β-EP和胃泌素在大鼠舌下腺的免疫组织化学定位

In order to study the distribution of β-EP and gastrin immunoreactive cells in the sublingual gland, we used the Advidin Biptin-Peroxidase Complex (ABC) immunohistochemical technique to detect the expression of β-EP and gastrin in rat sublingual glands. The results show that the epithelial cells of striated ducts and interlobular ducts appeared β-EP and gastrin positive in immunoreaction and the mucosus and serous acinar cells were immunoreactive negative. The poei-tive substances were distributed in the cytoplasm with negative nuclei. These results suggest that β-EP and gastrin are ex-pressed in the epithelial cells of ducts in the rat sublingual gland, which may play an important role in the secretion and functional regulation of the sublingual gland.     

Role of Lef1 in sustaining self-renewal in mouse embryonic stem cells

Embryonic stem cells (ESCs) can self-renew indefinitely while maintaining the ability to generate all three germ-layer derivatives.Despite the importance of ESCs in developmental biology and their potential impact on regenerative medicine,the molecular mechanisms controlling ESC behavior are incompletely understood.Previously,activation of the canonical Wnt signaling pathway has been shown to contribute to mouse ESC self-renewal.Here we report that ectopic expression of Lef1,a component of the Wnt signaling pathway,has a positive effect on the self-renewal of mouse ESCs.Lef1 up-regulates Oct4 promoter activity and physically interacts with Nanog,two key components of the ESC pluripotency machinery.Moreover,siRNA for Lef1 induced mouse ESC differentiation.Our results thus suggest that in response to Wnt signaling Lef1 binds to stabilized β-catenin and helps maintain the undifferentiated status of ESCs through modulation of Oct4 and Nanog.     

Silencing RhoA inhibits migration and invasion through Wnt/β-catenin pathway and growth through cell cycle regulation in human tongue cancer

Ras homolog gene family member A （RhoA） has been iden- tified as a critical regulator of tumor aggressive behavior. In this study, we assessed the role of RhoA in the mechan- isms underlying growth, migration, and invasion of squa- mous cell carcinoma of tongue （TSCC）. Stable RhoA knockdown of TSCC cell lines SCC-4 and CAL27 were achieved using Lentiviral transfection. The effects of RhoA depletion on cell migration, invasion, and cell proliferation were determined. The possible underlying mechanism of RhoA depletion on TSCC cell line was also evaluated by determining the expression of Galectin-3 （Gal-3）, β-catenin, and matrix metalloproteinase-9 （MMP-9） in vivo. Meanwhile, the underlying mechanism of TSCC growth was studied by analysis of cyclin D1/2, p21clel/WArl, and p27 kiap 1 protein levels. Immunohistochemical assess- ments were performed to further prove the alteration of Gal-3 and β-catenin expression. We found that, in mice injected with human TSCC cells in the tongue, RhoA levels were higher in primary tumors and metastasized lymph nodes compared with those in the normal tissues. Silencing of RhoA significantly reduced the tumor growth, decreased the levels of Gai-3, β-catenin, MMP-9, and cyclin D1/2, and increased the levels of p21 CIPI/WAFI and p27Kiap 1. In vitro, RhoA knockdown also led to inhibition of cell migration, in- vasion, and proliferation. Our data suggest that RhoA plays a significant role in TSCC progression by regulating cell migra- tion and invasion through Wnt/β-catenin signaling pathway and cell proliferation through cell cycle regulation, respecti- vely. RhoA might be a novel therapeutic target of TSCC.     

Knockdown of survivin expression by siRNAs enhances chemosensitivity of prostate cancer cells and attenuates its tumorigenicity

Survivin, a member of inhibitor of apoptosis family protein, has become an attractive therapeutic target in cancer due to its selective expression in tumor cells and its important roles for tumor cell viability. Here, we show that vector-based small interfering RNAs （siRNAs） silenced survivin expression in prostate cancer cells, resulting in significantly reduced cell proliferation and enhanced apoptosis, and increased the sensitivity of prostate cancer cells （PC-3） to the apoptosis-inducing agent, platinol. Furthermore, PC-3 cells transfected with the siRNA-expressing vector showed lower tumor formation in nude mice xenografts in vivo. These results demonstrated that inhibition of survivin expression by siRNA attenuated the malignant phenotypes of prostate cancer cells, and may provide a novel approach for gene therapy of androgen-independent prostate cancer.     

Eplerenone inhibits atrial fibrosis in mutant TGF-β1 transgenic mice

The purpose of the present study was to study the impacts of eplerenone(EPL), an antagonist of mineralocorticoid receptors(MR), on atrial fibrosis in a mouse model with selective fibrosis in the atrium, and to explore the possible mechanisms. Using mutant TGF-β1 transgenic(Tx) mice, we first demonstrated that EPL inhibited atrial fibrosis specifically and decreased macrophage accumulation in the atria of these mice. Results from immunohistochemistry and western blotting showed that EPL attenuated protein expression of fibrosis-related molecules such as connective tissue growth factor(CTGF) and fibronectin in the atria of Tx mice. In culture, EPL inhibited gene expression of fibrosis-related molecules such as fibronectin, α-SMA, and CTGF in TGF-β1-stimulated atrial fibroblasts. Finally, using a co-culture system, we showed that TGF-β1-stimulated atrial fibroblasts induced migration of macrophages and this was blocked by EPL. EPL also blocked TGF-β1-induced gene expression of intedeukin-6(IL-6) in atrial fibroblasts. Therefore, we conclude that EPL attenuated atrial fibrosis and macrophage infiltration in Tx mice. TGF-β1 and IL-6 were involved in the impacts of EPL on activation of atrial fibroblasts and interactions between fibroblasts and macrophages.     

GSK-3β as a driving force in ovarian cancer

Ovarian cancer is one of the most lethal malignancies in women. Identification of new therapeutic targets would provide opportunities for developing potentially more effective treatment regimes. In the July issue of Cell Research, Cao et al. reports that glycogen synthase kinase-3β （GSK-3β） plays an important role in positively regulating the proliferation of human ovarian cancer cells, and thus it may represent such a target [ 1 ]. GSK-3β is a serine/threonine kinase that is known to be involved in regulation of β-catenin signaling, where it participates in the formation of a multi-component destruction complex that promotes the phosphorylation and subsequent degradation of β-catenin. Given that overactive β-catenin signaling is involved in many forms of human cancer, this classic mode of GSK-3β action should qualify it as a ＂tumor suppressor＂. Intriguingly, however, two recent studies have implicated that GSK-3β may actually play a pro-tumor role in pancreatic and colorectal cancers [2, 3]. Since ovarian tumors often exhibit increased expression of GSK-3β, these recent findings prompted Cao et al. to examine the potential role of GSK-3β in ovarian cancer cells.     

Characterization of transgenic mice expressing EGFP under the control of the monkey 20α-hydroxysteroid dehydrogenase promoter

To directly assess the molecular function of the monkey 20α-hydroxysteroid dehydrogenase(20α-HSD) promoter, we generated transgenic mice(tg) expressing enhanced green fluorescent protein(EGFP) under control of this promoter. We demonstrated that prostaglandin F2α induced 20α-HSD promoter activity in CHO cells in a dose-dependent manner. Furthermore, forskolin treatment markedly reduced 20α-HSD promoter activity, and prolactin exhibited weak inhibitory activity. The transgenic mouse obtained one positive founder male. The transgene was propagated in 10 successive generations without any notable defects to the progeny. EGFP and 20α-HSD in the tg mice were colocalized in the luteal cells of the ovary during late pregnancy. Strong EGFP and 20α-HSD protein signals were also detected in the adult testis. Immunohistochemical analysis revealed high EGFP levels in the seminiferous epithelium, whereas 20α-HSD was expressed in the seminiferous tubules. Our data suggest that the ovaries in monkey and mouse exhibit similar expression patterns of 20α-HSD during pregnancy. However, the expression pattern of EGFP in tg mice testis slightly differed from that of the endogenous 20α-HSD. Further investigation is required to elucidate the functional mechanisms underlying regulation of the monkey 20α-HSD promoter in the tg mice.