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Consumers are exposed to mineral oil hydrocarbons (MOH) e.g. through foodstuffs and cosmetics. Upon ingestion, MOH follow the absorption pathway of dietary lipids. Analytical chemistry has revealed the presence of the main fraction, designated as mineral oil saturated hydrocarbons (MOSH), in mesenteric lymph nodes (MLNs), liver, spleen, and adipose tissue. Recent results from animal studies raised concerns about a long-term, possibly irreversible accumulation of some MOSH in humans. To address this issue, we performed a statistical re-analysis of published biopsy and autopsy data regarding the age-dependence of MOSH levels in human tissue. MOSH concentrations in MLNs and adipose tissue showed a 1.2–1.4-fold increase per decade, pointing to very long-term accumulation in both tissues. There was no evidence for age-dependent MOSH concentrations in liver and spleen. There was no sex difference in the MOSH concentrations in MLNs, suggesting a similar oral exposure for men and women. On average, women had a 2.2–2.5-fold higher MOSH concentration in the liver, spleen and adipose tissue compared to men. This finding may point to a sex difference in metabolism, in line with animal data. The use of certain cosmetics was a relevant predictor in addition to age. Women that used cosmetics like lipstick, hand cream, and sun cream had an average 2.1-fold higher MOSH concentration in abdominal subcutaneous fat than non-users.

Clonal hematopoiesis (CH) driven by mutations in the DNA damage response (DDR) pathway is frequent in patients with cancer and is associated with a higher risk of therapy-related myeloid neoplasms (t-MNs). Here, we analyzed 423 serial whole blood and plasma samples from 103 patients with relapsed high-grade ovarian cancer receiving carboplatin, poly(ADP-ribose) polymerase inhibitor (PARPi) and heat shock protein 90 inhibitor (HSP90i) treatment within the phase II EUDARIO trial using error-corrected sequencing of 72 genes. DDR-driven CH was detected in 35% of patients and was associated with longer duration of prior PARPi treatment. TP53- and PPM1D-mutated clones exhibited substantially higher clonal expansion rates than DNMT3A-or TET2-mutated clones during treatment. Expansion of DDR clones correlated with HSP90i exposure across the three study arms and was partially abrogated by the presence of germline mutations related to homologous recombination deficiency. Single-cell DNA sequencing of selected samples revealed clonal exclusivity of DDR mutations, and identified DDR-mutated clones as the origin of t-MN in two investigated cases. Together, these results provide unique insights into the architecture and the preferential selection of DDR-mutated hematopoietic clones under intense DNA-damaging treatment. Specifically, PARPi and HSP90i therapies pose an independent risk for the expansion of DDR-CH in a dose-dependent manner.

TP53is the most commonly mutated gene in cancer and has been shown to form amyloid-like aggregates, similar to key proteins in neurodegenerative diseases. Nonetheless, the clinical implications of p53 aggregation remain unclear. Here, we investigated the presence and clinical relevance of p53 aggregates in serous ovarian cancer (OC). Using the p53-Seprion-ELISA, p53 aggregates were detected in 46 out of 81 patients, with a detection rate of 84.3% in patients with missense mutations. High p53 aggregation was associated with prolonged progression-free survival. We found associations of overall survival with p53 aggregates, but they did not reach statistical significance. Interestingly, p53 aggregation was significantly associated with elevated levels of p53 autoantibodies and increased apoptosis, suggesting that high levels of p53 aggregates may trigger an immune response and/or exert a cytotoxic effect. To conclude, for the first time, we demonstrated that p53 aggregates are an independent prognostic marker in serous OC. P53-targeted therapies based on the amount of these aggregates may improve the patient’s prognosis.

Human papillomavirus antibody reactivities to vulvar precancerous lesions were reported but the role of human papillomavirus in some of the lesions is still unclear. Reports supported the role of high-risk human papillomavirus types, mainly human papillomavirus-16, in the pathogenesis of different vulvar lesions with atypia.

Clonal hematopoiesis (CH), characterized by the expansion of somatically mutated hematopoietic stem cells, is an age-related phenomenon associated with the development of hematologic malignancies, increased cardiovascular risk, and other age-related proinflammatory conditions. In patients with solid tumors, cytotoxic therapies differentially select for clones with mutations in the DNA damage response (DDR) pathway, i.e. TP53and PPM1D, leading to an elevated risk for therapy-related myeloid malignancies (t-MNs). Poly(ADP-ribose) polymerase inhibitors (PARPi) are frequently used to treat ovarian cancer with particularly high efficacy reported in tumors exhibiting homologous recombination (HR) deficiency caused by mutations in BRCA1/2or other HR pathway genes. However, recent data indicate that PARPi treatment is also associated with an elevated risk for t-MNs.

A growing number of diseases are being linked to protein misfolding and amyloid formation. Recently, p53 was also shown to associate into amyloid aggregates, raising the question of whether cancer development is associated with protein aggregation as well. However, a lack of suitable tools has hampered the evaluation of their clinical relevance. Herein, we report an enzyme-linked-immunosorbent-assay (ELISA) system based on a polyionic, high-molecular-weight ligand that specifically captures aggregated oligomers and amyloid proteins. We proved that naturally occurring tetramers of p53 are not bound, but high-molecular-weight aggregates are bound and subsequently detected. For the first time, this assay allows the quantitative detection of p53 aggregates from cell lysates, which was demonstrated using 22 ovarian-cancer cell lines as well as 7 patient-derived tumor tissues. The levels of p53 aggregates within the missense-mutated tissue samples varied more than 12-fold. This simple, robust method allows studying the abundance and clinical relevance of protein aggregates. This could help our understanding of the role of protein misfolding in cancer or even in predicting therapy responses to aggregation-targeting drugs.

All current regimens for treating ovarian cancer center around carboplatin as standard first line. The HSP90 inhibitor ganetespib is currently being assessed in advanced clinical oncology trials. Thus, we tested the combined effects of ganetespib and carboplatin on a panel of 15 human ovarian cancer lines. Strikingly, the two drugs strongly synergized in cytotoxicity in tumor cells lacking wild-type p53. Mechanistically, ganetespib and carboplatin in combination, but not individually, induced persistent DNA damage causing massive global chromosome fragmentation. Live-cell microscopy revealed chromosome fragmentation occurring to a dramatic degree when cells condensed their chromatin in preparation for mitosis, followed by cell death in mitosis or upon aberrant exit from mitosis. HSP90 inhibition caused the rapid decay of key components of the Fanconi anemia pathway required for repair of carboplatin-induced interstrand crosslinks (ICLs), as well as of cell cycle checkpoint mediators. Overexpressing FancA rescued the DNA damage induced by the drug combination, indicating that FancA is indeed a key client of Hsp90 that enables cancer cell survival in the presence of ICLs. Conversely, depletion of nuclease DNA2 prevented chromosomal fragmentation, pointing to an imbalance of defective repair in the face of uncontrolled nuclease activity as mechanistic basis for the observed premitotic DNA fragmentation. Importantly, the drug combination induced robust antitumor activity in xenograft models, again accompanied with depletion of FancA. In sum, our findings indicate that ganetespib strongly potentiates the antitumor efficacy of carboplatin by causing combined inhibition of DNA repair and cell cycle control mechanisms, thus triggering global chromosome disruption, aberrant mitosis and cell death.

L1 cell adhesion molecule (L1CAM) is a transmembrane molecule belonging to the L1 protein family. It has shown to be a key player in axonal guidance in the course of neuronal development. Furthermore, L1CAM is also crucial for the establishment of the enteric and urogenital organs and is aberrantly expressed in cancer originating in these organs. Carcinogenesis and embryogenesis follow a lot of similar molecular pathways, but unfortunately, comprehensive data on L1CAM expression and localization in human developing organs are lacking so far. In the present study we, therefore, examined the spatiotemporal distribution of L1CAM in the early human fetal period (weeks 8–12 of gestation) by means of immunohistochemistry and in situ hybridization (ISH). In the epithelia of the gastrointestinal organs, L1CAM localization cannot be observed in the examined stages most likely due to their advanced polarization and differentiation. Despite these results, our ISH data indicate weak L1CAM expression, but only in few epithelial cells. The genital tracts, however, are distinctly L1CAM positive throughout the entire fetal period. We, therefore, conclude that in embryogenesis L1CAM is crucial for further differentiation of epithelia.

An international joint statement about the use of hyperthermic intraperitoneal chemotherapy (HIPEC) in ovarian cancer was published in 2016, warning about the uncritical use of HIPEC outside controlled studies. This statement has now been updated after the most recent literature was reviewed by the participating study groups and societies. HIPEC became a treatment option in patients with advanced colon cancer after positive results of a randomized trial comparing surgery and HIPEC versus palliative treatment alone. Although this trial did not compare the added value of HIPEC to surgery alone, HIPEC for the treatment of peritoneal metastases was in the subsequent years generalized to many other cancer types associated with peritoneal carcinomatosis including epithelial ovarian cancer (EOC). In the meantime, new evidence from prospective randomized trials specifically for EOC- patients emerged, with however contradicting results and several quality aspects that made the interpretation of their findings critical. Moreover, three additional trials in colorectal cancer failed to confirm the previously presumed survival benefit through the implementation of HIPEC in peritoneally disseminated colorectal cancers. Based on a still unclear and inconsistent landscape, the authors conclude that HIPEC should remain within the remit of clinical trials for EOC- patients. Available evidence is not yet sufficient to justify its broad endorsement into the routine clinical practice.

The p73 gene gives rise to the full-length transactivation competent TAp73 and the N-terminally truncated isoform Np73, which inhibits TAp73 and wild-type p53. The clinical relevance of TAp73 and Np73 protein expression has not yet been evaluated in ovarian cancer. TAp73 and Np73 expression was examined using immunohistochemistry and reverse transcription–polymerase chain reaction in 83 and 64 ovarian cancer specimens, respectively. A yeast-based assay and subsequent sequencing were performed to analyze the p53mutational status. TAp73 and Np73 protein expression was found in 73 of 83 (88) and 48 of 83 (57.8) ovarian cancer samples, respectively. The majority of cases showed immunostaining in both the nucleus and cytoplasm of tumor cells. TAp73 and Np73 protein expression correlated with messenger RNA quantification in 25 of 64 (39.1) and 37 of 64 (57.8) cancer specimens, respectively. TAp73 and Np73 protein expression was not associated with the p53mutational status, clinicopathologic parameters, and prognosis of the examined ovarian cancer cases. Although TAp73 and Np73 protein expression did not possess prognostic significance for ovarian cancer in this study, a potential clinical role of p73 isoforms cannot be definitively excluded due to limitations of immunohistochemistry.

There is growing evidence that deregulation of E2F transcription factors is causatively involved in the patho-physiology of various tumors. However, no data on the role of E2F family members in tumor biology of ovarian cancer are available. We here describe an expression study of all known E2F transcription factors and their co-activators DP-1 and DP-2 in various human ovarian cancer cell lines and the breast cancer cell line T47D and their involvement in pathways affected by interferon-g and EGF. A significant over-expression of the proliferation-promoting E2F1 and especially E2F2 points to a pivotal role in modulating the uncontrolled proliferation in ovarian cancer cells. Of special note is the fact that interferon-g treatment did not only caused a reduction of the proliferation-promoting transcription factors E2F1 and E2F2, but also increased the inhibiting transcription factors E2F4 and E2F5, thus underlining the importance of an E2F cross-talk in the anti-proliferative function of interferon-g. Moreover, an increase in DP-1 is probably involved in the proliferation-enhancing effect of EGF. Our study provides a new insight in the crucial role of E2F cross-talk, especially the role of the inhibiting transcription factors E2F4 and E2F5, in the tumor biology of cancer and its possible usefulness as targets in anti-cancer therapy.

We analyzed the p53mutational status, mRNA and protein expression in 24 human breast carcinoma cell lines. Following measurement of their DNA content with flow cytometry, we ascertained the copy numbers of the centromere of chromosome 17 (cen17) and p53with fluorescence in situhybridization (FISH). A functional yeast assay (FASAY) was used to screen for inactivating mutations. Positive results were subsequently verified by DNA sequencing. Finally, we assessed the mRNA expression with a competitive reverse transcription-polymerase chain reaction (RT-PCR) assay and the protein expression with immunocytochemical staining, western blot, and quantitative flow cytometry. The DNA content of the cell lines ranged from 0.85 to 2.58. Nine cell lines had concordant copy numbers (between two and four) of p53and cen17, whereas 12 had more, and three less cen17 than p53copies. The FASAY was successful in all but one cell line and revealed the presence of mutated alleles in 16 of them, 13 cell lines expressed only the mutated, and three both the mutated and the wild-type alleles. The mutations were comprised of 11 missense, two nonsense, and three frameshift mutations. Immunocytochemical staining, western blot and quantitative flow cytometry yielded comparable p53 protein expression results. However, both the mRNA and the protein expression levels varied considerably in the different cell lines and no consistent pattern with regard to the respective p53mutational status became evident. The results obtained in these breast carcinoma cell lines indicate that no clear-cut linear relationship exists between the p53mutational status and the extent of its respective mRNA and protein expression. Therefore, direct DNA analyses and functional assays remain the only methods for the reliable detection of p53mutations.

The aim of the present study was to evaluate the clinical usefulness of the cytokeratin tumor marker tissue polypeptide antigen (TPA) in patients with vulvar cancer. This retrospective study comprises 41 patients with vulvar cancer FIGO stages I–III, 17 patients with vulvar intraepithelial neoplasia (VIN) III, and 40 healthy female controls. Serum concentrations of TPA were measured using a microparticle enzyme immunoassay. Results were correlated to clinical data. Median serum concentrations of TPA in healthy female controls, patients with VIN III, and patients with vulvar cancer were 42 U/l (range 12–192), 53 U/l (range 17–127.9) and 57 U/l (range 4.2–423), respectively (Mann-Whitney U test, p = 0.8). Serum concentrations of TPA were not associated with stage of disease, histological grade, and age at the time of diagnosis. In vulvar cancer patients, elevated serum concentrations of TPA prior to therapy were not associated with a shortened disease-free or overall survival (log-rank test: p = 0.5 and p = 0.9, respectively). In a multivariate Cox regression model comprising tumor stage and TPA, tumor stage, but not TPA revealed a statistically significant influence on disease-free (Cox proportional hazard regression model, p = 0.05 and p = 0.6, respectively) and overall (Cox proportional hazard regression model, p = 0.04 and p = 0.8, respectively) survival of patients with vulvar cancer. We conclude that cytokeratin expression, as reflected by serum concentrations of TPA, does not play a role in the natural history of vulvar cancer. The evaluation of serum concentrations of TPA prior to therapy is not recommended.

The invasive potential of tumor cells is usually tested either by in vitro invasion assays which evaluate cell spreading ability in basement membrane‐like matrices or by in vivo invasion assays in nude mice. Both methods are laborious and time‐consuming. Tumor invasiveness is accompanied by the changes in expression of various genes. The invasive behavior of cells is therefore represented by certain gene expression patterns. The purpose of this study was to investigate whether expression patterns of several genes are characteristic for the invasiveness of cultured cells. We examined the mRNA levels of estrogen receptor (ER), progesterone receptor (PR), estrogen inducible pS2 and plasminogen activator inhibitor‐1 (PAI‐1) in 23 cell lines derived from benign and malignant breast tissues using a competitive reverse transcription‐polymerase chain reaction (cRT‐PCR) system. We also evaluated the invasiveness of these cell lines by their ability to penetrate into a collagen‐fibroblast matrix. We demonstrate that the gene expression pattern of breast cell lines is clearly associated with its in vitro invasiveness. In general, cells with ER, PR, pS2 but no PAI‐1 expression showed a non‐invasive phenotype, while cells expressing PAI‐1 mRNA but not ER mRNA are invasive. Our study indicates that the invasiveness of breast cancer cell lines is characterized by PAI‐1 gene expression and the lack of ER mRNA. This suggests that PAI‐1 may participate in the invasive process.

Since publication of this article, the authors reported that the online version is missing the links to most of the Supplementary data, specifically, Supplementary Figures S1–S9; Supplementary Table S1; all legends to Supplementary Material.