NuMA Overexpression in Epithelial Ovarian Cancer

Highly aneuploid tumours are common in epithelial ovarian cancers (EOC). We investigated whether NuMA expression was associated with this phenomenon

Expression analysis of the MCPH1/BRIT1 and BRCA1 tumor suppressor genes and telomerase splice variants in epithelial ovarian cancer.

Aims The aim of this study was to explore the correlation of hTERT splice variant expression with MCPH1/BRIT1 and BRCA1 expression in epithelial ovarian cancer (EOC) samples. Background Telomerase activation can contribute to the progression of tumors and the development of cancer. However, the regulation of telomerase activity remains unclear. MCPH1 (also known as BRIT1, BRCT-repeat inhibitor of hTERT expression) and BRCA1 are tumor suppressor genes that have been linked to telomerase expression. Methods qPCR was used to investigate telomerase splice variants, MCPH1/BRIT1 and BRCA1 expression in EOC tissue and primary cultures. Results The wild type α+/β+ hTERT variant was the most common splice variant in the EOC samples, followed by α+/β− hTERT, a dominant negative regulator of telomerase activity. EOC samples expressing high total hTERT demonstrated significantly lower MCPH1/BRIT1 expression in both tissue (p = 0.05) and primary cultures (p = 0.03). We identified a negative correlation between MCPH1/BRIT1 and α+/β+ hTERT (p = 0.04), and a strong positive association between MCPH1/BRIT1 and both α−/β+ hTERT and α−/β− hTERT (both p = 0.02). A positive association was observed between BRCA1 and α−/β+ hTERT and α−/β− hTERT expression (p = 0.003 and p = 0.04, respectively). Conclusions These findings support a regulatory effect of MCPH1/BRIT1 and BRCA1 on telomerase activity, particularly the negative association between MCPH1/BRIT1 and the functional form of hTERT (α+/β+)

Cross-Country Comparison of Bans on Internet Tobacco Advertising, and Search Interest in Vaping Products

Background: The rise in vaping use across high-income countries is driven partly by marketing, advertising, and flavors promoted by e-cigarette producer firms. The current study aim is to examine the potential variation in the relative research volume on vaping products between countries with highest level of tobacco banning internet advertising and those with no ban on tobacco internet advertising. Method: We used weekly relative search volume (RSV) data produced by Google Trends (GT). A total of eight countries included in the study, Saudi Arabia, United Arab Emirates, United States, Ireland, New Zealand, Canada, United Kingdom, and Australia. The countries were regrouped into ban in internet advertising countries, and the no bans in internet advertising countries. Results: The trend test indicates a statistically significant upward trend in GT vaping search volume across all the included countries. However, the percentage increase of search volume for the ads no ban group was higher than ban group. There is a statistically significant difference in median between the two group, ( P < 0.0007), (median, 6.25 [IQR, 0.103] for the ban in internet advertising countries vs (median 7.5 Interquartile Ranges (IQR), [0.542] for the no bans on internet advertising countries. Conclusion: Countries with stringent bans on online tobacco advertising have shown lower levels of vaping-related search interest, indicating the potential effectiveness of such regulations. Our research underscores the importance of emphasizing comprehensive bans on tobacco advertising and sales online could help mitigate the upward trend in vaping interest

Analysis of NuMA expression in EOC.

<p>A. Statistical analysis of the large ovarian cancer TMA showing NuMA expression increases with disease stage in the mucinous subtype. B. NuMA expression is associated with lymph node involvement (T3cN1MO) (all subtypes). C. NuMA expression is associated with patient age (all subtypes). D. High levels of NuMA decrease with tumour grade in the serous subtype. E. High NuMA levels are associated with stage 4 disease in serous EOCs. F. High NuMA levels increase with tumour grade in mucinous EOCs. G. High NuMA levels decrease with disease stage in endometrioid EOC. H. High NuMA levels decrease with tumour invasion status (T1 to T3) (all subtypes).</p

Recommendations for sample selection, collection and preparation for NMR-based metabolomics studies of blood

Background Metabolic profiling of blood metabolites, particularly in plasma and serum, is vital for studying human diseases, human conditions, drug interventions and toxicology. The clinical significance of blood arises from its close ties to all human cells and facile accessibility. However, patient-specific variables such as age, sex, diet, lifestyle and health status, along with pre-analytical conditions (sample handling, storage, etc.), can significantly affect metabolomic measurements in whole blood, plasma, or serum studies. These factors, referred to as confounders, must be mitigated to reveal genuine metabolic changes due to illness or intervention onset. Review objective This review aims to aid metabolomics researchers in collecting reliable, standardized datasets for NMR-based blood (whole/serum/plasma) metabolomics. The goal is to reduce the impact of confounding factors and enhance inter-laboratory comparability, enabling more meaningful outcomes in metabolomics studies. Key concepts This review outlines the main factors affecting blood metabolite levels and offers practical suggestions for what to measure and expect, how to mitigate confounding factors, how to properly prepare, handle and store blood, plasma and serum biosamples and how to report data in targeted NMR-based metabolomics studies of blood, plasma and serum.Full Tex

NuMA expression assessed by immunofluorescence reveals that NuMA localises to the nucleus during interphase and to the spindle poles during mitosis in cells of ovarian cancer cell lines and primary ascites cultures and fluorescence intensities vary among the primary cultures.

<p>A. Examples of interphase nuclei with low (a-d) and high (e-h) NuMA levels, and mitotic spindles with low (i-l) and high (m-p) NuMA levels at spindle poles. Green – NuMA, red – α-tubulin, blue - DAPI. Scale bar  = 5 µm. B. Histogram comparing NuMA nuclear fluorescence staining intensities in four ovarian cell lines, one cell line established from a primary culture (GYNA0089) and 23 primary cultures derived from ascitic fluid. C. Histogram comparing NuMA staining intensities at spindle poles in the subset of cell lines and cultures where a sufficient number of mitotic cells were for analysis.</p

NuMA expression in ovarian cancer cell lines, primary cultures and tissue.

<p>A. Affymetrix data showing NuMA over-expression in 6 ovarian cell lines (JAMA-2, IOVE (additional immortalized ovarian epithelial cell line), SKOV-3, TR175, OVCA-433 (in addition) and 1847 (all grey bars, designated CL), 38 samples of primary cells cultured from ascitic fluid (black bars, designated – A, including some samples with consecutive collections or passages) and 4 primary cultures established from tumour tissue (designated –T1 to – T4). A1, A2 and A4 are tumour tissue (T1, T2, T4) associated ascites samples. Intensity values obtained for the samples were normalised to an internal control (normal epithelial ovarian tissue (N)) and gene expression levels are shown as the ratio of intensity levels/control intensity level as a log10 value. B. NuMA levels in cell lysates. A representative example of a lysates analysed for NuMA expression by slot blot Black asterix indicates ovarian cell lines (1847, TR175, JAMA-2), brown asterix the neuroblastoma cell line SH-SY5Y, pink asterix the colon adenocarcinoma cell line SW480, blue asterix the cervical cancer cell line HeLa, and yellow asterix the breast cancer cell line MCF7. All other samples are lysates from ovarian primary cultures. C. Histogram comparing NuMA levels in ovarian cell lines, a primary cell line derived from a benign gynaecological disorder (1153-A1) and primary cultures after normalisation against α-tubulin. D. Section of normal (N) ovarian epithelial and stromal tissue and matched ovarian cancer (Ca) at x10 and x40 magnification after NuMA immunostaining. Arrows indicate epithelial cells. Low to medium levels of nuclear NuMA staining can be seen in the normal epithelial cells, with higher levels of nuclear NuMA staining in the ovarian cancer epithelial cells. E. Analysis of a small ovarian TMA shows that NuMA staining intensity increases with grade in ovarian tumour tissue.</p