Aneuploid serves as a prognostic marker and favors immunosuppressive microenvironment in ovarian cancer.

Ovarian cancer is the most lethal gynecologic neoplasm, and most patients experience recurrence and chemoresistance. Even the promising immunotherapy showed limited efficacy in ovarian cancer, probably due to the immunosuppressive microenvironment. However, the behind mechanisms of the immune exclusion or cold phenotype in ovarian cancer still remain to be explored. As a cancer dominated by copy number variations instead of mutations, ovarian cancer contains a high fraction of aneuploid, which might correlate with immune inhibition. Nevertheless, whether or how aneuploid affects ovarian cancer is still unclear. For exploring the role of aneuploid cancer cells and the potential ploidy-immune relationship, herein, the ploidy information was first comprehensively analyzed combining the karyotype data and copy number variation data obtained from Mitelman and cBioPortal databases, respectively. Ovarian cancer showed strong ploidy heterogeneity, with high fraction of aneuploid and recurrent arm-level and whole chromosome changes. Furthermore, clinical parameters were compared between the highly-aneuploid and the near-diploid ovarian cancers. Aneuploid indicated high grade, poor overall survival and poor disease-free survival in ovarian cancer. To understand the biofunction affected by aneuploid, the differentially expressed genes between the highly-aneuploid and the near-diploid groups were analyzed. Transcription data suggested that aneuploid cancer correlated with deregulated MHC expression, abnormal antigen presentation, and less infiltration of macrophages and activated T cells and higher level of T cell exclusion. Furthermore, the ploidy-MHC association was verified using the Human Protein Atlas database. All these data supported that aneuploid might be promising for cancer management and immune surveillance in ovarian cancer.

Ploidy Status of Ovarian Cancer Cell Lines and Their Association with Gene Expression Profiles.

As a cancer type potentially dominated by copy number variations, ovarian cancer shows hyperploid karyotypes and large-scale chromosome alterations, which might be promising biomarkers correlated with tumor metastasis and chemoresistance. Experimental studies have provided more information about the roles of aneuploids and polyploids in ovarian cancer. However, ploidy evaluation of ovarian cancer cell lines is still limited, even in some ploidy-related research. Herein, the ploidy landscape of 51 ovarian cancer cell lines from the Cancer Cell Line Encyclopedia (CCLE) were analyzed, and the ploidy statuses of 13 human ovarian cancer cell lines and 2 murine cell lines were evaluated using G-banding and flow cytometry. Most human ovarian cancer cell lines were aneuploid, with modal numbers of 52-86 and numerical complexity ranging from 5 to 12. A2780, COV434 and TOV21G were screened as diploid cell lines, with a modal number of 46, a low aneuploid score and a near-diploid ploidy value. Two murine cell lines, both OV2944-HM1 and ID-8, were near-tetraploid. Integrated information on karyotypes, aneuploid score and ploidy value supplied references for a nondiploid model construction and a parallel analysis of diploid versus aneuploid. Moreover, the gene expression profiles were compared between diploid and aneuploid cell lines. The functions of differentially expressed genes were mainly enriched in terms of protein function regulation, TGF-ß signaling and cell adhesion molecules. Genes downregulated in the aneuploid group were mainly related to metabolism and protein function regulation, and genes upregulated in the aneuploid group were mainly involved in immune regulation. Differentially expressed genes were randomly distributed on all chromosomes, while chromosome 1 alteration might contribute to immune-related alterations in aneuploid cell lines. Chromosome 19 alteration might be potentially significant for aneuploid ovarian cancer cell lines and patients, which needs further verification in ploidy research.

Detection of Genetic Alterations in Oral Squamous Cell Carcinoma Using Multiplex LigationDependent Probe Amplification (MLPA)

@#Deletions and amplifications of genes often occur during multistep progression from oral precancer, seen as oral epithelial dysplasia (OED) to cancerous stage. These genetic alterations could be used as markers to aid in detection of oral squamous cell carcinomas (OSCC). This study explored the use of multiplex ligation-dependent probe amplification (MLPA) technique in detecting OSCC and OED specific genetic alterations. MLPA was used to detect gains and losses of 106 genes in DNA extracted from frozen tissue samples of 10 OSCC and 10 noncancer patients. Two biopsies of OED were analyzed to explore the alterations in oral potentially malignant disorders. There were significant differences (p<0.001) in the number of alterations in OSCC and dysplasia compared to non-cancer samples respectively. The most frequently altered genes in OSCC were PTP4A3, RECQL4, ATM, and KLK3 (60%). Five genes (MYC, SLA, TNFRSF1A, MESDC1, MIF) were altered in 50% of OSCC samples. These nine genes were specific to OSCC samples (p<0.05). Some genes, including MYB, MET, CASP2, SLA and PTEN occurred in 50% of OED samples. MLPA was able to detect genetic alterations, that are present only in the OSCC samples and showed potential to be used as an adjunctive tool in early diagnosis of OSCC.

The Role of Genetic Analysis in Correct Diagnosis of Eosinophilic Variant of Chromophobe Renal Cell Carcinoma.

BACKGROUND/AIM: It has been suggested that eosinophilic variant of chromophobe renal cell carcinoma (chRCC) with low chromosome number or lack of genomic alteration has an excellent prognosis in comparison to classic chRCC. The aim of our study was to analyse the phenotypical variations of 77 chRCCs, including 7 eosinophilic ones, each diagnosed unequivocally by genetic means. MATERIALS AND METHODS: DNA isolated from chRCCs was subjected to array comparative genomic hybridisation (CGH) for establishing the chromosome alteration. Original histological slides were evaluated for cellular phenotype and growth pattern and compared to the genetic alterations. RESULTS: Loss of the entire chromosome 1, 2, 6, 10, 13, 17 and 21 occurred in 95%, 94%, 86% 90% 82% 90% and 66% of the cases, respectively. The number of chromosome alterations in eosinophilic forms of chRCC corresponded to those found in classic chRCC with pale-reticular cytoplasm or mixed cellular characteristics. Three of seven eosinophilic variants with loss of 4, 10 and 11 chromosomes showed metastasis at the time of diagnosis whereas only 3 metastatic tumors were noticed among the 70 classic chRCC. We did not find discriminating difference in number of chromosome alteration between classic and eosinophilic forms of chRCC. CONCLUSION: Eosinophilic chRCC has a more aggressive biology than the classic form. To avoid diagnostic pitfall of eosinophilic renal cell tumors with uncertain diagnosis, a genetic analysis should be carried out.

Citotoxicidade das águas do Rio do Peixe (São Paulo-Brasil), em células meristemáticas de raiz de Allium cepa L / Cytotoxicity of waters of the River Peixe (São Paulo-Brazil), in meristematic root cells of Allium cepa L

Nas últimas décadas, a qualidade da água do Rio do Peixe, localizado no interior do Estado de São Paulo, está diminuindo, principalmente porque o rio vem sofrendo com o desmatamento feito às suas margens, o que provocou o assoreamento e permitiu o despejo de todos os tipos de resíduos, em especial das indústrias que ficam ao seu redor. Assim, o objetivo deste trabalho foi avaliar o potencial citotóxico das águas do Rio do Peixe, coletadas em quatro propriedades rurais localizadas em Garça­SP (nascente), Tupã-SP, Flórida Paulista-SP e Ouro Verde-SP (jusante), usando como sistema teste vegetal as células meristemáticas de raiz de Allium cepa L. Os resultados obtidos mostraram que todas as amostras das águas coletadas não apresentaram efeito citotóxico após 24 horas de tratamento. Entretanto, as amostras obtidas em Tupã, Flórida Paulista e Ouro Verde (jusante) foram mutagênicas, pois causaram o aparecimento de alterações nas células, principalmente, dos tipos metáfases-colchicínicas, anáfases desorganizadas, multipolares e com ponte cromatídica, além de células micronucleadas. Portanto, os resultados obtidos alertam para o perigo eminente que os efluentes lançados indiscriminadamente no ambiente, de rios e lagos, podem representar não só aos organismos que ali habitam, mas a todos os outros que dele dependem, seja para lazer, limpeza, higiene ou alimentação.

(Cytotoxicity of waters of the River Peixe (São Paulo-Brazil), in meristematic root cells of Allium cepa L.) In recent decades, the water quality of the River Peixe, located within the State of São Paulo, has been decreasing, mainly because the river has suffered from deforestation made to its banks, causing silting and allowed the dumping of all types of waste, especially in industries that are around you. The objective of this study was to evaluate the cytotoxic potential of the waters of the River Peixe, collected at four farms located in Garça-SP (source), Tupã-SP, Flórida Paulista-SP and Ouro Verde-SP (downstream), using as a system test plant root meristematic cells of Allium cepa L. The results showed that all water samples collected was no cytotoxic effect after 24 hours of treatment. However, samples taken in Tupã, Flórida Paulista and Ouro Verde (downstream) were mutagenic, because it caused the appearance of changes in cells, mainly types of colchicine-metaphase, disorganized anaphase, multipolar and with chromatid bridge, addition of micronucleated cells. Therefore, the results warn of imminent danger that the effluents discharged indiscriminately into the environment may represent not only the organisms that live there, but for all others who depend on it, whether for leisure, cleaning, hygiene or food.