Tissue-engineered cells producing complex recombinant proteins inhibit ovarian cancer in vivo

Techniques of tissue engineering and cell and molecular biology were used to create a biodegradable scaffold for transfected cells to produce complex proteins. Mullerian Inhibiting Substance (MIS) causes regression of Mullerian ducts in the mammalian embryo. MIS also causes regression in vitro of ovarian tumor cell lines and primary cells from ovarian carcinomas, which derive from Mullerian structures. In a strategy to circumvent the complicated purification protocols for MIS, Chinese hamster ovary cells transfected with the human MIS gene were seeded onto biodegradable polymers of polyglycolic acid fibers and secretion of MIS confirmed. The polymer-cell graft was implanted into the right ovarian pedicle of severe combined immunodeficient mice. Serum MIS in the mice rose to supraphysiologic levels over time. One week after implantation of the polymer-cell graft, IGROV-1 human tumors were implanted under the renal capsule of the left kidney. Growth of the IGROV-1 tumors was significantly inhibited in the animals with a polymer-cell graft of MIS-producing cells, compared with controls. This novel MIS delivery system could have broader applications for other inhibitory agents not amenable to efficient purification and provides in vivo evidence for a role of MIS in the treatment of ovarian cancer.

Estrogens increase the expression of fibulin-1, an extracellular matrix protein secreted by human ovarian cancer cells.

Ovarian cancers have a high ability to invade the peritoneal cavity and some are stimulated by estrogens. In an attempt to understand the mode of action of estrogens on these cancer cells and to develop new markers, we have characterized estrogen-regulated proteins. This study was aimed at identifying a protein secreted by ovarian cancer cells whose level was increased by estradiol [Galtier-Dereure, F., Capony, F., Maudelonde, T. & Rochefort, H. (1992) J. Clin. Endocrinol. Metab. 75, 1497-1502]. By using microprotein sequencing, the 110-kDa protein was identified as fibulin-1, a protein of the extracellular matrix that binds to fibronectin, laminin, and nidogen. The amount of immunoprecipitated fibulin-1 secreted into the medium and present in the cell extract was increased up to 10-fold by estradiol in three estrogen-responsive ovarian cancer cell lines. By immunohistochemistry fibulin-1 was located in the stroma of several ovarian cancers and cysts. The findings highlight a potential role for fibulin-1 in the spread of ovarian cancer in the peritoneal cavity and/or in distal metastases.

Investigation on Heart Rate Variability (HRV) and psycho-social-affective adjustment in Italian women with ovarian cancer

INTRODUZIONE: L’integrazione mente-corpo applicata ad un ambito patologico predominante in questi tempi, come il cancro, è il nucleo di questa tesi. Il background teorico entro cui è inserita, è quello della Psiconeuroendocrinoimmunologia (Bottaccioli, 1995) e Psico-Oncologia. Sono state identificate, nella letteratura scientifica, le connessioni tra stati psicologici (mente) e condizioni fisiologiche (corpo). Le variabili emerse come potenzialmente protettive in pazienti che si trovano ad affrontare il cancro sono: il supporto sociale, l’immagine corporea, il coping e la Qualità della Vita, insieme all’indice fisiologico Heart Rate Variability (HRV; Shaffer & Venner, 2013). Il potenziale meccanismo della connessione tra queste variabili potrebbe essere spiegato dall’azione del Nervo Vago, come esposto nella Teoria Polivagale di Stephen Porges (2007; 2009). OBIETTIVI: Gli obiettivi principali di questo studio sono: 1. Valutare l’adattamento psicologico alla patologia in termini di supporto sociale percepito, immagine corporea, coping prevalente e qualità della vita in donne con cancro ovarico; 2. Valutare i valori di base HRV in queste donne; 3. Osservare se livelli più elevati di HRV sono associati ad un migliore adattamento psicologico alla patologia; 4. Osservare se una peggiore percezione dell’immagine corporea e l’utilizzo di strategie di coping disadattive sono associate ad una Qualità della Vita più scarsa. METODO: 38 donne affette da cancro ovarico, al momento della valutazione libere da patologia, sono state reclutate presso la clinica oncologica del reparto di Ginecologia dell’Azienda Ospedaliero-Universitaria di Parma, Italia. Ad ogni partecipante è stato chiesto di compilare una batteria di test composta da: MSPSS, per la valutazione del supporto sociale percepito; DAS-59, per la valutazione dell’immagine corporea; MAC, per la valutazione delle strategie di coping prevalenti utilizzate verso il cancro; EORTC-QLQ30, per la valutazione della Qualità della Vita. Per ogni partecipante è stato registrato HRV di base utilizzando lo strumento emWave (HeartMath). RISULTATI PRINCIPALI: Rispondendo agli obiettivi 1 e 2, in queste donne si è rilevato una alto tasso di supporto sociale percepito, in particolare ricevuto dalla persona di riferimento. L’area rivelatasi più critica nel supporto sociale è quella degli amici. Per quanto riguarda l’immagine corporea, la porzione di campione dai 30 ai 61 anni, ha delle preoccupazioni globali legate all’immagine corporea paragonabili ai dati provenienti dalla popolazione generale con preoccupazioni riguardo l’aspetto corporeo. Invece, nella porzione di campione dai 61 anni in su, il pattern di disagio verso l’aspetto fisico sembra decisamente peggiorare. Inoltre, in questo campione, si è rilevato un disagio globale verso l’immagine corporea significativamente più alto rispetto ai valori normativi presenti in letteratura riferiti a donne con cancro al seno con o senza mastectomia (rispettivamente t(94)= -4.78; p<0.000001; t(110)= -6.81;p<0.000001). La strategia di coping più utilizzata da queste donne è lo spirito combattivo, seguito dal fatalismo. Questo campione riporta, inoltre, una Qualità della Vita complessivamente soddisfacente, con un buon livello di funzionamento sociale. L’area di funzionalità più critica risulta essere il funzionamento emotivo. Considerando i sintomi prevalenti, i più riferiti sono affaticamento, disturbi del sonno e dolore. Per definire, invece, il pattern HRV, sono stati confrontati i dati del campione con quelli presenti in letteratura, riguardanti donne con cancro ovarico. Il campione valutato in questo studio, ha un HRV SDNN (Me=28.2ms) significativamente più alto dell’altro gruppo. Tuttavia, confrontando il valore medio di questo campione con i dati normativi sulla popolazione sana (Me=50ms), i nostri valori risultano drasticamente più bassi. In ultimo, donne che hanno ricevuto diagnosi di cancro ovarico in età fertile, sembrano avere maggiore HRV, migliore funzionamento emotivo e minore sintomatologia rispetto alle donne che hanno ricevuto diagnosi non in età fertile. Focalizzando l’attenzione sulla ricerca di relazioni significative tra le variabili in esame (obiettivo 3 e 4) sono state trovate numerose correlazioni significative tra: l’età e HRV, supporto percepito , Qualità della Vita; Qualità della Vita e immagine corporea, supporto sociale, strategie di coping; strategie di coping e immagine corporea, supporto sociale; immagine corporea e supporto sociale; HRV e supporto sociale, Qualità della Vita. Per verificare la possibile connessione causale tra le variabili considerate, sono state applicate regressioni lineari semplici e multiple per verificare la bontà del modello teorico. Si è rilevato che HRV è significativamente positivamente influenzata dal supporto percepito dalla figura di riferimento, dal funzionamento di ruolo, dall’immagine corporea totale. Invece risulta negativamente influenzata dal supporto percepito dagli amici e dall’uso di strategie di coping evitanti . La qualità della vita è positivamente influenzata da: l’immagine corporea globale e l’utilizzo del fatalismo come strategia di coping prevalente. Il funzionamento emotivo è influenzato dal supporto percepito dalla figura di riferimento e dal fatalismo. DISCUSSIONI E CONCLUSIONI: Il campione Italiano valutato, sembra essere a metà strada nell’adattamento dello stato psicologico e dell’equilibrio neurovegetativo al cancro. Sicuramente queste donne vivono una vita accettabile, in quanto sopravvissute al cancro, ma sembra anche che portino con sé preoccupazioni e difficoltà, in particolare legate all’accettazione della loro condizione di sopravvissute. Infatti, il migliore adattamento si riscontra nelle donne che hanno avuto peggiori condizioni in partenza: stadio del cancro avanzato, più giovani, con diagnosi ricevuta in età fertile. Pertanto, è possibile suggerire che queste condizioni critiche forzino queste donne ad affrontare apertamente il cancro e la loro situazione di sopravvissute al cancro, portandole ad “andare avanti” piuttosto che “tornare indietro”. Facendo riferimento alle connessioni tra variabili psicologiche e fisiologiche in queste donne, si è evidenziato che HRV è influenzata dalla presenza di figure significative ma, in particolare, è presumibile che sia influenzata da un’appropriata condivisione emotiva con queste figure. Si è anche evidenziato che poter continuare ad essere efficaci nel proprio contesto personale si riflette in un maggiore HRV, probabilmente in quanto permette di preservare il senso di sé, riducendo in questo modo lo stress derivante dall’esperienza cancro. Pertanto, HRV in queste donne risulta associato con un migliore adattamento psicologico. Inoltre, si è evidenziato che in queste donne la Qualità della Vita è profondamente influenzata dalla percezione dell’immagine corporea. Si tratta di un aspetto innovativo che è stato rilevato in questo campione e che, invece, nei precedenti studi non è stato indagato. In ultimo, la strategia di coping fatalismo sembra essere protettiva e sembra facilitare il processo di accettazione del cancro. Si spera sinceramente che le ricerche future possano superare i limiti del presente studio, come la scarsa numerosità e l’uso di strumenti di valutazione che, per alcuni aspetti come la scala Evitamento nel MAC, non centrano totalmente il target di indagine. Le traiettorie future di questo studio sono: aumentare il numero di osservazioni, reclutando donne in diversi centri specialistici in diverse zone d’Italia; utilizzare strumenti più specifici per valutare i costrutti in esame; valutare se un intervento di supporto centrato sul miglioramento di HRV (come HRV Biofeedback) può avere una ricaduta positiva sull’adattamento emotivo e la Qualità della Vita.

Epidemiology of ovarian cancer in Illinois /

"February 1996."

Ovulation and risk of epithelial ovarian cancer

Incessant ovulation is thought to be one of the primary causes of epithelial ovarian cancer. However, the effects of ovulation at different ages and of the various exposures or events that suppress ovulation have not been established. We used data from an Australian case-control study of 791 ovarian cancer cases and 853 controls to examine the effect of ovulation on ovarian cancer risk. The total number of lifetime ovulations was calculated using information provided in a monthly contraceptive/reproductive calendar, as well as incorporating other information such as average menstrual cycle length. An increase of I year's worth of ovulation was associated with a 6% increase in risk of ovarian cancer (95% confidence interval [CI] = 4-8%). Ovulations in the 20-29-year age group were associated with the greatest risk, with a 20% increase in risk associated with each year of ovulation during this age period (95% Cl = 13-26%). When the effects of different exposures that suppress ovulation were compared, there was an indication that some factors may have a greater effect than others. These findings support the theory that incessant ovulation is a major contributor to the occurrence of ovarian cancer and suggest that ovulations during the 20s may be those most associated with disease risk. (C) 2003 Wiley-Liss, Inc.

Characterisation of human kallikrein 6 protease M expression in ovarian cancer

Kallikrein 6 (hK6, also known as protease M/zyme/neurosin) is a member of the human kallikrein gene family. We have previously cloned the cDNA for this gene by differential display and shown the overexpression of the mRNA in breast and ovarian primary tumour tissues and cell lines. To thoroughly characterise the expression of this kallikrein in ovarian cancer, we have developed a novel monoclonal antibody specific to hK6 and employed it in immunohistochemistry with a wide range of ovarian tumour samples. The expression was found elevated in 67 of 80 cases of ovarian tumour samples and there was a significant difference in the expression levels between normal and benign ovarian tissues and the borderline and invasive tumours (P<0.001). There was no difference of expression level between different subtypes of tumours. More significantly, high level of kallikrein 6 expression was found in many early-stage and low-grade tumours, and elevated hK6 proteins were found in benign epithelia coexisting with borderline and invasive tissues, suggesting that overexpression of hK6 is an early phenomenon in the development of ovarian cancer. Quantitative real-time reverse transcription-polymerase chain reactions also showed elevated kallikrein 6 mRNA expression in ovarian tumours. Genomic Southern analysis of 19 ovarian tumour samples suggested that gene amplification is one mechanism for the overexpression of hK6 in ovarian cancer.

Alcohol, wine, and risk of epithelial ovarian cancer

Moderate alcohol intake can influence sex hormone levels and affect ovarian function as well as increasing breast cancer risk. This suggests that alcohol might also influence ovarian cancer risk. We have evaluated this among 696 Australian women with histologically confirmed epithelial ovarian cancer and 786 cancer-free control women, selected at random from the electoral roll. Sociodemographic information and a detailed reproductive history were collected in a face-to-face interview, and information about diet and alcohol consumption was obtained from a food frequency questionnaire. Logistic regression was used to calculate adjusted odds ratios (OR) and 95% confidence intervals (95% CI). Overall, 59% of women drank

Symptoms and diagnosis of borderline, early and advanced epithelial ovarian cancer

Objective. To document symptoms associated with borderline, early and advanced ovarian cancer and identify personal characteristics associated with early versus late diagnosis. Methods. Information concerning symptoms and diagnosis history was available from 811 women with ovarian cancer who took part in an Australian case–control study in the early 1990s. Women were classified into three groups for comparison based on their diagnosis: borderline, early (stage I–II) and advanced (stage III–IV) invasive cancer. Results. Sixteen percent of women with borderline tumors, 7% with early cancer and 4% with advanced cancer experienced no symptoms before diagnosis (P < 0.0001). Among women with symptoms, abdominal pain (44%) or swelling (39%) were most frequently reported; an abdominal mass (12%) and gynecological symptoms (12%) were less common. Compared to advanced stage cancer, women with early stage cancer were more likely to report an abdominal mass or urinary symptoms but less likely to report gastrointestinal problems or general malaise. General malaise and ‘other’ symptoms were least common in borderline disease. Older women, and those with higher parity or a family history of breast or ovarian cancer, were more likely to be diagnosed at an advanced stage of disease. Conclusions. Women who experience persistent or recurrent abdominal symptoms, particularly swelling and/or pain should be encouraged to seek medical attention and physicians should be alert to the possibility of ovarian cancer even in the absence of an abdominal mass. Further information about the prevalence of these symptoms in the general population is essential to assist physicians in patient management.

Height, age at menarche, and risk of epithelial ovarian cancer

Objective: We hypothesized that the hormonal changes of adolescence influence ovarian cancer risk particularly in younger women. We investigated this possibility by examining the relationship between ovarian cancer and adult height and age at menarche as both factors reflect pubertal hormonal levels. Methods: Participants were a population-based sample of women with incident ovarian cancer (n = 794) and control women randomly selected from the Australian Electoral Roll (n = 855). The women provided comprehensive reproductive and lifestyle data during a standard interview. Results: Although neither height nor age at menarche was significantly related to the risk of ovarian cancer overall, increasing height was associated with increasing risk of the subgroup of mucinous borderline ovarian cancer (odds ratio, 5.3; 95% confidence interval, 1.5-19.1 for women 175 cm compared with women < 160 cm, P-trend = 0.02). Similarly, later age at menarche was associated with increasing risk of mucinous borderline cancers (odds ratio, 3.8; 95% confidence interval, 1.3-11.4 for those with age at menarche >= 44 years compared with those < 12 years, P-trend = 0.003). Women with mucinous borderline cancers were significantly younger than the women diagnosed with invasive cancers (mean 44 versus 57 years; P < 0.0001). Conclusions: Development of mucinous borderline ovarian cancers, predominantly diagnosed in women ages under 50 years, seems to be associated with age at menarche and attained adult height. These results are consistent with our original hypothesis that pubertal levels of reproductive hormones and insulin-like growth factor-I influence ovarian cancer risk in younger women.

Double-strand break repair gene polymorphisms and risk of breast or ovarian cancer

Deficiencies in DNA repair have been hypothesized to increase cancer risk and excess cancer incidence is a feature of inherited diseases caused by defects in DNA damage recognition and repair. We investigated, using a case-control design, whether the double-strand break repair gene polymorphisms RAD51 5' untranslated region -135 G > C, XRCC2 R188H G > A, and XRCC3 T241M C > T were associated with risk of breast or ovarian cancer in Australian women. Sample sets included 1,456 breast cancer cases and 793 age-matched controls ages under 60 years of age, 549 incident ovarian cancer cases, and 335 controls of similar age distribution. For the total sample and the subsample of Caucasian women, there were no significant differences in genotype distribution between breast cancer cases and controls or between ovarian cancer cases and combined control groups. The crude odds ratios (OR) and 95% confidence intervals (95% CI) associated with the RAD51 GC/CC genotype frequency was OR, 1.10; 95% CI, 0.80-1.41 for breast cancer and OR, 1.22; 95% CI, 0.92-1.62 for ovarian cancer. Similarly, there were no increased risks associated with the XRCC2 GA/AA genotype (OR, 0.98; 95% CI, 0.76-1.26 for breast cancer and OR, 0.93; 95% CI, 0.69-1.25 for ovarian cancer) or the XRCC3 CT/TT genotype (OR, 0.92; 95% Cl, 0.77-1.10 for breast cancer and OR, 0.87; 95% CI, 0.71-1.08 for ovarian cancer). Results were little changed after adjustment for age and other measured risk factors. Although there was little statistical power to detect modest increases in risk for the homozygote variant genotypes, particularly for the rare RAD51 and XRCC2 variants, the data suggest that none of these variants play a major role in the etiology of breast or ovarian cancer.

RAD52 Y415X truncation polymorphism and epithelial ovarian cancer risk in Australian women

The RAD52 gene is involved in the homologous recombination repair pathway and is a plausible candidate ovarian cancer predisposition gene. We undertook a case-control comparison of 508 epithelial ovarian cancer cases (91 low malignant potential and 417 invasive) and 298 healthy controls to assess the RAD52 Y415X polymorphism as a risk factor for epithelial ovarian cancer in Australian women. Heterozygote frequencies of 2.6 and 4% were observed among cases and controls, respectively. The risk estimate was 0.55 (95%CI 0.24-1.24), suggesting that the RAD52 Y415X polymorphism is not associated with epithelial ovarian cancer in Australian women. (C) 2004 Elsevier Ireland Ltd. All rights reserved.

Over-expression of Eph and ephrin genes in advanced ovarian cancer: ephrin gene expression correlates with shortened survival

Background: Increased expression of Eph receptor tyrosine kinases and their ephrin ligands has been implicated in tumor progression in a number of malignancies. This report describes aberrant expression of these genes in ovarian cancer, the commonest cause of death amongst gynaecological malignancies. Methods: Eph and ephrin expression was determined using quantitative real time RT-PCR. Correlation of gene expression was measured using Spearman's rho statistic. Survival was analysed using log-rank analysis and ( was visualised by) Kaplan-Meier survival curves. Results: Greater than 10 fold over-expression of EphA1 and a more modest over-expression of EphA2 were observed in partially overlapping subsets of tumors. Over-expression of EphA1 strongly correlated ( r = 0.801; p < 0.01) with the high affinity ligand ephrin A1. A similar trend was observed between EphA2 and ephrin A1 ( r = 0.387; p = 0.06). A striking correlation of both ephrin A1 and ephrin A5 expression with poor survival ( r = - 0.470; p = 0.02 and r = - 0.562; p < 0.01) was observed. Intriguingly, there was no correlation between survival and other clinical parameters or Eph expression. Conclusion: These data imply that increased levels of ephrins A1 and A5 in the presence of high expression of Ephs A1 and A2 lead to a more aggressive tumor phenotype. The known functions of Eph/ephrin signalling in cell de-adhesion and movement may explain the observed correlation of ephrin expression with poor prognosis.

Benign epithelial ovarian tumours - cancer precursors or markers for ovarian cancer risk?

The natural history of the development of epithelial ovarian cancer remains obscure and no effective screening test exists. In several human malignancies progression from benign to invasive tumour occurs, but this sequence has not been established for epithelial ovarian cancer. We have reviewed epidemiological, histopathological and molecular studies of benign epithelial ovarian tumours to assess the evidence for and against such a progression in ovarian cancer. These data suggest that a diagnosis of a benign ovarian cyst or tumour is associated with an increased risk of ovarian cancer later in life. Current evidence also suggests that benign serous tumours can progress to low-grade serous cancer and that benign mucinous tumours can progress to mucinous cancer. The more common high-grade serous ovarian cancers are likely to arise de novo.

Expression profiling correlates with treatment response in women with advanced serous epithelial ovarian cancer

The majority of epithelial ovarian carcinomas are of serous subtype, with most women presenting at an advanced stage. Approximately 70% respond to initial chemotherapy but eventually relapse. We aimed to find markers of treatment response that might be suitable for routine use, using the gene expression profile of tumor tissue. Thirty one women with histologically-confirmed late-stage serous ovarian cancer were classified into 3 groups based on response to treatment (nonresponders, responders with relapse less than 12 months and responders with no relapse within 12 months). Gene expression profiles of these specimens were analyzed with respect to treatment response and survival (minimum 36 months follow-up). Patients' clinical features did not correlate with prognosis, or with specific gene expression patterns of their tumors. However women who did not respond to treatment could be distinguished from those who responded with no relapse within 12 months based on 34 gene transcripts (p < 0.02). Poor prognosis was associated with high expression of inhibitor of differentiation-2 (ID2) (p = 0.001). High expression of decorin (DCN) and ID2 together was strongly associated with reduced survival (p = 0.003), with an estimated 7-fold increased risk of dying (95% CI 1.9-29.6; 14 months survival) compared with low expression (44 months). Immunohistochemical analysis revealed both nuclear and cytoplasmic distribution of ID2 in ovarian tumors. High percentage of nuclear staining vas associated with poor survival, although not statistically significantly. In conclusion, elevated expression of ID2 and DCN was significantly associated with poor prognosis in a homogeneous group of ovarian cancer patients for whom survival could not be predicted from clinical factors. (c) 2006 Wiley-Liss, Inc.