SerpinB2 regulates stromal remodelling and local invasion in pancreatic cancer.

Pancreatic cancer has a devastating prognosis, with an overall 5-year survival rate of ~8%, restricted treatment options and characteristic molecular heterogeneity. SerpinB2 expression, particularly in the stromal compartment, is associated with reduced metastasis and prolonged survival in pancreatic ductal adenocarcinoma (PDAC) and our genomic analysis revealed that SERPINB2 is frequently deleted in PDAC. We show that SerpinB2 is required by stromal cells for normal collagen remodelling in vitro, regulating fibroblast interaction and engagement with collagen in the contracting matrix. In a pancreatic cancer allograft model, co-injection of PDAC cancer cells and SerpinB2-/- mouse embryonic fibroblasts (MEFs) resulted in increased tumour growth, aberrant remodelling of the extracellular matrix (ECM) and increased local invasion from the primary tumour. These tumours also displayed elevated proteolytic activity of the primary biochemical target of SerpinB2-urokinase plasminogen activator (uPA). In a large cohort of patients with resected PDAC, we show that increasing uPA mRNA expression was significantly associated with poorer survival following pancreatectomy. This study establishes a novel role for SerpinB2 in the stromal compartment in PDAC invasion through regulation of stromal remodelling and highlights the SerpinB2/uPA axis for further investigation as a potential therapeutic target in pancreatic cancer.

Gonadotropin-induced puberty does not impair reproductive performance of gilts over three parities.

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty-nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post-weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning-to-oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.

Adjuvant chemotherapy in elderly patients with pancreatic cancer.

BACKGROUND: Adjuvant chemotherapy improves survival for patients with resected pancreatic cancer. Elderly patients are under-represented in Phase III clinical trials, and as a consequence the efficacy of adjuvant therapy in older patients with pancreatic cancer is not clear. We aimed to assess the use and efficacy of adjuvant chemotherapy in older patients with pancreatic cancer. METHODS: We assessed a community cohort of 439 patients with a diagnosis of pancreatic ductal adenocarcinoma who underwent operative resection in centres associated with the Australian Pancreatic Cancer Genome Initiative. RESULTS: The median age of the cohort was 67 years. Overall only 47% of all patients received adjuvant therapy. Patients who received adjuvant chemotherapy were predominantly younger, had later stage disease, more lymph node involvement and more evidence of perineural invasion than the group that did not receive adjuvant treatment. Overall, adjuvant chemotherapy was associated with prolonged survival (median 22.1 vs 15.8 months; P<0.0001). Older patients (aged ≥70) were less likely to receive adjuvant chemotherapy (51.5% vs 29.8%; P<0.0001). Older patients had a particularly poor outcome when adjuvant therapy was not delivered (median survival=13.1 months; HR 1.89, 95% CI: 1.27-2.78, P=0.002). CONCLUSION: Patients aged ≥70 are less likely to receive adjuvant therapy although it is associated with improved outcome. Increased use of adjuvant therapy in older individuals is encouraged as they constitute a large proportion of patients with pancreatic cancer.

Identification of PUMA as an estrogen target gene that mediates the apoptotic response to tamoxifen in human breast cancer cells and predicts patient outcome and tamoxifen responsiveness in breast cancer.

Recognition of the pivotal role of estrogen in the aetiology of breast cancer has led to the development of antiestrogens (AE), such as tamoxifen (TAM) as effective therapies for the treatment and prevention of this disease. However, despite their widespread clinical efficacy, response to AEs is often short-lived, and acquired or innate therapeutic resistance remains a major obstacle in the successful treatment of breast cancer. Thus, delineating the intracellular pathways that mediate the cellular response to estrogen could potentially lead to new, more effective approaches to the treatment of breast cancer, particularly endocrine-resistant disease. Here, we have identified the BCL-2 homology 3 (BH3)-only, pro-apoptotic regulator, PUMA (p53 upregulated modulator of apoptosis) as an estrogen target gene that is acutely downregulated in response to estrogen in breast cancer cell lines, independently of their p53 status. PUMA is transcriptionally upregulated following treatment with TAM, and knock down of PUMA expression in these cells attenuates the apoptotic response to TAM. Furthermore, low PUMA expression in breast carcinomas is significantly associated with breast cancer-specific death (P=0.0014 and P=0.0115, for mRNA and protein, respectively), and worse outcome in TAM-treated patients (mRNA, P=1.49e-05). These findings suggest that the dysregulation of apoptotic signaling pathways such as those executed through PUMA, can significantly impact on both the progression and therapeutic responsiveness of breast cancer. Moreover, they provide a convincing rationale for exploring new therapeutic approaches involving endocrine and non-endocrine therapies that target apoptotic pathways as an effective strategy for tackling endocrine refractory disease.

BAG-1 predicts patient outcome and tamoxifen responsiveness in ER-positive invasive ductal carcinoma of the breast.

BAG-1 (bcl-2-associated athanogene) enhances oestrogen receptor (ER) function and may influence outcome and response to endocrine therapy in breast cancer. We determined relationships between BAG-1 expression, molecular phenotype, response to tamoxifen therapy and outcome in a cohort of breast cancer patients and its influence on tamoxifen sensitivity in MCF-7 breast cancer cells in vitro. Publically available gene expression data sets were analysed to identify relationships between BAG-1 mRNA expression and patient outcome. BAG-1 protein expression was assessed using immunohistochemistry in 292 patients with invasive ductal carcinoma and correlated with clinicopathological variables, therapeutic response and disease outcome. BAG-1-overexpressing MCF-7 cells were treated with antioestrogens to assess its effects on cell proliferation. Gene expression data demonstrated a consistent association between high BAG-1 mRNA and improved survival. In ER+ cancer (n=189), a high nuclear BAG-1 expression independently predicted improved outcome for local recurrence (P=0.0464), distant metastases (P=0.0435), death from breast cancer (P=0.009, hazards ratio 0.29, 95% CI: 0.114-0.735) and improved outcome in tamoxifen-treated patients (n=107; P=0.0191). BAG-1 overexpression in MCF-7 cells augmented antioestrogen-induced growth arrest. A high BAG-1 expression predicts improved patient outcome in ER+ breast carcinoma. This may reflect both a better definition of the hormone-responsive phenotype and a concurrent increased sensitivity to tamoxifen.

Different doses of porcine luteinizing hormone in precocious puberty induction in gilts.

The use of hormonal protocols in puberty induction and synchronization of oestrus has lead to an increase in the efficiency of replacement gilts. The aim of this study was to evaluate different doses of porcine LH in precocious puberty induction and oestrus synchronization in a homogeneous group of gilts. Sixty-seven homogeneous prepubertal gilts (Camborough 22) at 137 +/- 4 days of age and 87 +/- 7 kg were treated with three different hormonal protocols: T1--600 UI of equine chorionic gonadotrophin (eCG; Novormon) and after a 72-h period 5 mg of LH (Lutropin); T2--600 UI of eCG and a 72-h period 2.5 mg of LH; T3--600 UI of eCG and a 72-h period 1.25 mg of LH. The ovaries were examined at slaughter, on day 6 after the hormonal treatment. There were no statistical differences (p > 0.05) between the different LH doses in the percentage of the detected oestrus (T1 = 42.85%; T2 = 60.87%, T3 = 52.18%), oestrus duration (T1 = 41.44 +/- 16.30 h; T2 = 48.57 +/- 16.29 h, T3 = 39.33 +/- 11.42 h), number of corpora lutea (T1 = 9.61 +/- 5.43; T2 = 9.86 +/- 3.32, T3 = 8.13 +/- 5.52) and percentage of animals presenting ovarian cystic degeneration (T1 = 33.33%; T2 = 39.13%, T3 = 39.13%). The T2 (2.5 mg of LH) presented the lowest dispersion (p < 0.05) of the LH-ovulation interval (T1 = 37.17 +/- 4.07 h; T2 = 38.26 +/- 2.84 h; T3 = 36.25 +/- 5.69 h). The LH dose reduction to 2.50 and 1.25 mg presented equal results with the recommended dose of 5.0 mg, and could be used in the precocious induction of oestrus in gilts. The 2.5-mg LH dose showed the lowest dispersion of ovulation and it can be used in fixed-time artificial insemination programmes.

Relationship between prenatal survival rate at 70 days of gestation and morphometric parameters of vagina, uterus and placenta in gilts.

Swine uterine capacity affects litter size, and it could be used as a selection parameter of reproductive performance. Although there are some controversial results, evidences show that the catheter penetration length is positively correlated with litter size, and it could be used as a tool for predicting selection methods. The aim of this study was to determine whether there is any association between the prenatal survival rate and placental size at 70 days of gestation, the vaginal length [catheter penetration length during artificial insemination (AI)] and the uterine capacity in a homogeneous group of gilts. Sixty-six commercial-line gilts in pre-pubertal phase had their oestrus induced by hormonal treatment [600 UI of Equine Chorionic Gonadtrophin (eCG) i.m. and after a 72-h period 5 mg of luteinizing hormone (LH) i.m.], but only 40 gilts showed cyclicity after induction. The AI catheter penetration length was tested on these 40 gilts at the moment of AI using a calibrated AI catheter. Four gilts returned to oestrus and the other 36 were killed at around day 69 of pregnancy. The uterine length and weight showed a significant and positive correlation with the prenatal survival rate (p <0.05). The catheter penetration length was unable to predict the conceptus survival rate on 70 days of gestation; however, the uterine size influenced the survival rate positively. The mean placental area was positively correlated with the mean placental weight (p <0.0001), and both with the mean foetal weight (p <0.0001 and p <0.001, respectively). The analysis of the results obtained showed that neither did the catheter penetration length measurement during AI, nor the prenatal survival rate on day 70 of pregnancy predict the uterine capacity, but the uterine and placental size had a significant influence on the prenatal survival and foetus weight, respectively.