Metachronous esophageal squamous cell cancer after gastrectomy for gastric cancer

Background

The clinical and biological characteristics of metachronous esophageal squamous cell cancer (ESCC) after gastrectomy for gastric cancer have yet to be sufficiently elucidated. The aim of the present study was to examine carcinogenesis in such patients.

Methods

Subjects comprised 11 patients with metachronous carcinoma in whom ESCC occurred after gastric cancer (metachronous ESCC), 9 patients with simultaneously occurring gastric cancer and ESCC (simultaneous ESCC) and 52 patients with ESCC alone. We investigated the clinicopathological findings and biological properties using p53, p21 and cyclin D1 expression.

Results

The positive rate for the intraepithelial spread of tumor was higher for metachronous ESCC than for simultaneous ESCC (p < 0.05). The number of dysplastic lesions in metachronous ESCC, simultaneous ESCC and ESCC alone was 56, 41 and 44, respectively. The rate of positive p53 expression in dysplasia was significantly higher for metachronous ESCC than for ESCC alone (p = 0.03).

Conclusions

Positive expression of p53 was found in not only the primary tumor, but also intraepithelial neoplasia around the tumor in metachronous ESCC. Chronic gastroesophageal reflux due to gastrectomy may be involved in the process of carcinogenesis in addition to environmental and genetic factors for metachronous ESCC. Further studies of a larger number of patients with metachronous ESCC and a history of gastrectomy are warranted.     

Aberrant p16INK4a methylation is a frequent event in colorectal cancers: prognostic value and relation to mRNA expression and immunoreactivity

Purpose  

Aberrant p16^INK4a promoter methylation is common in colorectal cancer (CRC), but its clinicopathological significance remains controversial. The present study was therefore conducted to analyze p16^INK4a methylation and its relationship to clinicopathological features, mRNA levels and immunoreactivity in a series of lesions.     

Identification of <Emphasis Type="Italic">GPX3</Emphasis> Epigenetically Silenced by CpG Methylation in Human Esophageal Squamous Cell Carcinoma

Background  

Esophageal squamous cell carcinoma (ESCC) is one of the most common causes of cancer mortality in the gastrointestinal tract. Promoter hypermethylation of tumor suppressor genes contributes to gene inactivation during development of ESCC.     

The effects of vascular endothelial growth factor C knockdown in esophageal squamous cell carcinoma

Purpose  

We investigated the role of vascular endothelial growth factor C (VEGF-C) in esophageal squamous cell carcinoma (ESCC) by knocking down VEGF-C expression in the ESCC cell line EC9706.     

Esophageal Squamous Cell Carcinoma Patients Have an Increased Risk of Coexisting Colorectal Neoplasms

Background/Aims

Esophageal squamous cell carcinoma (ESCC) and colorectal neoplasms (CRNs) share risk factors. We aimed to investigate whether the CRN risk is increased in ESCC patients.

Methods

ESCC patients who underwent a colonoscopy within 1 year of diagnosis were retrospectively analyzed. Patients were matched 1:3 by age, gender, and body mass index to asymptomatic controls. CRN was defined as the histological confirmation of adenoma or adenocarcinoma. Advanced CRN was defined as any of the following: ≥3 adenomas, high-grade dysplasia, villous features, tumor ≥1 cm, or adenocarcinoma. The risk factors for both CRN and advanced CRN were evaluated by univariate and multivariate analyses.

Results

Sixty ESCC patients were compared with 180 controls. The ESCC group had significantly higher numbers of CRNs (odds ratio [OR], 2.311; 95% confidence interval [CI], 1.265 to 4.220; p=0.006) and advanced CRNs (OR, 2.317; 95% CI, 1.185 to 4.530; p=0.013). Significant risk factors for both CRN and advanced CRN by multivariate analysis included ESCC (OR, 2.157, 95% CI, 1.106 to 4.070, p=0.024; and OR, 2.157, 95% CI, 1.045 to 4.454, p=0.038, respectively) and older age (OR, 1.068, 95% CI, 1.032 to 1.106, p<0.001; and OR, 1.065, 95% CI, 1.024 to 1.109, p=0.002, respectively).

Conclusions

The rates of CRN and advanced CRN are significantly increased in ESCC. Colonos-copy should be considered at ESCC diagnosis.     

Expression of Nucleostemin, epidermal growth factor and epidermal growth factor receptor in human esophageal squamous cell carcinoma tissues

Objective  

To determine the expression of nucleostemin (NS), epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) mRNA in human esophageal squamous cell carcinoma (ESCC) tissues and their association in a human ESCC cell line.     

Chromosomal imbalances are uncommon in chagasic megaesophagus

Background  

Chagas' disease is a human tropical parasitic illness and a subset of the chronic patients develop megaesophagus or megacolon. The esophagus dilation is known as chagasic megaesophagus (CM) and one of the severe late consequences of CM is the increased risk for esophageal carcinoma (ESCC). Based on the association between CM and ESCC, we investigated whether genes frequently showing unbalanced copy numbers in ESCC were altered in CM by fluorescence in situ (FISH) technology.     

The expression and role of Aquaporin 5 in esophageal squamous cell carcinoma

Background

Aquaporins (AQPs) are water channel proteins that facilitate transcellular water movements. Recent studies have shown that AQP5 is expressed in various cancers, and plays a role in tumor progression. However, its expression and role in esophageal squamous cell carcinoma (ESCC) have not been investigated. We examined the pathophysiologic role of AQP5 in cell proliferation and survival, and also investigated its expression and effects on the prognosis of ESCC patients.

Methods

AQP5 expression in human ESCC cell lines was analyzed by Western blot testing. Knockdown experiments with AQP5 siRNA were conducted, and the effects on cell proliferation, cell cycle progression, and cell survival were analyzed. The cells’ gene expression profiles were analyzed by microarray analysis. Immunohistochemistry of AQP5 for 68 primary tumor samples obtained from ESCC patients undergoing esophagectomy was performed.

Results

AQP5 expression was high in TE2 and TE5 cells. In these cells, the knockdown of AQP5 using siRNA inhibited cell proliferation and G₁-S phase progression, and induced apoptosis. The AQP5 siRNA transfected TE5 cells showed significant increase in p21 and decrease in CCND1 mRNA expression, respectively. The expression pattern of AQP5 and p21 protein was sharply contrasted, but AQP5 and CCND1 protein expression showed a similar pattern in ESCC tissue. These findings agree with the microarray results. Immunohistochemical staining of 68 ESCC patients showed the AQP5 expression is associated with tumor size, histological type, and tumor recurrence.

Conclusion

The AQP5 expression in ESCC cells may affect cell proliferation and survival, and impact on the prognosis of ESCC patients.     

Expression and prognostic relevance of p21WAF1 in stage III esophageal squamous cell carcinoma

The prognostic effect of p21^WAF1 expression on esophageal squamous cell carcinoma patients is controversial. Further clarifying the effect of this protein is beneficial for optimizing the patient outcomes. In the current study, we investigated the expression of p21^WAF1 protein in 189 specimens of stage III ESCC by immunohistochemistry. As shown by the Kaplan–Meier curve, the overall survival rate of the positive‐expression group was significantly higher than that of the negative‐expression group (P < 0.05). No significant correlation was observed between p21^WAF1 expression and clinicopathological parameters in terms of gender, age, tumor location, tumor grade, pathological stage, and number of regional lymph node metastases (P > 0.05). We concluded that p21^WAF1 played an intricate role in the tumorigenesis and development of ESCC. p21^WAF1 could serve as a positive prognostic predictor for stage III ESCC patients.     

Deletion of p47phox attenuates the progression of diabetic nephropathy and reduces the severity of diabetes in the Akita mouse

Aims/hypothesis  

Reactive oxygen species (ROS) contribute to diabetes-induced glomerular injury and endoplasmic reticulum (ER) stress-induced beta cell dysfunction, but the source of ROS has not been fully elucidated. Our aim was to determine whether p47 ^phox -dependent activation of NADPH oxidase is responsible for hyperglycaemia-induced glomerular injury in the Akita mouse, a model of type 1 diabetes mellitus resulting from ER stress-induced beta cell dysfunction.     

p185(BCR/ABL) has a lower sensitivity than p210(BCR/ABL) to the allosteric inhibitor GNF-2 in Philadelphia chromosome-positive acute lymphatic leukemia

Background

The t(9;22) translocation leads to the formation of the chimeric breakpoint cluster region/c-abl oncogene 1 (BCR/ABL) fusion gene on der22, the Philadelphia chromosome. The p185^BCR/ABL or the p210^BCR/ABL fusion proteins are encoded as a result of the translocation, depending on whether a “minor” or “major” breakpoint occurs, respectively. Both p185^BCR/ABL and p210^BCR/ABL exhibit constitutively activated ABL kinase activity. Through fusion to BCR the ABL kinase in p185^BCR/ABL and p210^BCR/ABL “escapes” the auto-inhibition mechanisms of c-ABL, such as allosteric inhibition. A novel class of compounds including GNF-2 restores allosteric inhibition of the kinase activity and the transformation potential of BCR/ABL. Here we investigated whether there are differences between p185^BCR/ABL and p210^BCR/ABL regarding their sensitivity towards allosteric inhibition by GNF-2 in models of Philadelphia chromosome-positive acute lymphatic leukemia.

Design and Methods

We investigated the anti-proliferative activity of GNF-2 in different Philadelphia chromosome-positive acute lymphatic leukemia models, such as cell lines, patient-derived long-term cultures and factor-dependent lymphatic Ba/F3 cells expressing either p185^BCR/ABL or p210^BCR/ABL and their resistance mutants.

Results

The inhibitory effects of GNF-2 differed constantly between p185^BCR/ABL and p210^BCR/ABL expressing cells. In all three Philadelphia chromosome-positive acute lymphatic leukemia models, p210^BCR/ABL-transformed cells were more sensitive to GNF-2 than were p185BCR/ABL-positive cells. Similar results were obtained for p185^BCR/ABL and the p210^BCR/ABL harboring resistance mutations.

Conclusions

Our data provide the first evidence of a differential response of p185^BCR/ABL- and p210^BCR/ABL- transformed cells to allosteric inhibition by GNF-2, which is of importance for the treatment of patients with Philadelphia chromosome-positive acute lymphatic leukemia.     

Gender differences in clinicopathological features and prognosis of squamous cell carcinoma of the esophagus

Background

Esophageal squamous cell carcinoma (ESCC) is more frequent in male, and female ESCC patients have better prognosis and tend to diagnose at an earlier stage than male. Regarding these female advantages, gender differences of immunological reaction and sex hormone relations were investigated previously. However, the gender differences of clinicopathological features and prognostic factors of ESCC remain well unknown.

Methods

A total of consecutive 170 Japanese patients, including 28 females with ESCC who newly diagnosed and underwent esophagectomy between January 2004 and March 2013 in our institute, were examined. Clinicopathological features and p53 expression, a potent biomarker reflecting chemoresistance and prognosis, were compared. Prognostic factors were analyzed using a multivariate analysis.

Results

The rates of current drinking, flusher, smoking habits, and Brinkman index in female were lower than those in male (p < 0.001). Tumor location, tumor differentiation, T factor, N factor, clinical stage, and contents of initial treatment had no gender differences. Especially, in the population that received neoadjuvant chemotherapy, excellent pathological effectiveness (>Grade2) was seen much more in female significantly (36.1:66.7 %, p = 0.048). Immunohistostaining revealed positive rates of p53 expression were significantly high in male (50.4:30.5 %, p = 0.007). Postoperative complication occurred more frequently in male than female (52.8:28.6 %, p = 0.024). Estimated 5-year disease-specific survivals by Kaplan–Meier method were worse in male than female at rates of 46.2 and 76.7 %, respectively (p = 0.045). Multivariate analysis by Cox’s proportional hazards model showed that female gender (HR: 0.508, p = 0.023) and tumor depth (HR: 0.572, p = 0.018) were independent prognostic factors of ESCC after resection.

Conclusions

Female ESCC showed prefer prognosis to male ESCC. Low p53 imunohistochemical expression in the female ESCC patients might be related with higher sensitivity to neoadjuvant chemotherapy.

     

Epigenetic Inactivation of the SFRP1 Gene in Esophageal Squamous Cell Carcinoma

Introduction  

The secreted frizzled-related protein 1 (SFRP1) gene, as a Wnt signaling modulator, is frequently inactivated by promoter methylation in many tumors including gastric cancer, breast cancer, oral squamous cell carcinoma, and esophageal adenocarcinoma. However, the role of SFRP1 in esophageal squamous cell carcinoma (ESCC) is not clear. In this study, we investigated the epigenetic inactivation of the SFRP1 gene in ESCC.     

p12CDK2-AP1 inhibits breast cancer cell proliferation and in vivo tumor growth

Purpose

p12^CDK2-AP1 is a growth suppressor that negatively regulates cyclin-dependent kinase 2 (CDK2) activities and shows to interfere in DNA replication. Here, we aim to elucidate the role of p12^CDK2-AP1 in breast cancer progression.

Methods

Expression of p12^CDK2-AP1 protein was examined in 60 pairs of breast cancer specimens and adjacent non-tumor tissues using immunohistochemistry assay. Loss-of-function and gain-of-function analysis was performed on MCF-7 and MDA-MB-231 breast cancer cells. Routine assays including MTT, colony formation, flow cytometry, and tumorigenesis in nude mice were performed and cell cycle regulators were analyzed.

Results

p12^CDK2-AP1 was found to be significantly downregulated in 60 breast cancer tissues compared to corresponding non-tumorous tissues. The proliferation and colony formation ability was inhibited in cells that transduced with p12^CDK2-AP1 over-expression lentivirus, but enhanced in cells that transduced with p12^CDK2-AP1 RNAi lentivirus. p12^CDK2-AP1 over-expression led to G0/G1 phase arrest in the cell cycle and caused expression changes of cell cycle-related genes (CDK2, CDK4, p16^Ink4A, p21^Cip1/Waf1). Furthermore, p12^CDK2-AP1 over-expression inhibited in vivo tumor growth in immunodeficiency mice, supporting an inhibitory role for p12^CDK2-AP1 in breast cancer development.

Conclusions

As a cell cycle regulator, p12^CDK2-AP1 is involved in the development of breast cancer and maybe a potential therapeutic candidate to suppress tumorigenicity in breast cancer.     

A pilot study of nimotuzumab combined with cisplatin and 5-FU in patients with advanced esophageal squamous cell carcinoma

Objective

To observe the short-term effect and adverse reaction of Nimotuzumab in combination with chemotherapy on advanced esophageal squamous cell carcinoma (ESCC).

Method

19 patients were treated with the following protocol: Nimotuzumab 400mg/time/week in the 1^st week, 200mg/time/week from the 2^nd to 8^th week, intravenous drip (IVD); Cisplatin 80 mg/m², IVD, 4 weeks a cycle and repeated again; 5-FU 750 mg/m², continuous 24-hours pump-in × 5 days, 4 weeks a cycle and repeated again.

Result

16 of all 19 patients can be evaluated. After treatment, RP is 42.1% (95% CI, 19.9-64.3%) and DCR is 68.4%; the main side effects include arrest of bone marrow, gastrointestinal reactions, asthenia, etc.

Conclusion

Nimotuzumab in combination with cisplatin/5-FU regimens in patients with advanced ESCC is safe and effective, which deserves a further expanded sample research.     

Retrospective evaluation of adverse events of neoadjuvant or induction chemotherapy with docetaxel, cisplatin, and 5-fluorouracil in esophageal squamous cell carcinoma

Background

Neoadjuvant or induction chemotherapy with docetaxel, cisplatin, and 5-fluorouracil (DCF) is regarded as one of the most promising regimens for locally advanced esophageal squamous cell carcinoma (ESCC), but has a high incidence of adverse events. We retrospectively evaluated adverse events in patients with ESCC who received DCF.

Methods

Between December 2009 and May 2012, 57 patients with Stage II to IV locally advanced ESCC received three cycles of DCF (docetaxel 70–75 mg/m², day 1; cisplatin 70–75 mg/m², day 1; 5-fluorouracil, 750 mg/m², days 1–5) as neoadjuvant or induction chemotherapy in a single institution. Adverse events of DCF were evaluated retrospectively.

Results

The median age of the subjects was 65 years (range 41–74). Fifty-four patients (95 %) received three cycles of DCF as scheduled. Dose modifications were made for 34 patients (60 %). There were no treatment delays or unplanned admissions. Grade 3 or 4 neutropenia developed in 53 patients (93 %), including 18 (32 %) with febrile neutropenia. Other grade 3 or 4 toxic effects were hyponatremia in 15 patients (26 %), hypokalemia in 10 (18 %), oral mucositis in 9 (16 %), diarrhea and hypocalcemia in 8 (14 %) each, fatigue in 4 (7 %), and vomiting, anorexia, hiccups, hypoalbuminemia, and elevations of alanine aminotransferase in 1 (2 %) each. No treatment-related deaths occurred within 30 days after completion of DCF.

Conclusions

For patients with locally advanced ESCC, neoadjuvant or induction chemotherapy with DCF is tolerable. DCF is a viable treatment option for fit patients with locally advanced ESCC.     

Tyr323‐dependent p38 activation is associated with rheumatoid arthritis and correlates with disease activity

Objective

The p38 MAPK is important in the pathogenic immune response in rheumatoid arthritis (RA). The p38 molecule can be activated through phosphorylation on Thr¹⁸⁰–Tyr¹⁸² by upstream MAPK kinases and via an alternative pathway through phosphorylation on Tyr³²³. We undertook this study to quantify the phosphorylation of Tyr³²³ p38 and of Thr¹⁸⁰–Tyr¹⁸² p38 on T cells from healthy controls and patients with RA or ankylosing spondylitis (AS) to identify variables associated with p38 phosphorylation and disease activity.

Methods

We measured p38 phosphorylation on Tyr³²³ and Thr¹⁸⁰–Tyr¹⁸² by flow cytometry and Western blotting on T cells from 30 control subjects, 33 AS patients, 30 patients with RA in remission, and 79 patients with active RA. We collected the clinical characteristics and analyzed correlations between clinical variables, the Disease Activity Score in 28 joints (DAS28), and p38 phosphorylation levels. Multivariate regression analysis was performed to identify variables associated with p38 phosphorylation on Tyr³²³ and Thr¹⁸⁰–Tyr¹⁸².

Results

Phosphorylation of p38 on Tyr³²³ was higher in T cells from patients with active RA (P = 0.008 versus healthy controls) than in patients with RA in remission or in patients with AS. Tyr³²³ p38 phosphorylation was associated with disease activity determined by the DAS28 (P = 0.017). Enhanced p38 phosphorylation was linked to Lck‐mediated activation of the Tyr³²³‐dependent pathway in the absence of upstream MAPKK activation.

Conclusion

Our results indicate that phosphorylation status on Tyr³²³ p38 correlates with RA disease activity and suggest that the Tyr³²³‐dependent pathway is an attractive target for down‐regulation of p38 activity in RA patients.

     

Histone deacetylase inhibitor suppression of autoantibody‐mediated arthritis in mice via regulation of p16INK4a and p21WAF1/Cip1 expression

Objective

To examine whether depsipeptide (FK228), a histone deacetylase (HDA) inhibitor, has inhibitory effects on the proliferation of synovial fibroblasts from rheumatoid arthritis (RA) patients, and to examine the effects of systemic administration of FK228 in an animal model of arthritis.

Methods

Autoantibody‐mediated arthritis (AMA) was induced in 19 male DBA/1 mice (6–7 weeks old); 10 of them were treated by intravenous administration of FK228 (2.5 mg/kg), and 9 were used as controls. The effects of FK228 were examined by radiographic, histologic, and immunohistochemical analyses and arthritis scores. RA synovial fibroblasts (RASFs) were obtained at the time of joint replacement surgery. In vitro effects of FK228 on cell proliferation were assessed by MTT assay. Cell morphology was examined by light and transmission electron microscopy. The effects on the expression of the cell cycle regulators p16^INK4a and p21^WAF1/Cip1 were examined by real‐time polymerase chain reaction and Western blot analysis. The acetylation status of the promoter regions of p16^INK4a and p21^WAF1/Cip1 were determined by chromatin immunoprecipitation assay.

Results

A single intravenous injection of FK228 (2.5 mg/ml) successfully inhibited joint swelling, synovial inflammation, and subsequent bone and cartilage destruction in mice with AMA. FK228 treatment induced histone hyperacetylation in the synovial cells and decreased the levels of tumor necrosis factor α and interleukin‐1β in the synovial tissues of mice with AMA. FK228 inhibited the in vitro proliferation of RASFs in a dose‐dependent manner. Treatment of cells with FK228 induced the expression of p16^INK4a and up‐regulated the expression of p21^WAF1/Cip1. These effects of FK228 on p16^INK4a and p21^WAF1/Cip1 were related to the acetylation of the promoter region of the genes.

Conclusion

Our findings strongly suggest that systemic administration of HDA inhibitors may represent a novel therapeutic target in RA by means of cell cycle arrest in RASFs via induction of p16^INK4a expression and increase in p21^WAF1/Cip1 expression.

     

Enhanced age-dependent cerebrovascular dysfunction is mediated by adaptor protein p66

Background

Aging is an independent risk factor for cardiovascular and cerebrovascular disease. To date, little is known about the mechanisms of aging of cerebral arteries and whether the aging gene p66^Shc is implicated in it. The present study was designed to assess age-induced vascular dysfunction in cerebral and systemic arteries of wild type (wt) and p66^Shc −/− mice.

Methods

Basilar arteries and size matched second order femoral arteries of 3-month (3M), 6-month (6M) and 2-year old (2Y) mice were studied in wt and p66^Shc −/− mice. To assess vascular function, arterial rings mounted in a myograph for isometric tension recordings were exposed to increasing concentrations of acetylcholine and sodium nitroprusside. Reactive oxygen species (ROS) generation was assessed in femoral and basilar arteries using the spin trap 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethyl-pyrrolidine.

Results

In wt mice, endothelial function of the femoral artery was not affected by age unlike in the basilar artery where an age-dependent dysfunction was observed. In p66^Shc −/− a similar response was observed in the femoral artery; however, age-dependent endothelial dysfunction of the basilar artery was blunted as compared to wt. Levels of ROS were comparable in the femoral arteries of 3M and 2Y of wt and p66^Shc −/− mice. Differently, ROS levels in the basilar artery of wt mice were strongly increased by age unlike in p66^Shc −/− mice where they remained comparable irrespective of age.

Conclusions

Endothelial function in cerebral arteries, but not in size-matched systemic ones, is heavily impaired by aging. This process is paralleled by an increased ROS production and is mediated by the p66^Shc gene.     

Altered expression of p120catenin predicts poor outcome in invasive breast cancer

Purpose  

The study focuses on p120catenin, a regulator of cell adhesion, which has previously been described in many malignancies and suggested with a role in invasion and metastatic behaviour. In this study, we investigate the role of altered immunoexpression of p120catenin isoforms in the prognosis of invasive breast cancer (n = 351).