The article presents the results of a study of the dynamics and structure of the incidence of a new coronavirus infection (COVID-19) in the Orenburg region for 2020–2022, in view of the circulating genovariant of the SARS-CoV-2 virus. The data of operational monitoring of the Department of Rospotrebnadzor used in the Orenburg region. On the territory of the Orenburg region, the circulating genovariant of the coronavirus infection and its influence on the incidence rate, age, sex of patients and the clinical form of diseases, 3 periods allocated: The 1st period (March 2020 — March 2021) characterized by the circulation of the Wuhan SARS-CoV-2 genovariant; in the 2nd period (April — December 2021), the SARS-CoV-2 Delta genovariant and, along with it, the British SARS-CoV-2 Alpha genovariant became widespread; in 3In the 1st period (January-December 2022), the SARS-CoV-2 Omicron gene variant prevailed. The incidence rate in every subsequent period was significantly higher than in the previous one. In all periods, women have highest morbidity, the age group at risk was people over 65 years old. The clinical variants of the disease was heterogeneous: the dynamics showed an increase in the proportion of acute respiratory viral infections, a decrease in the proportion of pneumonia and asymptomatic forms. The most severe course of the disease and the highest mortality established in the second period, against the background of the intensive dissemination of the most virulent Delta genvariant. The circulation of the contagious Omicron genevariant can be explained the peak values of morbidity at the beginning of the third period. The territorial unevenness of the distribution of morbidity revealed. The correlation established between the circulating genovariants of the SARS-CoV-2 virus and the manifestations of the epidemic process of a new coronavirus infection (COVID-19) in the Orenburg Region in 2020–2022.

Supplementary Table 2 from Notch Signaling Activated by Replication Stress–Induced Expression of Midkine Drives Epithelial–Mesenchymal Transition and Chemoresistance in Pancreatic Cancer

Supplementary Table 1 from Notch Signaling Activated by Replication Stress–Induced Expression of Midkine Drives Epithelial–Mesenchymal Transition and Chemoresistance in Pancreatic Cancer

Supplementary Figure Legends 1-5 from Notch Signaling Activated by Replication Stress–Induced Expression of Midkine Drives Epithelial–Mesenchymal Transition and Chemoresistance in Pancreatic Cancer

Cell migration is one of the hallmarks of metastatic disease and thus identification of migration promoting proteins is crucial for the understanding of metastasis formation. Here we show that the neuron-specific, F-actin bundling inositol-1,4,5-trisphosphate-3-kinase-A (ITPKA) is ectopically expressed in tumor cells and critically involved in migration. Down-regulation of ITPKA expression in transformed cell-lines with ectopic expression of ITPKA significantly decreased migration and the number of linear and branched cell protrusion. Conversely, up-regulation of ITPKA in tumor cell lines with low endogenous ITPKA expression increased migration and formation of cell processes. In vitro, ITPKA alone induced the formation of linear actin filaments, whereas ITPKA mediated formation of branched protrusions seems to result from interaction between ITPKA and the F-actin cross-linking protein filamin C. Based on these actin-modulating and migration-promoting effects of ITPKA we examined its expression in clinical samples of different tumor entities, starting with the analysis of multiple tumor tissue arrays. As in lung adenocarcinoma specimens, the highest ITPKA expression rate was found, this tumor entity was examined in more detail. ITPKA was expressed early in adenocarcinoma progression (pN0) and was largely maintained in invasive and metastatic tumor cell populations (pN1/2, lymph node metastases). Together with our result that high expression of ITPKA increases motility of tumor cells we conclude that the observed expression of ITPKA early in tumor development increases the metastatic potential of lung adenocarcinoma cells. Therefore, we suggest that ITPKA may be a promising therapeutic molecular target for anti metastatic therapy of lung cancer.

Abstract The incidence of pancreatic ductal adenocarcinoma (PDAC) nearly equals its mortality rate, partly because most PDACs are intrinsically chemoresistant and thus largely untreatable. It was found recently that chemoresistant PDAC cells overexpress the Notch-2 receptor and have undergone epithelial–mesenchymal transition (EMT). In this study, we show that these two phenotypes are interrelated by expression of Midkine (MK), a heparin-binding growth factor that is widely overexpressed in chemoresistant PDAC. Gemcitabine, the front-line chemotherapy used in PDAC treatment, induced MK expression in a dose-dependent manner, and its RNAi-mediated depletion was associated with sensitization to gemcitabine treatment. We identified an interaction between the Notch-2 receptor and MK in PDAC cells. MK–Notch-2 interaction activated Notch signaling, induced EMT, upregulated NF-κB, and increased chemoresistance. Taken together, our findings define an important pathway of chemoresistance in PDAC and suggest novel strategies for its clinical attack. Cancer Res; 71(14); 5009–19. ©2011 AACR.

Insulin‐like growth factor‐1 receptor (IGF‐1R) and human epidermal growth factor receptor‐2 (HER2) receptor expression has been found to be a key regulator of tumorigenesis. The purpose of our study was to establish the prognostic significance of IGF‐1R in esophageal cancer and to determine the effect of IGF‐1R and HER2 targeting with α‐IR3 and Herceptin™ antibodies on the proliferation of esophageal cancer cells in vitro. IGF‐1R expression and clinicopathological correlations were analyzed with a tissue microarray containing 234 esophageal cancer specimens (133 adenocarcinomas and 101 squamous cell carcinomas). Proliferation changes associated with Herceptin™ and α‐IR3 blockage were evaluated with the unique human esophageal cancer cell lines Pt1590 and LN1590. IGF‐1R and HER2 expression levels, activation and phosphorylation status of downstream signaling proteins involved in the activation pathways were analyzed by Western blotting. IGF‐1R overexpression was detected in 121 (52%) of the 234 esophageal tumors examined. In the subgroup of 87 HER2‐positive tumors, 93.1% showed concordant overexpression for IGF‐1R. IGF‐1R was identified as a variable associated with reduced overall survival for adenocarcinoma (p = 0.05), but not for squamous cell carcinoma. The combination of Herceptin™ and α‐IR3 was more effective in inhibiting in vitro proliferation than treatment with either agent alone (p < 0.01). This was associated with a decrease in HER2 and IGF‐1R protein levels and suppression of Akt‐ and MAP kinase phosphorylation. IGF‐1R expression can be used as a novel prognostic marker for adenocarcinomas of the esophagus. Cotreatment with IGF‐1R and HER2 antibodies might become a valuable and effective treatment option in esophageal adenocarcinoma.

We describe the development of an aggressive orthotopic metastatic model of esophageal cancer, which is visualized in real time with combined magnetic resonance imaging (MRI) and fluorescence imaging. The aim of the study was to describe the development of a novel model of metastatic tumor disease of esophageal carcinoma and use this model to evaluate fluorescence and MRI in early detection of local and metastatic disease. The human esophageal adenocarcinoma cell line PT1590 was stably transfected with green fluorescent protein (GFP). Nude mice were orthotopically implanted with PT1590-GFP cells. Orthotopic tumor growth as well as metastatic spread was examined by fluorescence imaging and high-resolution MRI at defined intervals after orthotopic implantation. Highly aggressive novel fluorescent cell lines were isolated from metastatic tissues and put into culture. After implantation of these cells, 100% of the animals developed orthotopic primary tumors. In 83% of animals, metastatic spread to liver, lung and lymph nodes was observed. Primary tumor growth could be visualized with fluorescence imaging and with MRI with high correlation between the 2 methods. Fluorescence imaging allows fast, sensitive, and economical imaging of the primary and metastatic tumor without anesthesia. With MRI, anatomical structures are visualized more precisely and tumors can be more accurately localized to specific organs. This model should prove highly useful to understand esophageal carcinoma and to identify novel therapeutics for this treatment-resistant disease.

BACKGROUND:Pancreatic cancer is still associated with devastating prognosis. Real progress in treatment options has still not been achieved. Therefore new models are urgently needed to investigate this deadly disease. As a part of this process we have established and characterized a new human pancreatic cancer cell line.METHODS:The newly established pancreatic cancer cell line PaCa 5061 was characterized for its morphology, growth rate, chromosomal analysis and mutational analysis of the K-ras, EGFR and p53 genes. Gene-amplification and RNA expression profiles were obtained using an Affymetrix microarray, and overexpression was validated by IHC analysis. Tumorigenicity and spontaneous metastasis formation of PaCa 5061 cells were analyzed in pfp-/-/rag2-/- mice. Sensitivity towards chemotherapy was analysed by MTT assay.RESULTS:PaCa 5061 cells grew as an adhering monolayer with a doubling time ranging from 30 to 48 hours. M-FISH analyses showed a hypertriploid complex karyotype with multiple numerical and unbalanced structural aberrations. Numerous genes were overexpressed, some of which have previously been implicated in pancreatic adenocarcinoma (GATA6, IGFBP3, IGFBP6), while others were detected for the first time (MEMO1, RIOK3). Specifically highly overexpressed genes (fold change > 10) were identified as EGFR, MUC4, CEACAM1, CEACAM5 and CEACAM6. Subcutaneous transplantation of PaCa 5061 into pfp-/-/rag2-/- mice resulted in formation of primary tumors and spontaneous lung metastasis.CONCLUSION:The established PaCa 5061 cell line and its injection into pfp-/-/rag2-/- mice can be used as a new model for studying various aspects of the biology of human pancreatic cancer and potential treatment approaches for the disease.

Abstract Purpose: Pancreatic cancer (PC) is a deadly disease. Chemotherapy options are limited to gemcitabine and 5-fluorouracil (5-FU). The molecular profile of PC offers several growth receptors (epidermal growth factor receptor [EGFR], insulin-like growth factor 1 receptor [IGF-1R]) and tyrosine kinases (AKT and MAPK) as promising therapy targets. EGFR, IGF-1R, AKT and MAPK are dependent upon the heat shock protein 90 (HSP90) to gain the mature functional active protein form hence HSP90 represents a promising therapy target in PC. Functional inhibition of HSP90 has the advantage of disruption of many oncogenic signalling pathways simultaneously. The aim of this study was evaluate the efficacy of different HSP90 inhibitors in gemcitabine and 5-FU resistant PC. Material and Methods: Human gemcitabine and 5-FU resistant pancreatic cancer cell lines 5061, 5072 and 5156 were generated and brought in to culture from patients who were operated for PC at our institution and received gemcitabine as adjuvant therapy. The commercial L3.6pl cell line served as a control. Antiproliferative efficacy of three different HSP90 inhibitors (17AAG, 17DMAG and 17AEPGA) was evaluated by the MTT assay. Alterations in signalling pathway effectors and apoptosis upon HSP90 inhibition was determined by western blot analysis (WB). Results: The cell lines 5061, 5072 and 5156 were resistant to gemcitabine and 5-FU. In contrast, 17AAG and the water-soluble derivates 17DMAG and 17AEPGA displayed high antiproliferative activity in all tested cell lines. The calculated GI50 was below 1 µM. In line with the proliferation inhibition in the MTT assay, the expression of EGFR, IGF-1R, AKT and MAPK was significantly down regulated under HSP90 inhibition as verified inWB. In contrast, HSP70 and HSP27 were upregulated in all cell lines under HSP90 inhibition. Apoptosis was an early event in all three HSP90 inhibited cell lines and determined by CASPASE-3 and poly (ADP-ribose) polymerase assay. In contrast, gemcitabine treated cells did not show significant apoptosis. Conclusions: Functional inhibition of HSP90 disrupts multiple signalling cascades in gemcitabine and 5-FU resistant PC. Water-soluble 17DMAG is equally effective as 17AAG. The up-regulation of HSP70 and HSP27 indicates counter regulation of tumor cells to prevent apoptosis and should be addressed when considering HSP90 inhibition in PC. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):B86.