Description: IntroductionTransforming growth factor beta (TGF?) plays a major role in the regulation of tumor initiation, progression, and metastasis. It is depended on the type II TGF? receptor (T?RII) for signaling. Previously, we have shown that deletion of T?RII in mammary epithelial of MMTV-PyMT mice results in shortened tumor latency and increased lung metastases. In MMTV-Neu mouse model, which is related to human epidermal growth factor 2 (HER2)?+?human breast cancer, active TGF? signaling increased the number of circulating tumor cells and metastases. Therefore, the loss-of-function experiments suppressed metastasis in Neu-induced tumors. In the current study, we describe a newly discovered connection between attenuated TGF? signaling and HER2 signaling in mammary tumor progression. Methods: All studies were performed on MMTV-Neu mice with and without dominant-negative T?RII (DNIIR) in mammary epithelium. Mammary tumors were analyzed by flow cytometry, immunohistochemistry, and immunofluorescence staining. The levels of secreted proteins were measured by enzyme-linked immunosorbent assay. Whole-lung mount staining was used to quantitate lung metastasis. The Cancer Genome Atlas (TCGA) datasets were used to determine the relevance of our findings to human breast cancer. Results: Attenuated TGF? signaling led to a delay of HER2+ tumor onset, but increased the number of metastases. The DNIIR tumors were characterized by increased vasculogenesis, vessel leakage, and increased expression of vascular endothelial growth factor (VEGF). During DNIIR tumor progression, both the levels of CXCL1/5 and the number of CD11b+Gr1+ cells and T cells decreased. Analysis of TCGA datasets demonstrated a significant negative correlation between TGFBR2 and VEGF genes expression. Higher VEGFA expression correlated with shorter distant metastasis-free survival only in HER2+ patients with no differences in HER2-, estrogen receptor +/- or progesterone receptor +/- breast cancer patients. Conclusion: Our studies provide insights into a novel mechanism by which epithelial TGF? signaling modulates the tumor microenvironment, and by which it is involved in lung metastasis in HER2+ breast cancer patients. The effects of pharmacological targeting of the TGF? pathway in vivo during tumor progression remain controversial. The targeting of TGF? signaling should be a viable option, but because VEGF has a protumorigenic effect on HER2+ tumors, the targeting of this protein could be considered when it is associated with attenuated TGF? signaling.

Description: Background: The study on the second generation bio-fuel is a hot area of current research of renewable energy. Among series of key points in this area, the role of beta-glucosidase in the degradation of intermediate gluco-oligosaccharides limits the rate of the complete saccharification of lignocellulose. Results: In this study, a new beta-glucosidase gene, unglu135B12, which was isolated from a metagenomic library of rumen of cattle feeding with Miscanthus sinensis by the function-based screening, encodes a 779 amino acid polypeptide that contains a catalytic domain belonging to glycoside hydrolase family 3 (GH3). It was recombinantly expressed, purified and biochemically characterized. The recombinant beta-glucosidase, unglu135B12, displayed optimum enzymatic activity at pH 5.0 at 38[degree sign]C, and showed the highest specific activity of 2.5 x 103 U/mg under this optimal condition to p-nitrophenyl-beta-D-glucopyranoside (pNPG), and its Km and Vmax values were 0.309 mmol/L and 7.292 mumol/min, respectively. In addition, the presence of Ca2+, K+, Na+ slightly improved beta-glucosidase activity of unglu135B12 by about 5%, while about 10 ~ 85% loss of beta-glucosidase activity was induced by addition of Mn2+, Fe3+, Zn2+, Cu2+. Interestingly, unglu135B12 was activated by glucose at the concentration lower than 40 mM. Conclusions: Our findings indicate that unglu135B12 is a new beta-glucosidase derived from rumen of cattle, and it might be a potent candidate for saccharification of lignocellulose in industrial application.