High Notch1 protein expression is an early event in breast cancer development and is associated with the HER‐2 molecular subtype

Zardawi S J, Zardawi I, McNeil C M, Millar E K A, McLeod D, Morey A L, Crea P, Murphy N C, Pinese M, Lopez‐Knowles E, Oakes S R, Ormandy C J, Qiu M R, Hamilton A, Spillane A, Soon Lee C, Sutherland R L, Musgrove E A & O’Toole S A
(2010) Histopathology 56, 286–296 High Notch1 protein expression is an early event in breast cancer development and is associated with the HER‐2 molecular subtype Aims: Activation of Notch signalling results in hyperplasia and tumorigenesis in murine mammary epithelium. However, there is little information regarding the expression of Notch1 in premalignant lesions and early breast cancer. We investigated expression of Notch1 in breast cancer development and its association with molecular subtypes. Methods and results: Immunohistochemical expression of Notch1 was determined in a murine model of mammary carcinogenesis and in breast tissue from two cohorts of breast cancer patients, the first (n = 222) comprising a histological progression series and the second an outcome series of 228 patients with operable invasive ductal carcinoma. Enhanced expression of Notch1 protein was an early event in both murine and human breast cancer development with progressive increases in expression with the development of hyperplasia and malignancy. High Notch1 was not prognostic in the outcome cohort. There was, however, a highly significant association of high Notch1 protein with the HER‐2 molecular subtype of breast cancer (P = 0.008). Conclusions: These data demonstrate that aberrant Notch regulation is an early event in mammary carcinogenesis and is associated with the HER‐2 molecular subtype of breast cancer, and suggest the Notch signalling pathway may be a potential therapeutic target worthy of further investigation.     

TUSC3 promotes colorectal cancer progression and epithelial–mesenchymal transition (EMT) through WNT/β‐catenin and MAPK signalling

Colorectal cancer (CRC) is one of the most common malignancies and is the second leading cause of cancer death in humans. Tumour suppressor candidate 3 (TUSC3) plays an important role in embryogenesis and metabolism. Deletion of TUSC3 often causes non‐syndromic mental retardation. Even though TUSC3 deregulation is frequently observed in epithelial cancers, the function of TUSC3 in CRC has remained unknown. In this study, we observed greater expression of TUSC3 at the mRNA and protein level in clinical colorectal tumour samples compared with paired normal tissues. Gain‐ and loss‐of‐function analyses were performed to evaluate the functional significance of TUSC3 in CRC initiation and progression. Immunoblotting, immunofluorescence, and co‐immunoprecipitation analyses were used to identify potential pathways with which TUSC3 might be involved. Overexpression of TUSC3 in CRC cells induced epithelial–mesenchymal transition (EMT) in CRC cells, accompanied by down‐regulation of the epithelial marker, E‐cadherin, and up‐regulation of the mesenchymal marker, vimentin. Increased proliferation, migration, and invasion, as well as accelerated xenograft tumour growth, were observed in TUSC3‐overexpressing CRC cells, while opposite effects were achieved in TUSC3‐silenced cells. In conclusion, our study demonstrated the oncogenic role of TUSC3 in CRC and showed that TUSC3 may be responsible for alternations in the proliferation ability, aggressiveness, and invasive/metastatic potential of CRC through regulating the MAPK, PI3K/Akt, and Wnt/β‐catenin signalling pathways. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

TGFBI protein high expression predicts poor prognosis in colorectal cancer patients

Transforming growth factor-beta-induced (TGFBI) serves as a linker protein and plays a role in the activation of morphogenesis, cell proliferation, adhesion, migration, differentiation and inflammation. High expression levels of the human TGFBI gene are correlated with numerous human malignancies. In order to explore the roles of TGFBI in the tumor progression of colorectal cancer, colorectal cancer specimens from 115 patients with strict follow-up were selected for the analysis of TGFBI by immunohistochemistry. The correlations between TGFBI expression and the clinicopathological features of colorectal cancers were evaluated. In the colorectal cancer tissues, TGFBI was mainly localized in the cytoplasm and stroma and scarcely in the nucleus. TGFBI expression in the cytoplasm and stroma was not found to be associated with age, gender, tumor histopathological grading, PT category and tumor location (P > 0.05 for each). However, high TGFBI expression in the cytoplasm and stroma correlated with lymph node metastasis, distant metastasis and Dukes stage (P < 0.05 for each). The survival rate was significantly lower in patients with high TGFBI expression than in those with low TGFBI expression. Furthermore, we found that tumor node metastasis (TNM) staging (HR: 2.963; 95% CI: 1.573-1.664; P = 0.000), differentiation (HR: 1.574; 95% CI: 1.001-2.476; P = 0.049) and high TGFBI cytoplasmic expression (HR: 3.332; 95% CI: 1.410-7.873; P = 0.000) proved to be independent prognostic factors for survival in colorectal cancer. In conclusion, TGFBI plays an important role in the progression of colorectal cancers and it is an independent poor prognostic factor for colorectal cancer patients.     

Bone morphogenetic protein 4 inhibits liposaccharide‐induced inflammation in the airway

Bone morphogenetic protein 4 (BMP4) is a multifunctional growth factor that belongs to the TGF‐β superfamily. The role of BMP4 in lung diseases is not fully understood. Here, we demonstrate that BMP4 was upregulated in lungs undergoing lipopolysaccharide (LPS)‐induced inflammation, and in airway epithelial cells treated with LPS or TNF‐α. BMP4 mutant (BMP4^+/?) mice presented with more severe lung inflammation in response to LPS or Pseudomonas aeruginosa, and lower bacterial load compared with that in BMP4^+/+ mice. Knockdown of BMP4 by siRNA increased LPS and TNF‐α‐induced IL‐8 expression in 16HBE human airway epithelial cells and in primary human bronchial epithelial cells. Similarly, peritoneal macrophages from BMP4^+/? mice produced greater levels of TNF‐α and keratinocyte chemoattractant (KC) upon LPS treatment compared with cells from BMP4^+/+ mice. Administration of exogenous BMP4 attenuated the upregulation of TNF‐α, IL‐8, or KC induced by LPS and/or TNF‐α in airway epithelial cells, and peritoneal macrophages. Finally, partial deficiency of BMP4 in BMP4^+/? mice protected the animals from restrictive lung function reduction upon chronic LPS exposure. These results indicate that BMP4 plays an important anti‐inflammatory role, controlling the strength and facilitating the resolution of acute lung inflammation; yet, BMP4 also contributes to lung function impairment during chronic lung inflammation.     

Zinc-finger protein X-linked is a novel predictor of prognosis in patients with colorectal cancer

Zinc-finger protein X-linked (ZFX) has been demonstrated to play an important role in the development of human malignancies. However, its prognostic significance in cancer patients remains unclear and less is known about its role in colorectal cancer (CRC). In this study, we found that the expression of ZFX in CRC tissues was significantly higher than that in corresponding normal tissues by quantitative real-time polymerase chain reaction and Western blot. Using immunohistochemistry, we explored the associations between protein expression of ZFX and clinicopathological parameters in 120 CRC cases. The results showed that ZFX expression was significantly associated with tumor differentiation (P = 0.022), tumor size (P = 0.037), tumor invasion (P = 0.027), lymph node metastasis (P = 0.042), distant metastasis (P = 0.011), and Dukes’ classification (P = 0.028). Moreover, according to Kaplan-Meier model, patients with high expression of ZFX had a significantly poorer prognosis compared to those with low expression of ZFX. Multivariate analysis suggested that high expression of ZFX was an independent prognostic factor for CRC patients. In conclusion, our findings for the first time demonstrated that ZFX expression may be associated with the progress of CRC and suggested that ZFX has the potential value to be an effective prognostic predictor for CRC patients.     

Quantitative cell signalling analysis reveals down‐regulation of MAPK pathway activation in colorectal cancer

Mitogen‐activated protein kinases (MAPK) are considered to play significant roles in colonic carcinogenesis and kinase inhibitor therapy has been proposed as a potential tool in the treatment of this disease. Reverse‐phase microarray assays using phospho‐specific antibodies can directly measure levels of phosphorylated protein isoforms. In the current study, samples from 35 cases of untreated colorectal cancer colectomies were laser capture‐microdissected to isolate epithelium and stroma from cancer as well as normal (i.e. uninvolved) mucosa. Lysates generated from these four tissue types were spotted onto reverse‐phase protein microarrays and probed with a panel of antibodies to ERK, p‐ERK, p38, p‐p38, p‐JNK, MEK and p‐MEK. Whereas total protein levels were unchanged, or slightly elevated (p38, p = 0.0025) in cancers, activated isoforms, including p‐ERK, p‐p38 and p‐JNK, were decreased two‐ to four‐fold in cancers compared with uninvolved mucosa (p < 0.0023 in all cases except for p‐JNK in epithelium, where decrement was non‐significant). This was backed up by western blotting. Dukes' stage B and C cancers displayed lower p‐ERK and p‐p38 expression than Dukes' stage A cancers, although this was not statistically significant. It is concluded that MAPK activity may be down‐regulated in colorectal cancer and that further exploration of inhibitory therapy in this system should be carefully evaluated if this finding is confirmed in larger series. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

In Brief: Notch signalling in health and disease

The Notch pathway controls many aspects of development and is mutated in various human cancers. Remarkably, Notch function changes during the development of cellular hierarchies and can be either oncogenic or tumour‐suppressive, depending on cellular context. Notch dysregulation is also important in cardiovascular disease and disorders of immunity. This mini‐review outlines key features of the Notch signalling pathway and emerging data linking it to human diseases. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Bone morphogenetic protein signaling in breast cancer progression

Breast cancer is the most prevalent type of cancer amongst women worldwide. The mortality rate for patients with early-stage breast cancer has been decreasing, however, the 5-year survival rate for patients with metastatic disease remains poor, currently at 27%. Here, we have reviewed the current understanding of the role of bone morphogenetic protein (BMP) signaling in breast cancer progression, and have highlighted the discordant results that are reported in different studies. We propose that some of these contradictory outcomes may result from signaling through either the canonical or non-canonical pathways in different cell lines and tumors, or from different tumor-stromal interactions that occur in vivo.     

肥胖与结直肠癌预后关系的研究进展

结直肠癌(colorectal cancer,CRC)是世界范围内最常见的癌症之一.近年来,随着人们生活方式及饮食结构的改变,肥胖人群的比例越来越多,大量流行病学研究发现,结肠癌的发生风险跟肥胖密切相关.但是,对于肥胖是否影响CRC的预后,研究结果各不相同.文章就肥胖与CRC患者预后的相关性进行阐述,目前临床上对这一问题存在正反两种观点,多数研究结果显示肥胖在很大程度上影响CRC的预后;对肥胖影响CRC预后的可能作用机制进行分析,包括肥胖的胰岛素抵抗/高胰岛素血症、肥胖引起慢性炎症及脂肪酸合成酶(fatty acid synthase,FASN)表达过度.总之,肥胖作为CRC的主要危险因素及不良预后因素,在一定程度上影响CRC病死及复发.     

Systematic assessment of protein phenotypes characterizing high‐grade tumour budding in mismatch repair‐proficient colorectal cancer

Karamitopoulou E, Lugli A, Panayiotides I, Karakitsos P, Peros G, Rallis G, Patsouris E S, Terracciano L & Zlobec I (2010) Histopathology 57 , 233–243
Systematic assessment of protein phenotypes characterizing high‐grade tumour budding in mismatch repair‐proficient colorectal cancer Aims: A tumour bud is defined as a single tumour cell or tumour cell cluster of up to five cells at the invasive tumour front. Significant differences in survival have been detected in colorectal cancer patients with low‐ compared to high‐grade budding. The aim of this study was to identify potential multi‐marker phenotypes characterizing low‐ and high‐grade budding in mismatch repair (MMR)‐proficient colorectal cancer. Methods and results: Established and promising prognostic proteins such as epidermal growth factor receptor (EGFR), pERK, RHAMM, RKIP, β‐catenin, E‐cadherin, pAKT, p16, p21, Ki67, Bcl‐2, vascular endothelial growth factor (VEGF), apoptotic protease activating factor‐1 (APAF‐1), MUC1, EphB2, matrix metalloproteinase 7, pSMAD2, CDX2, laminin5γ2 and MST1 were analysed on 208 MMR‐proficient colorectal cancers with complete clinicopathological data. The most accurate markers for predicting high‐grade budding (more than six tumour buds) were EphB2 (P < 0.001), Bcl‐2 (P < 0.001), RKIP (P < 0.001), E‐cadherin (P = 0.004), laminin5γ2 (P = 0.004) and APAF‐1 (P = 0.005). On multivariable analysis, only loss of Bcl‐2 (P < 0.001) and EphB2 (P < 0.001) were independent predictors of high‐grade budding. Bcl‐2?/EphB2? tumours were more frequently poorly differentiated (P < 0.001), of advanced pT stage (P = 0.002), lymph node positive (P = 0.023), presented vascular (P = 0.053) and lymphatic invasion (P = 0.005) and had a negative impact on patient survival (P = 0.012). Conclusions: The multi‐marker phenotype EphB2?/Bcl‐2? is an independent predictor of high‐grade budding and implies increased aggressive behaviour in MMR‐proficient colorectal cancer.     

Association between KIAA1199 overexpression and tumor invasion,TNM stage,and poor prognosis in colorectal cancer

To investigate the expression of KIAA1199 in tumor tissue and its potential value as a prognostic indicator of survival in patients with colorectal cancer (CRC). The expression of KIAA1199 mRNA in CRC was characterized using real-time PCR and 20 pairs of fresh-frozen CRC tissues and corresponding non-cancerous tissues. KIAA1199 protein expression was confirmed using immunohistochemistry on a tissue microarray chip from 202 patients with CRC. Then, we correlated KIAA1199 protein expression to CRC conventional clinicopathological features and patient’s outcome. The expression of KIAA1199 mRNA and protein were up-regulated in CRC compared to normal tissues (P = 0.015 and P < 0.001, individually). KIAA1199 protein expression was related to tumor invasion depth (P = 0.013) and lymph node metastasis (P = 0.003). Kaplan-Meier survival and Cox regression analyses revealed that high KIAA1199 expression (P < 0.001) and serum carcinoembryonic antigen (CEA) level post operation (P = 0.005) were independent factors predicting poor prognosis of patients with CRC. We present evidence that high expression of KIAA1199 is associated with tumor invasion depth, TNM stage, and poor prognosis in CRC. Our findings suggest KIAA1199 could be used as a prognostic factor and novel therapeutic target for CRC.     

Expression of HER2 in colorectal cancer does not correlate with prognosis

Estimation of HER2 membranous expression is routinely used in breast and gastric cancers, as both a prognostic and a predictive factor. To date there is no evidence for similar application of HER2 expression in colorectal cancer (CRC) cells. In CRC, HER2 is sometimes overexpressed in the cell membrane and very often in the cytoplasm. This study was conducted to determine possible correlations between both membranous and cytoplasmatic expression of HER2 in CRC cells and the outcome of the disease. The prognostic significance of combined staining intensity in the cell membrane and cytoplasm in the entire CRC cell was also investigated. HER2 expression in resectable colorectal adenocarcinoma cells was evaluated by immunohistochemistry in specimens taken from 202 patients. The percentage of cancer cells with membranous or cytoplasmatic reactions and the staining intensity of the reaction in the whole cell were recorded. A membranous reaction was present in 27% of cases, and cytoplasmatic reaction in 66% of cases. The total staining intensity in the entire cell was evaluated as moderate (2+) in 32% of cases and strong (3+) staining in 15%. There was no correlation found between either membranous or cytoplasmatic HER2 expression and survival. Furthermore combined staining intensity did not provide any prognostic information. We conclude that HER2 expression in CRC does not correlate with prognosis.     

A pyroptosis-related prognosis model to predict survival in colorectal cancer patients

Pyroptosis is a recently-identified pathway of host cell death that is stimulated by a range of microbial infections. Emerging evidence indicates pyroptosis plays crucial roles in tumor growth, disease progression, and migration of different cancer cells. However, the clinical significance of pyroptosis in tumor behavior prognosis, as well as the underlying mechanism in different cancers remains elusive. Here, by evaluating the expression level of pyroptosis genes in colorectal cancer (CRC) patients from the TCGA cohort and GEO cohort ({"type":"entrez-geo","attrs":{"text":"GSE39582","term_id":"39582"}}GSE39582), we identified pyroptosis-related DEGs and then built a 13-gene risk model by applying the LASSO Cox regression algorithm. Furthermore, functional analysis using GSEA and GSEV revealed that our prognostic model may function through regulating immune responses and tumor biogenesis pathways. Significant infiltration of activated immune cells (e.g. cytotoxic T cells) was observed in the low risk score group. The selected gene set was further validated in the GEO cohort. Time-dependent ROC curves confirmed that our risk score model is robust in predicting 1, 3 and 5-year overall survival in CRC patients. Overall, we have identified a pyroptosis-related gene signature that consists of 13 genes, which serves as a potent indicator of CRC prognosis. Thus, our model provides insights in how to make better clinical decision in the future.     

Somatic frameshift mutations of bone morphogenic protein receptor 2 gene in gastric and colorectal cancers with microsatellite instability

Mounting evidence exists that perturbation of bone morphogenic protein (BMP) signaling is involved in cancer development, especially in gastrointestinal cancers. However, somatic mutations of the genes encoding BMP and BMP receptors have not yet been discovered in human cancer tissues. By analyzing a public database, we found that BMP receptor 2 (BMPR2) and BMP1 genes had mononucleotide repeats in their coding sequences that could be mutation targets in cancers with microsatellite instability (MSI). In this study, we analyzed the mutation of BMPR2 and BMP1 genes in gastric (GC) and colorectal cancers (CRC) with MSI [31 GC with high MSI (MSI-H), 13 GC with low MSI (MSI-L), 38 CRC with MSI-H and 15 CRC with MSI-L] by single-strand conformation polymorphism analysis and DNA sequencing. Overall, we found seven frameshift mutations in the BMPR2 gene, but not in the BMP1 gene. The mutations were an identical deletion mutation of one base in the repeats (c.1748delA) that would result in premature stops of the amino acid synthesis (p.Asn583ThrfsX44). The BMPR2 mutations were detected in 6.5% of GC and 13.2% of CRC with MSI-H. All the cancers with the BMPR2 mutation showed loss of BMPR2 expression. Our data indicate that frameshift mutation of BMPR2 gene occurs in GC and CRC with MSI-H, and suggest that the BMPR2 mutation might contribute to cancer pathogenesis by inactivating BMPR2-mediated BMP signaling.