Effect of EGF and AREG Treatment During Porcine in vitro Maturation on in vitro Developmental Potential of Preimplantation Embryos
JOURNAL OF ANIMAL AND VETERINARY ADVANCES
Authors: Jeung, Se-Heon; Jeon, Yu-Byeol; Biswas, Dibyendu; Choi, Kyung-Chul; Jeung, Eui-Bae; Hyun, Sang-Hwan
Abstract
Porcine In Vitro Maturation (IVM) and In Vitro Fertilization (IVF) technique has been used and improved. However, polyspermic penetration, low rate of Blastocyst (BL) formation and poor quality of BLs are induced by imperfect nuclear and cytoplasmic maturation. It has been reported that Epidermal Growth Factor (EGF) is beneficial for oocyte maturation to improve the IVM system and Amphiregulin (AREG) is a growth factor containing an EGF-like domain Consequently, the present study was performed to investigate whether during porcine IVM system EGF and/or AREG supplementation is profitable for improving oocyte maturation and embryo development. In Experiment 1, oocytes were maturated with EGF (15 ng mL(-1)) or AREG (15 ng mL(-1)) or combination of both and after 44 h, oocytes were stained with Hoechst and metaphase II and development rate were evaluated. Without any growth factor served as a control. In Experiment 2, maturated oocytes from the previous groups were fertilized with proven sperm and penetration rate after 10 h of post-insemination was evaluated. In Experiment 3 in vitro matured porcine oocytes were stained with fluorescein isothiocyanate-labeled peanut agglutinin and the distribution of Cortical Granules (CGs) was evaluated by laser confocal microscopy. In Experiment 1, the oocytes maturation rate were significantly higher in EGF, AREG or combined group (70.9 +/- 15.5, 75.8 +/- 9.8 and 70.5 +/- 10.8%, respectively) compared to control (51.6 +/- 18.3%) but no significant difference among the treatments group. In Experiment 2, there was no significance of sperm penetration rate among the groups however, polyspermy and sperm number per oocytes were decreased significantly (p < 0.05) in combined group (26.7 +/- 12.0 and 1.35 +/- 0.2%, respectively) compared with control (43.5 +/- 13.1 and 1.63 +/- 0.3%, respectively). In Experiment 3, BL formation rate were significantly (p <= 0.05) higher in combined group (17.1 +/- 5.0%) compared with control and EGF group (9.4 +/- 3.2 and 13.1 +/- 4.6%, respectively) but the TE cell number and total cell number were significantly higher in (p < 6.05) combined group compared to control and other treatment group. In Experiment 3, the CG area was increased significantly (p < 0.05) in all treatment groups compared with control group (11.1 +/- 3.4, 9.7 +/- 3.0 and 10.4 +/- 2.8 vs. 4.1 +/- 1.8%, respectively). These results indicate that the addition of EGF and/or AREG to porcine IVM medium that enhances oocyte maturation and increases the total cell number. Combination of both growth factors also increases development rate and decreases polyspermy. Therefore, it is suggested that AREG can assist immature porcine oocytes to the metaphase II stage, enhance developmental potential in in vitro system and that there is due to synergistic effect of EGF and AREG during oocytes and embryonic development.
Expression of amphiregulin predicts poor outcome in patients with pancreatic ductal adenocarcinoma
DIAGNOSTIC PATHOLOGY
Authors: Wang, Li; Wu, Huanwen; Wang, Lili; Lu, Junliang; Duan, Huanli; Liu, Xuguang; Liang, Zhiyong
Abstract
Background: The validation of novel diagnostic, prognostic and predictive biomarkers in cancer is crucial for optimizing the choice and efficacy of personalized therapies. The aim of this study was to determine the epidermal growth factor receptor (EGFR), epidermal growth factor receptor variant III (EGFRvIII) and amphiregulin (AREG) protein expression levels and to evaluate the prognostic significance of EGFR, EGFRvIII and AREG in pancreatic ductal adenocarcinoma (PDAC). Methods: The EGFR, EGFRvIII and AREG protein levels in PDAC (n = 92) were examined by using immunohistochemistry. The associations between EGFRvIII expression, AREG expression, AREG/EGFR co-expression and clinicopathological factors were assessed, the correlation between AREG and EGFR expression was analyzed and the survival analyses were performed. Results: Among the lesions of PDAC, 12 (13 %) stained positive for EGFRvIII, 49 (53.3 %) stained positive for AREG and 22(23.9 %) stained double positive for AREG/EGFR. The relationships between each protein expression level and the clinicopathologic factors were examined, only AREG/EGFR co-expression was significantly related to tumor differentiation (P = 0.032). The correlation between AREG and EGFR expression was statistically insignificant (P = 0.709). Univariate survival analysis proved that high tumor-node-metastasis (TNM) stage, poor tumor differentiation and AREG expression were significant poor prognostic factors for disease-free survival (DFS) and overall survival (OS). By multivariate survival analysis, tumor differentiation was an independent poor prognostic factor for DFS (HR = 1.785, P < 0.05), whereas high TNM stage (HR = 2.25, P < 0.05), poor tumor differentiation (HR = 2.125, P < 0.01), positive resection margins (HR = 1.84, P < 0.05), and AREG expression (HR = 1.822, P < 0.05) were all independent poor prognostic factors for OS. Conclusions: In conclusion, our data indicate that AREG expression is an important prognostic biomarker in PDAC.