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1.
Pathol Oncol Res ; 22(4): 741-6, 2016 Oct.
Article in English | MEDLINE | ID: mdl-27075652

ABSTRACT

The hypoxia that arises due to the rapid proliferation of tumor cells is a fundamental driving force for the canonical pathway of neovascularization. In the current study we report a very strong correlation between mRNA expression levels of HIF-2α (but not HIF-1α), VEGFR-1, VEGFR-2 and MMP2 in ex vivo samples from laryngeal carcinoma. Sixty-three samples from patients with histopathologically verified carcinoma of the larynx were examined in this study. Total RNA was isolated from both normal and tumor fresh frozen tissues of each patient and real-time quantitative PCR reactions were performed. The mRNA expression levels of HIF-1α, HIF-2α, VEGFR1, VEGFR2 and MMP2 were acquired. We found strong positive correlations between mRNA expression levels of HIF-2α and VEGFR-1, r s (98) = .671, p < .0005; HIF-2α and VEGFR-2, r s (98) = .742, p < .0005; HIF-2α and MMP2, r s (98) = .566, p < .0005; VEGFR-1 and VEGFR-2, r s (98) = .791, p < .0005; VEGFR-1 and MMP2, r s (98) = .709, p < .0005; VEGFR-2 and MMP2, r s (98) = .793, p < .0005. Our results provide evidence for the regulatory connection between HIF-2α and VEGFR-1, VEGFR-2 and MMP2 in the light of ETS1/ HIF-2α regulatory axis on a non-in-vitro level in carcinoma tissue, uncover some of the differences between the homologues HIF-1α and HIF-2α and round up and support the results from different experimental models in this field.


Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Carcinoma/genetics , Laryngeal Neoplasms/genetics , Matrix Metalloproteinase 2/genetics , RNA, Messenger/genetics , Vascular Endothelial Growth Factor Receptor-1/genetics , Vascular Endothelial Growth Factor Receptor-2/genetics , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged
2.
Ann Oncol ; 27(3): 532-9, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26681675

ABSTRACT

BACKGROUND: Next-generation sequencing (NGS) of tumour samples is a critical component of personalised cancer treatment, but it requires high-quality DNA samples. Routine neutral-buffered formalin (NBF) fixation has detrimental effects on nucleic acids, causing low yields, as well as fragmentation and DNA base changes, leading to significant artefacts. PATIENTS AND METHODS: We have carried out a detailed comparison of DNA quality from matched samples isolated from high-grade serous ovarian cancers from 16 patients fixed in methanol and NBF. These experiments use tumour fragments and mock biopsies to simulate routine practice, ensuring that results are applicable to standard clinical biopsies. RESULTS: Using matched snap-frozen tissue as gold standard comparator, we show that methanol-based fixation has significant benefits over NBF, with greater DNA yield, longer fragment size and more accurate copy-number calling using shallow whole-genome sequencing (WGS). These data also provide a new approach to understand and quantify artefactual effects of fixation using non-negative matrix factorisation to analyse mutational spectra from targeted and WGS data. CONCLUSION: We strongly recommend the adoption of methanol fixation for sample collection strategies in new clinical trials. This approach is immediately available, is logistically simple and can offer cheaper and more reliable mutation calling than traditional NBF fixation.


Subject(s)
DNA/drug effects , Formaldehyde/chemistry , Methanol/chemistry , Neoplasms/diagnosis , Tissue Fixation/methods , Base Sequence , DNA/analysis , High-Throughput Nucleotide Sequencing , Humans , Middle Aged , Paraffin Embedding , Sequence Analysis, DNA
3.
Neoplasma ; 62(1): 27-33, 2015.
Article in English | MEDLINE | ID: mdl-25563364

ABSTRACT

UNLABELLED: Saponins are natural substances produced by a large number of plants, one of which is Tribulus terrestris L. (TT). They have been reported to possess an antitumor activity exerted by regulating various signaling pathways in the cell. Although the mechanisms of action of saponin extracts from various plants have been widely studied, limited data are available about TT. The present study aimed to analyze the impact of saponin extract from TT on cell processes in breast carcinoma cell lines. The variations in expression of a group of 32 selected genes were examined by real-time PCR after saponin treatment of MCF7 and MCF10A cell lines. Only three genes - CXCR4, CCR7 and BCL2, showed changes in their mRNA levels after the application of the herb extract. While CXCR4 expression was reduced in both cell lines, CCR7 and BCL2 levels decreased only in tumorigenic MCF7 cells, implying cell-specificity of the saponin action. Our results suggested that TT extract containing saponins was likely to affect the processes of apoptosis and metastasizing of cancer cells. Further in vivo studies will show its applicability as an anticancer therapeutic agent. KEYWORDS: saponins, Tribulus terrestris, breast cancer, CXCR4, CCR7, BCL2.

4.
Acta Otorhinolaryngol Ital ; 35(5): 321-4, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26824913

ABSTRACT

Angiogenesis is one of the six originally constituted hallmarks of cancer that has been extensively studied in the last two decades. The aim of our study is to assess the microvessel and macrophageal density in laryngeal carcinoma and its clinicopathological correlations. We immunohistochemically assessed microvessel density (CD34) and macrophage count (CD68) using microarray techniques and then looked for clinicopathological correlations. The mean micro-vessel density in the study group was 14.27 ± 12.92 vessels in a ×200 field with a mean macrophageal infiltration density of 5.19 ± 4.32. Median microvessel density was significantly higher in patients with metastasis than in patients without metastasis. Additionally, linear regression established that macrophageal infiltration density could predict microvessel density in laryngeal carcinoma. We found no association between either factor and recurrence rate or other clinical characteristics. Our study adds additional data to a problem that has been widely studied during the last two decades, even if controversies in this area still remain.


Subject(s)
Laryngeal Neoplasms/blood supply , Microvessels , Neovascularization, Pathologic , Humans , Immunohistochemistry , Neoplasm Recurrence, Local , Prognosis
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