamous cell carcinoma of the larynx

Aim. Trefoil peptides (TFF) are secretory products of mucin-producing cells and influence the rheological properties of mucous gels.1 They are thought to be important for the repair of mucosa damage.2 We hypothesised that TFF genes would be expressed in laryngeal mucosa and that this would be altered by smoking and neoplasia. Methods. The expressions of TFF1-3 were examined using 35S-dATP in situ hybridisation. Biopsies were obtained from the supraglottic mucosa of non-smokers (normal, n = 14) and smokers (smokers, n = 14) without laryngeal disease undergoing routine surgery and compared with tissue from archival squamous cell carcinoma (SCC: laryngectomy) specimens (n = 14). Autoradiographic intensity was scored (semi-quantitatively) by two independent observers and compared to positive and negative control tissue. Results. Scores are medians for each gene. TFF gene Normal (n = 14) Smokers (n = 14) SCC (n = 14) Expression was scored as 0–+++; 0 = absent, +++ strongly positive. TFF1 0 0 + TFF2 0/+ 0 + TFF3 ++ + ++ Discussion. Major expression of TFF3 and weak expression of TFF2 are found in normal laryngeal mucosa. TFF3 appears to be down-regulated in smokers and then up-regulated again in the cancer group (SCC). The significant findings are that TFF1 and 2 appear to be expressed or up-regulated in the cancer group. These findings suggest that laryngeal squamous cell carcinogenesis is associated with abnormal TFF1 and TFF2 genes expressions. References. 1 Kutta H., Steven P., Varoga D. et al. (2004) TFF peptides in the human false vocal folds of the larynx. Peptides. 25(5), 811–818 2 Taupin D. & Podolsky D.K. (2003) Trefoil factors: initiators of mucosal healing. Nat Rev Mol Cell Biol. 4(9), 721–732

Validation of a rodent model of Barrett’s esophagus using quantitative gene expression profiling

A rodent model of gastroduodenal-esophageal reflux can result in replacement of squamous esophageal mucosa with intestinal-type columnar mucosa and carcinoma. The validity of this model is debated, as it is unproven whether this mucosa is intestinal metaplasia due to reflux or represents migration of adjacent jejunal mucosa above the anastomosis. The aim of this study was to evaluate the esophageal intestinal-type mucosa in these animals by measuring expression of trefoil factor genes (TFF-1, -2, -3) and comparing it with adjacent jejunum in order to determine its etiology. Twenty-five rats underwent esophagojejunostomy at the ligament of Treitz to induce reflux of gastric and duodenal contents. The animals were sacrificed at 16 weeks (n = 14) and 30 weeks (n = 11). After sacrifice, the distal esophagus, jejunum, and colon were obtained. RNA was isolated, reverse transcribed, and messenger RNA (mRNA) expression of TFF-1, -2, and -3 was measured with real-time polymerase chain reaction (PCR). Linear discriminant analysis classified samples based on gene expression. Esophageal intestinal-type mucosa was present at sacrifice in 18 animals. Compared to jejunum, the expression of TFF-1 and TFF-2 mRNA in the intestinal mucosa of the distal esophagus was increased (p = 0.0007 and p < 0.0001, respectively). Expression of TFF-3 was also increased in esophageal intestinal mucosa compared with jejunum (p = 0.0002), but there was significant overlap in expression between these tissues for this gene. Linear discriminant analysis misclassified esophageal intestinal-type mucosa as jejunum in only one case. In no cases was jejunum misclassified as esophageal intestinal-type mucosa. The gene expression profile of esophageal intestinal-type mucosa following surgically induced reflux in a rodent model indicates that this represents intestinal metaplasia, not proximal migration of jejunum. This validates this model for studying the pathogenesis of Barrett's esophagus. Use of this model has potential for assessment of the impact of various therapies on the natural history of reflux disease.

Relationship between expression of CDK1,TFF2 and HPV16/18 infection in uterine cervical Carcinomas

Objective To investigate the relationship between expression of CDK1,TFF2 and HPV16/18 infection in uterine cervical carcinomas.Methods Formalin-fixed,paraffin-embedded samples including 50 cases of squamous cell carcinomas,20 cases of cervical intraepithelial neoplasia CINⅡ-Ⅲand 20 cases of CINI and 20 cases of normal cervical tissues were studied.Using Polymer DetectionSystem For Immuno-Histological Staining(PV-9000)as immunohisto-chemical detecting method.the expression of TFF2 and CDK1 in these samples were examined.The infections of HPV type 16,18 DNA were determined by PCR.Results There were significant differences for CDK1 between carcinomas,CIN and normal cervices,respectively(all P＜0.05).Expression of CDKl were correlated with tumor grades(P＜0.05).The positive expression of TFF2 in carcinomas was significantly lower than that in normal cervical tissue and CIN.There was significant differences amongthem(P＜0.05).Expression of TFF2 were correlatedwith age(P＜0.05).Thepositive rate of HPV16/18 was significant difference between cervical carcinomas,CIN and normal tissues(P＜0.05),and it was not associated with age,tumor grades,clinical stages and lymph node metastasis(all P＞0.05).The expression of CDK1 was not associated with TFF2 and HPV16/18(all P＞0.05).The expression of HPV16/18 was associated with TFF2(rs=-0.500,P＜0.05).Conclusions The results suggest that in tissues of CIN and cervical carcinomas in TFF2 and CDK1 expression changes and HPVl6/18 infection interact with each other,which can effect the progression of CIN and carcinogenesis of cervical carcinomas. Key words: Cervix neoplasms; Cyclin-dependent kinase; Peptides; Papillomavirus,human; Papillomavirus infections