Prognostic significance of HER3 and HER4 protein expression in colorectal adenocarcinomas

BACKGROUND: Colorectal cancer remains a major cause of cancer mortality in the Western world. A limited number of studies has been conducted in respect of Her-3 and Her-4 expression and their correlation with clinical parameters and prognosis in colorectal carcinomas . In this study we sought to determine the pattern and the prognostic significance of HER-3 and HER-4 in colorectal adenocarcinoma. METHODS: We studied HER-3 and HER-4 protein expression in106 paraffin embedded specimens of primary colorectal tumors using immunohistochemistry. The pattern and protein expression levels of HER-3 and HER-4 were correlated with several clinical and pathological parameters. RESULTS: HER-3 staining displayed membranous and cytoplasmic expression pattern in 18 (17%) and 30 samples (28,3%), respectively. HER-4 membranous and cytoplasmic expression was found in 20 (18,9%) and 32 samples (30,2%), respectively. Specimens regarded as positive for HER-3 cytoplasmic expression were associated with moderate tumor grade (p = 0,032) and older median age (p = 0,010). Specimens regarded as positive for HER-4 membranous protein expression were associated with involved lymphnodes (p = 0,0003). Similar results were obtained when considering Her-3 and Her-4 protein expression irrespective of their cellular localization. There was no correlation between the expression of HER-3 and HER-4 and patients outcome. CONCLUSION: HER-4 membranous protein expression was found to predict for lymph nodes positivity in this cohort of patients with colorectal cancer.HER-4 expression status may identify tumors with aggressive biological behavior and increased metastatic potential

Cetuximab potentiates oxaliplatin cytotoxic effect through a defect in NER and DNA replication initiation

Preclinical studies have demonstrated that the chemotherapeutic action of oxaliplatin, a third generation platinum derivative, is improved when combined with cetuximab, a monoclonal antibody inhibitor of epidermal growth factor receptors. To explore the mechanism of this synergistic benefit, we used HCT-8 and HCT-116, two human colon cancer cell lines, respectively, responsive and non-responsive to the oxaliplatin/cetuximab combination. We examined the effect of drug exposure on glutathione-S-transferase-mediated oxaliplatin detoxification, DNA–platinum adducts formation, cell cycle distribution, apoptosis, and the expression of multiple targets involved in DNA replication, recombination, and repair. The major changes we found in HCT-8 were a stimulation of oxaliplatin–DNA adduct formation associated with reduced expression of the key enzyme (excision repair cross complementation group1: ERCC1) in the key repair process of oxaliplatin–DNA platinum adduct, the nucleotide excision repair (NER), both at the mRNA and protein levels. We also observed a reduced expression of factors involved in DNA replication initiation, which correlates with an enrichment of cells in the G1 phase of the cell cycle as well as an acceleration of apoptosis. None of these changes occurred in the non-responsive HCT-116 cell that we used as a negative control. These findings support the fact that cetuximab potentiates the oxaliplatin-mediated cytotoxic effect as the result of inhibition of NER and also DNA replication initiation

Phase II study of erlotinib (OSI-774) in patients with metastatic colorectal cancer

Erlotinib (Tarceva™, OSI-774), a potent epidermal growth factor receptor tyrosine kinase inhibitor (EGFR), was evaluated in a phase II study to assess its activity in patients with metastatic colorectal cancer. In all, 38 patients with metastatic colorectal cancer were treated with erlotinib at a continuous daily oral dose of 150 mg. Radiological evaluation was carried out every 8 weeks and tumour biopsies were performed before treatment and on day 8. Of 31 evaluable patients, 19 (61%) had progressive disease and 12 (39%) had stable disease (s.d.). The median time to progression for those patients having s.d. was 123 days (range 108–329 days). The most common adverse events were rash in 34 patients and diarrhoea in 23 patients. Correlative studies were conducted to investigate the effect of erlotinib on downstream signalling. Tumour tissue correlations were based on usable tissue from eight match paired tumour samples pre- and on therapy, and showed a statistically significant decrease in the median intensity of both pEGFR (P=0.008) and phospho-extracellular signal-regulated kinase (ERK) (P=0.008) a week after commencement of treatment. No other statistically significant change in tumour markers was observed. Erlotinib was well tolerated with the most common toxicities being rash and diarrhoea. More than one-third of evaluable patients had s.d. for a minimum of 8 weeks. Correlative studies showed a reduction in phosphorylated EGFR and ERK in tumour tissue post-treatment

Targeting EGFR and HER-2 with cetuximab- and trastuzumab-mediated immunotherapy in oesophageal squamous cell carcinoma

We previously reported that oesophageal squamous cell carcinoma (SCC) had a relatively high incidence of EGFR and HER-2 overexpression. Thus, anti-HER family targeting may become a promising approach to treat oesophageal SCC. In the present study, we investigated (a) the distribution of EGFR and HER-2 expression in oesophageal SCC (n=66) detected by immunohistochemistry and (b) cetuximab- and/or trastuzumab-mediated biological activity (antiproliferative effect by the MTT assay, apoptosis-inducing activity by the annexin V/propidium iodide assay, and antibody-dependent cellular cytotoxicity (ADCC) by the 51Cr-release assay) against oesophageal SCC cell lines with various levels of EGFR and HER-2. Twelve of the 66 patients (18%) showed both EGFR- and HER-2 expression. Out of both EGFR- and HER-2-positive cases, nine cases (75%) showed EGFR and HER-2 expression in individually distinct regions. Furthermore, the combination of cetuximab and trastuzumab could induce synergistic antiproliferative effects and additional ADCC activities against not all, but several oesophageal SCC cell lines with EGFR and HER-2 expression. The combination of cetuximab and trastuzumab may be useful in the treatment of oesophageal SCC with EGFR and HER-2 expression

From proteomic analysis to potential therapeutic targets: functional profile of two lung cancer cell lines, A549 and SW900, widely studied in pre-clinical research

Lung cancer is a serious health problem and the leading cause of cancer death worldwide. The standard use of cell lines as in vitro pre-clinical models to study the molecular mechanisms that drive tumorigenesis and access drug sensitivity/effectiveness is of undisputable importance. Label-free mass spectrometry and bioinformatics were employed to study the proteomic profiles of two representative lung cancer cell lines and to unravel the specific biological processes. Adenocarcinoma A549 cells were enriched in proteins related to cellular respiration, ubiquitination, apoptosis and response to drug/hypoxia/oxidative stress. In turn, squamous carcinoma SW900 cells were enriched in proteins related to translation, apoptosis, response to inorganic/organic substances and cytoskeleton organization. Several proteins with differential expression were related to cancer transformation, tumor resistance, proliferation, migration, invasion and metastasis. Combined analysis of proteome and interactome data highlighted key proteins and suggested that adenocarcinoma might be more prone to PI3K/Akt/mTOR and topoisomerase IIα inhibitors, and squamous carcinoma to Ck2 inhibitors. Moreover, ILF3 overexpression in adenocarcinoma, and PCNA and NEDD8 in squamous carcinoma shows them as promising candidates for therapeutic purposes. This study highlights the functional proteomic differences of two main subtypes of lung cancer models and hints several targeted therapies that might assist in this type of cancer.publishe

Reciprocal priming between receptor tyrosine kinases at recycling endosomes orchestrates cellular signalling outputs

From Wiley via Jisc Publications RouterHistory: received 2020-10-29, rev-recd 2021-04-27, accepted 2021-04-28, pub-electronic 2021-06-04Article version: VoRPublication status: PublishedFunder: Wellcome Trust; Grant(s): 107636/Z/15/Z, 210002/Z/17/ZFunder: UKRI | Biotechnology and Biological Sciences Research Council (BBSRC); Id: http://dx.doi.org/10.13039/501100000268; Grant(s): BB/R015864/1, BB/M011208/1Funder: UKRI | Medical Research Council (MRC); Id: http://dx.doi.org/10.13039/501100000265; Grant(s): MR/T016043/1Funder: Cancer Research UK (CRUK); Id: http://dx.doi.org/10.13039/501100000289; Grant(s): A27445Funder: NIHR Manchester Biomedical Research Centre; Grant(s): IS‐BRC‐1215‐20007Funder: Breast Cancer Now; Grant(s): MAN‐Q2‐Y4/5Abstract: Integration of signalling downstream of individual receptor tyrosine kinases (RTKs) is crucial to fine‐tune cellular homeostasis during development and in pathological conditions, including breast cancer. However, how signalling integration is regulated and whether the endocytic fate of single receptors controls such signalling integration remains poorly elucidated. Combining quantitative phosphoproteomics and targeted assays, we generated a detailed picture of recycling‐dependent fibroblast growth factor (FGF) signalling in breast cancer cells, with a focus on distinct FGF receptors (FGFRs). We discovered reciprocal priming between FGFRs and epidermal growth factor (EGF) receptor (EGFR) that is coordinated at recycling endosomes. FGFR recycling ligands induce EGFR phosphorylation on threonine 693. This phosphorylation event alters both FGFR and EGFR trafficking and primes FGFR‐mediated proliferation but not cell invasion. In turn, FGFR signalling primes EGF‐mediated outputs via EGFR threonine 693 phosphorylation. This reciprocal priming between distinct families of RTKs from recycling endosomes exemplifies a novel signalling integration hub where recycling endosomes orchestrate cellular behaviour. Therefore, targeting reciprocal priming over individual receptors may improve personalized therapies in breast and other cancers

Effect of supercritical carbon dioxide on inactivation and germination of alicyclobacillus acidoterrestris spores

Przetrwalniki termofilnych kwasolubnych bakterii Alicyclobacillus acidoterrestris są oporne na działanie czynników zewnętrznych i mogą się rozwijać w sokach w czasie przechowywania. Wytwarzają związki o dezynfekcyjnym zapachu, m. in. gwajakol i prowadzą do zepsucia soków. Zwiększenie stopnia kiełkowania przetrwalników w procesie utrwalania żywności może przyczynić się do ich skutecznej inaktywacji. Celem pracy była charakterystyka przebiegu procesu kiełkowania i stopnia inaktywacji przetrwalników Alicyclobacillus acidoterrestris zainicjowanego przez ditlenek węgla w stanie nadkrytycznym (SCCD). Przetrwalniki zawieszone w soku jabłkowym oraz w buforach o pH 4,0 i pH 7,0 poddawano działaniu SCCD o ciśnieniu 10 ÷ 60 MPa, w temp. 50 ÷ 75 ºC, w ciągu 20 ÷ 40 min. Po 40 min działania SCCD o ciśnieniu 60 MPa w temp. 75 ºC liczba skiełkowanych przetrwalników w soku jabłkowym wynosiła 3,9 log, z czego 3,4 log było inaktywowanych. W tych samych warunkach procesu, ale w buforze o pH 4,0 uzyskano niższy stopień kiełkowania (3,2 log) i inaktywacji (2,7 log). W buforze o pH 7,0 stwierdzono stopień wykiełkowania rzędu 2,5 log i inaktywację przetrwalników o 1,1 log. Stwierdzono istotną korelację pomiędzy wynikami szacowania stopnia kiełkowania uzyskanymi metodą płytkową i metodą pomiaru zmniejszania gęstości optycznej. Temperatura stosowanego SCCD okazała się czynnikiem istotnie wpływającym na stopień kiełkowania i inaktywację przetrwalników A. acidoterrestris. Wraz z obniżaniem pH zaobserwowano niewielkie zwiększenie stopnia kiełkowania i znaczne zwiększenie inaktywacji przetrwalników A. acidoterrestris poddanych działaniu SCCD, a zawartość składników odżywczych w soku jabłkowym dodatkowo zwiększała stopień wykiełkowania.The spores of thermophilic, acidophilic bacterium Alicyclobacillus acidoterrestris are resistant to external factors and can develop in juices during storage. They produce compounds that smell like disinfectants, i.a., guaiacol, and lead to the spoilage of juices. The increasing of the degree of spore germination during the process of food preservation can contribute to their effective inactivation. The objective of the research study was to characterise the course of the germination process and the inactivation degree of Alicyclobacillus acidoterrestris spores initiated by supercritical carbon dioxide (SCCD). The spores suspended in apple juice and in buffers of pH 4.0 and pH 7.0 were treated with SCCD at a pressure of 10-60 MPa, at temperatures of 50 ÷ 75 ºC, for 20 ÷ 40 min. After 40 min SCCD treatment at 60 MPa and at 75 ºC, the count of spore germination in apple juice was 3.9 log, of which 3.4 log were inactivated. In the pH 4.0 buffer and under the same process conditions, a lower degree of germination (3.2 log) and inactivation (2.7log) was achieved. In the pH 7.0 buffer, the germination of 2.5 log was reported and the spore inactivation of 1.1 log. A significant correlation was found between the results of germination degree estimation that were obtained using a plate method and a method of measuring the decrease in optical density. The temperature of the SCCD applied turned out to be a factor to significantly impact the degree of germination and the inactivation of A. acidoterrestris spores. Along with the decrease in pH, a slight increase in the degree of spore germination was observed and a significant increase in the inactivation of the A. acidoterrestris spores treated with SCCD; the content of nutrients in apple juice further increased the degree of germination

Nutritional knowlegde and lifestyle of obese patients with diagnosed obturative sleep apnea

Celem pracy była ocena wiedzy żywieniowej i i wybranych elementów stylu życia pacjentów otyłych z rozpoznanym OBS, leczonych w Dolnośląskim Centrum Chorób Płuc Akademii Medycznej we Wrocławiu. Ocenę stanu wiedzy żywieniowej przeprowadzono wśród 49 chorych, w tym 12 kobiet i 37 mężczyzn z rozpoznanym OBS, leczonych w Dolnośląskim Centrum Chorób Płuc we Wrocławiu. Do oceny wiedzy żywieniowej i wybranych elementów stylu życia wykorzystano autorski, standaryzowany kwestionariusz opracowany w Zakładzie Żywienia Człowieka Uniwersytetu Przyrodniczego we Wrocławiu. Zawierał on różne pytania, dotyczące m.in. rodzaju aktywności fizycznej, częstości palenia papierosów. Mimo wykazanej częściowo dobrej wiedzy żywieniowej, jej wykorzystanie przez chorych na OBS w życiu codziennym budzi wiele zastrzeżeń. Może to prawdopodobnie wynikać z nie wiązaniem wiadomości o prawidłowym żywieniu z własnym stanem zdrowia. Stwierdzona w badanej grupie chorych na OBS otyłość i niska aktywność fizyczna, świadczą o konieczności włączenia do procesu leczenia indywidualnego poradnictwa dietetycznego.The study was aimed, therefore, at evaluating the nutritional knowledge and selected elements of lifestyle of obese patients with diagnosed obstructive sleep apnea (OSA), treated at the Lower Silesia Center of Pulmonary Diseases in Wroclaw. Assessment of nutritional knowledge was conducted among 49 patients including 12 women and 37 men with diagnosed obstructive sleep apnea (OSA), treated at the Lower Silesia Center of Pulmonary Diseases in Wroclaw. The nutritional knowledge and selected elements of lifestyle were evaluated by means of own-construct, standardized questionnaire elaborated at the Chair of Human Nutrition, Wroclaw University of Life and Environmental Sciences. It included questions from different fields referring to, among others: knowledge on rational nutrition, type of physical activity, frequency of cigarette smoking, etc. Despite the demonstrated some knowledge of good nutrition, its use by patients with OSA in everyday life raises many objections. This could possibly result from the message is not binding on the correct nutrition to their own health. Found in the studied group of patients with OSA obesity and low physical activity, evidence of the need to integrate into the process of treatment of individual dietary counseling