KRAS mutation analysis in ovarian samples using a high sensitivity biochip assay

<p>Abstract</p> <p>Background</p> <p>Mutations in the <it>KRAS </it>gene are one of the most frequent genetic abnormalities in ovarian carcinoma. They are of renewed interest as new epidermal growth factor receptor (EGFR)-targeted therapies are being investigated for use in ovarian carcinoma. As <it>KRAS </it>mutations are associated with poor response and resistance to EGFR-targeting drugs, this study was conducted to obtain more information on the spectrum of <it>KRAS </it>mutations in ovarian carcinoma.</p> <p>Methods</p> <p>The presence of <it>KRAS </it>mutations in codon 12 and 13 was analyzed in frozen and formalin-fixed paraffin-embedded (FFPE) tissue with a low density biochip platform. 381 malignant (29 borderline malignancy, 270 primary carcinomas, and 82 recurrent carcinomas) and 22 benign tissue samples from a total of 394 patients were examined. <it>KRAS </it>mutational status of each sample was correlated with dignity, FIGO stage, grade, histology, and survival.</p> <p>Results</p> <p><it>KRAS </it>mutations were found in 60 (15%) samples with 58 samples deriving from malignant tissue and 2 samples deriving from benign tissue. In 55 (92%) samples codon 12 was found to be mutated. Frozen and FFPE samples concurred with respect to <it>KRAS </it>mutation status.</p> <p>Conclusion</p> <p><it>KRAS </it>mutation is a common event in ovarian cancer primarily in carcinomas of lower grade, lower FIGO stage, and mucinous histotype. The <it>KRAS </it>mutational status is no prognostic factor for patients treated with standard therapy. However, in line with experience from colorectal cancer and non-small-cell-lung cancer (NSCLC), it may be important for prediction of response to EGFR-targeted therapies.</p

Results of a German wide survey towards current surgical approach in early stage cervical cancer NOGGO MONITOR 11

Minimally invasive surgery (MIS) has become the standard approach in early stage cervical cancer (ECC). However, the recently published "LACC" trial and even others could show inferior PFS and OS of MIS compared to open radical hysterectomy. The results led to a widespread debate about the best surgical approach in ECC. The present survey aimed to get first insights after publication. NOGGO and AGE conducted a nationwide digital survey among 186 Gynecological Cancer Centers. Descriptive statistics and t-tests were performed using SPSS. A majority of the centers were of high expertise and/or experience in treatment of ECC and were highly aware of the LACC trial results. Trial quality and scientific value were rated as very good/good. However, still 40% would not change the standard of care to open surgery. Centers with higher volume and participating in clinical trials were more likely to change. This survey represents insights after the surprising results of recently published trials towards the surgical approach of ECC. There still seems to be a high need of future trials and possible explanations for the unexpected worse outcomes in the MIS group

Variants of the branches of the coeliac trunk

Catedra Anatomia OmuluiThis article represents a study of the coeliac trunk and its branches. It was done on 46 complexes of the organs from the persons of various age, the cause of death being other than the pathologies of the organs involved. There are described results of our study in order to understand the selection criteria of surgical interventions on different patients with the same pathology. Lucrarea reprezintă un studiu asupra trunchilui celiac şi ramurilor sale, efectuat în baza studierii a 46 complexe de organe de la persoane de vîrste diferite, la care decesul nu a fost provocat de patologii la nivelul organelor implicate. Rezultatele au pus la evidenţă importanţa studierii lor, pentru a înţelege criteriul selecţiei a diferitor intervenţii chirurgicale la diferiţi pacienţi cu aceeaşi patologie

Devitalization of Glioblastoma Cancer Cells by Non-invasive Physical Plasma: Modulation of Proliferative Signalling Cascades

Background/Aim: Glioblastoma (GBM) is the most common and most lethal type of cancer of the central nervous system in adults. Despite aggressive treatment, which is based on surgical resection, if possible, followed by radiation and chemotherapy, a high recurrence rate and therapy resistance is observed. Thus, additional innovative therapies are urgently needed to improve the poor median survival of only 15 months. Treatment of solid tumours with non-invasive physical plasma (NIPP) represents such a novel and innovative anticancer procedure. Materials and Methods: In this study, we investigated the effect of NIPP, an ionized argon gas, on the in vitro growth of human GBM cell lines, LN-18 and U-87 MG. Proliferation was measured by live cell count. Subsequently, proliferative factors were analysed at the level of nucleic acids (polymerase chain reaction) and proteins (western blotting). Results: For both GBM lines, a treatment time-dependent decrease in growth was observed compared to controls. Additionally, NIPP treatment resulted in reduced rates of AKT serine/threonine kinase 1 (AKT1) and extracellular-regulated kinase 1/2 ERK1/2 expression, whereas expression of p21, proliferating cell nuclear antigen, and heat-shock proteins 90α and 90β was not affected. In both cell lines, a strong increase in expression of tumour-suppressive microRNA-1 (miR-1) was detected after exposure to NIPP. Conclusion: Our results demonstrated that NIPP is able to efficiently attenuate growth of GBM cells and suggest AKT1, ERK1/2 and miR-1 to be pivotal factors of NIPP-modulated cellular signalling. Translated into the clinical setting, NIPP may represent a promising option for the treatment of GBM

Non-Invasive Physical Plasma Reduces the Inflammatory Response in Microbially Prestimulated Human Gingival Fibroblasts.

Non-invasive physical plasma (NIPP), an electrically conductive gas, is playing an increasingly important role in medicine due to its antimicrobial and regenerative properties. However, NIPP is not yet well established in dentistry, although it has promising potential, especially for periodontological applications. The aim of the present study was to investigate the effect of NIPP on a commercially available human gingival fibroblast (HGF) cell line and primary HGFs in the presence of periodontitis-associated bacteria. First, primary HGFs from eight patients were characterised by immunofluorescence, and cell numbers were examined by an automatic cell counter over 5 days. Then, HGFs that were preincubated with Fusobacterium nucleatum (F.n.) were treated with NIPP. Afterwards, the IL-6 and IL-8 levels in the cell supernatants were determined by ELISA. In HGFs, F.n. caused a significant increase in IL-6 and IL-8, and this F.n.-induced upregulation of both cytokines was counteracted by NIPP, suggesting a beneficial effect of physical plasma on periodontal cells in a microbial environment. The application of NIPP in periodontal therapy could therefore represent a novel and promising strategy and deserves further investigation

Cold Atmospheric Plasma Treatment of Chondrosarcoma Cells Affects Proliferation and Cell Membrane Permeability

Chondrosarcoma is the second most common malign bone tumor in adults. Surgical resection of the tumor is recommended because of its resistance to clinical treatment such as chemotherapy and radiation therapy. Thus, the prognosis for patients mainly depends on sufficient surgical resection. Due to this, research on alternative therapies is needed. Cold atmospheric plasma (CAP) is an ionized gas that contains various reactive species. Previous studies have shown an anti-oncogenic potential of CAP on different cancer cell types. The current study examined the effects of treatment with CAP on two chondrosarcoma cell lines (CAL-78, SW1353). Through proliferation assay, the cell growth after CAP-treatment was determined. A strong antiproliferative effect for both cell lines was detected. By fluorescein diacetate (FDA) assay and ATP release assay, alterations in the cell membrane and associated translocation of low molecular weight particles through the cytoplasmic membrane were observed. In supernatant, the non-membrane-permeable FDA and endogenously synthesized ATP detected suggest an increased membrane permeability after CAP treatment. Similar results were shown by the dextran-uptake assay. Furthermore, fluorescence microscopic G-/F-actin assay was performed. G-and F-actin were selectively dyed, and the ratio was measured. The presented results indicate CAP-induced changes in cell membrane function and possible alterations in actin-cytoskeleton, which may contribute to the antiproliferative effects of CAP. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

Modulation of the Tumor-Associated Immuno-Environment by Non-Invasive Physical Plasma

Over the past 15 years, investigating the efficacy of non-invasive physical plasma (NIPP) in cancer treatment as a safe oxidative stress inducer has become an active area of research. So far, most studies focused on the NIPP-induced apoptotic death of tumor cells. However, whether NIPP plays a role in the anti-tumor immune responses need to be deciphered in detail. In this review, we summarized the current knowledge of the potential effects of NIPP on immune cells, tumor–immune interactions, and the immunosuppressive tumor microenvironment. In general, relying on their inherent anti-oxidative defense systems, immune cells show a more resistant character than cancer cells in the NIPP-induced apoptosis, which is an important reason why NIPP is considered promising in cancer management. Moreover, NIPP treatment induces immunogenic cell death of cancer cells, leading to maturation of dendritic cells and activation of cytotoxic CD8+ T cells to further eliminate the cancer cells. Some studies also suggest that NIPP treatment may promote anti-tumor immune responses via other mechanisms such as inhibiting tumor angiogenesis and the desmoplasia of tumor stroma. Though more evidence is required, we expect a bright future for applying NIPP in clinical cancer management

Expression of the MIR-1 molecule in patients with uterine leiomyosarcoma

Department of Surgery, University Medicine Greifswald, Ferdinand-Sauerbruch-Strasse, 17475 Greifswald, Germany, The 6th International Medical Congress for Students and Young DoctorsBackground: The uterine leiomyosarcoma represents the most frequent malignant gynecologic mesenchymal tumor that often develops distant metastases. The diagnosis of these tumors is nowadays still a challenge and the direct implication of the small non-coding RNAs (MicroRNAs ) in gene expression, tumor initiation and tumor progression has already been revealed in scientific studies. Because the aberrant microRNA (miRNA) expression patterns show a diagnostic value as tumor markers, we aimed to identify the gene expression level of miRNA-1 (miR-1) and the protein targets in uterine leiomyosarcoma. Methods: Using the specific cell line - SK-UT-1 with similar biological characteristics of the uterine leiomyosarcoma tissue, in comparison to ovarian carcinoma cell lines: OVCAR-3, TOV-21 and SK-OV-3, and cell lines of mouse heart-muscle (HL-1), we were able to perform real time PCRs and RNA-Isolation arrays, transient and stabile transfection programs with lipofectamine reagents. Tissue samples of uterine leiomyosarcoma and healthy uterus were again analyzed by means of transfection and isolation arrays. The electrophoresis using protein targets of the miR-1 (p38 and ERK 1/2 widely expressed protein kinase intracellular signaling molecules and involved in functions including the regulation of meiosis, mitosis, und postmitotic functions) was also integrated. Results: The analysis of the SK-UT-1 cell line have shown significant differences in comparison to the other studied cell lines, respectively a reduced expression of the miR-1 molecules. The same results were observed in the process of transfection and electrophoresis of the human tissues, where the lowest expression of the miR-1 was evidenced in the uterine leiomyosarcomas. The specific protein targets of miR-1 have shown positive Western Blot signals. Conclusions: The miR-1 non coding molecules may improve our understanding of disease development, progression and gene expression of the uterine leiomyosarcoma. Further prospective translational studies in order to evaluate miR-1 as a prognostic factor are needed. Key words: MIR-1, leiomyosarcoma, Western Blot