Rho GTPase-activating protein deleted in liver cancer suppresses cell proliferation and invasion in hepatocellular carcinoma

Deleted in liver cancer (DLC1) is a candidate tumor suppressor gene recently isolated from human hepatocellular carcinoma. Structurally, DLC1 protein contains a conserved GTPase-activating protein for Rho family protein (RhoGAP) domain, which has been thought to regulate the activity of Rho family proteins. Previous studies indicated that DLC1 was frequently inactivated in cancer cells. In the present study, we aimed to characterize the tumor suppressor roles of DLC1 in hepatocellular carcinoma. We showed that DLC1 significantly inhibited cell proliferation, anchorage-independent growth, and in vivo tumorigenicity when stably expressed in hepatocellular carcinoma cells. Moreover, DLC1 expression greatly reduced the motility and invasiveness of hepatocellular carcinoma cells. With RhoGAP-deficient DLC1 mutant (DLC1-K714E), we showed that the RhoGAP activity was essential for DLC1-mediated tumor suppressor function. Furthermore, the 292- to 648-amino acid region and the steroidogenic acute regulatory related lipid transfer domain played an auxiliary role to RhoGAP and tumor suppressor function of DLC1. Taken together, our findings showed that DLC1 functions as a tumor suppressor in hepatocellular carcinoma and provide the first evidence to support the hypothesis that DLC1 suppresses cancer cell growth by negatively regulating the activity of Rho proteins. ©2005 American Association for Cancer Research.postprin

Structure and Photoluminescent Properties of ZnO Encapsulated in Mesoporous Silica SBA-15 Fabricated by Two-Solvent Strategy

The two-solvent method was employed to prepare ZnO encapsulated in mesoporous silica (ZnO/SBA-15). The prepared ZnO/SBA-15 samples have been studied by X-ray diffraction, transmission electron microscope, X-ray photoelectron spectroscopy, nitrogen adsorption–desorption isotherm, and photoluminescence spectroscopy. The ZnO/SBA-15 nanocomposite has the ordered hexagonal mesostructure of SBA-15. ZnO clusters of a high loading are distributed in the channels of SBA-15. Photoluminescence spectra show the UV emission band around 368 nm, the violet emission around 420 nm, and the blue emission around 457 nm. The UV emission is attributed to band-edge emission of ZnO. The violet emission results from the oxygen vacancies on the ZnO–SiO2interface traps. The blue emission is from the oxygen vacancies or interstitial zinc ions of ZnO. The UV emission and blue emission show a blue-shift phenomenon due to quantum-confinement-induced energy gap enhancement of ZnO clusters. The ZnO clusters encapsulated in SBA-15 can be used as light-emitting diodes and ultraviolet nanolasers

Role of hypoxia-induced upregulation of caveolin-1 in hepatocellular carcinoma cell migration and invasion

Poster Session: Cellular and Molecular Biology 30 Hypoxia: abstract no. 2049Intratumoral hypoxia occurs when cells in the inner core of solid tumor mass become deprived of oxygen. It correlates with an increased risk to develop metastasis. However, the molecular basis about the adaptation and survival of tumor cells in hypoxic environment remains obscure. Our recent data revealed that Cav1, a component of focal adhesion, is dramatically increased in metastatic hepatocellular carcinoma (HCC) cell lines and tissues, suggesting a role of Cav1 in HCC metastasis. In this study, we further investigated the potential role of Cav1 in HCC metastasis under hypoxia. Our data revealed that Cav1 mRNA and protein expressions were enhanced in various HCC cells lines when placed in hypoxic chamber or treated with hypoxic mimetic drug. Such upregulation of Cav1 was abolished with the addition of HIF1α inhibitor or depletion of HIF1α by siRNA. This suggests that hypoxia-induced Cav1 expression is HIF1α-dependent. Our findings also showed that HCC cells possessed enhanced motility under hypoxia. However, knockdown of Cav1 in HCC cells attenuated the capacity of cells to migrate and invade. Collectively, our study suggests that the increased motility of HCC cells conferred by the hypoxia-induced Cav1 expression is an adaptive mechanism of cells to survive under hypoxic stress.link_to_OA_fulltex

Cloning and characterization of the promoter region of the mouse frizzled-related protein 4 gene

Frizzled-related protein (Frp) is a newly identified family of secreted proteins involved in the Wnt signaling pathway. To date, little is known about the underlying mechanisms regulating Frp expression. In this study the promoter region of mouse frizzled related protein 4 (sFrp4) gene was cloned, sequenced, and analyzed using transient reporter assays along with sitedirected mutagenesis. Two clusters of cisacting elements, STAT3/Lyf-1/MZF1 (site 1) and C/EBP[beta]/ GATA-1/CREB (site 2) located in the promoter region from 238 to 144 were found to be essential for the promoter activity of sFrp4. In addition to sites 1 and 2, putative transcriptional factor binding sites for TFIID, SP1/GC and ATF/CREB exhibited positive, while the site for NRSE exhibited negative regulatory functions, as determined by the alkaline phosphatase activities of the reporter assay. We also demonstrate that the ATF/CREB site may cooperatively interact with the NRSFlike element in regulating sFrp4 promoter activity. The data of our study, which is the first promoter analysis of mouse Frp genes, provide the basis for understanding the functions and the regulation of Frp and its role in regulating Wnt signals

c-Met overexpression contributes to the acquired apoptosis resistance of non-adherent ovarian cancer cells through a cross-talk mediated by phosphatidylinositol 3-kinase and extracellular signal-regulated kinase 1/2

Poster Session 29 - Receptor Signaling in Endocrine Related Cancers: abstract no. 1722Ovarian cancer is the most lethal gynecological cancer mainly due to widespread peritoneal dissemination and malignant ascites. Key to this is the capacity of tumor cells to escape suspension-induced apoptosis (anoikis), which also underlies their resistance to chemotherapy. Here we utilized a non-adherent cell culture model to investigate molecular mechanisms of apoptotic resistance of ovarian cancer cells that may mimic the chemoresistance found in solid tumors. We found that ovarian cancer cells acquired a remarkable resistance to anoikis and apoptosis induced by exposure to clinically relevant doses of two front-line chemotherapeutic drugs cisplatin and paclitaxel when grown in three-dimensional (3D) than monolayer cultures. Inhibition of the hepatocyte growth factor (HGF) receptor c-Met, which is frequently overexpressed in ovarian cancer, by a specific inhibitor or small interfering RNA blocked the acquired anoikis resistance and restored chemosensitivity in 3D, not in 2D cultures. These effects were found to be dependent on both phosphatidylinositol 3-kinase (PI3K)/Akt and extracellular-signal regulated kinase (ERK) 1/2 signaling pathways. Inhibitors of PI3K/Akt abrogated ERK1/2 activation and its associated anoikis resistance in response to HGF, suggesting a signaling relay between these two pathways. Furthermore, we identified, as a mechanism of this cross-talk, a central role of Ras. Interestingly, Ras did not lie upstream of PI3K/Akt, whereas PI3K/Akt signaling to ERK1/2 involved Ras. These findings shed new light on the apoptotic resistance mechanism of non-adherent ovarian cancer ascites cells.link_to_OA_fulltextThe 101st Annual Meeting of the American Association for Cancer Research (AACR 2010), Washington D.C., 17-21 April 2010. In AACR Meeting Abstracts, 201

c-Met overexpression contributes to the acquired apoptotic resistance of nonadherent ovarian cancer cells through a cross talk mediated by phosphatidylinositol 3-kinase and extracellular signal-regulated kinase 1/2

Ovarian cancer is the most lethal gynecologic cancer mainly because of widespread peritoneal dissemination and malignant ascites. Key to this is the capacity of tumor cells to escape suspension-induced apoptosis (anoikis), which also underlies their resistance to chemotherapy. Here, we used a nonadherent cell culture model to investigate the molecular mechanisms of apoptotic resistance of ovarian cancer cells that may mimic the chemoresistance found in solid tumors. We found that ovarian cancer cells acquired a remarkable resistance to anoikis and apoptosis induced by exposure to clinically relevant doses of two front-line chemotherapeutic drugs cisplatin and paclitaxel when grown in three-dimensional than monolayer cultures. Inhibition of the hepatocyte growth factor (HGF) receptor c-Met, which is frequently overexpressed in ovarian cancer, by a specific inhibitor or small interfering RNA blocked the acquired anoikis resistance and restored chemosensitivity in three-dimensional not in two-dimensional cultures. These effects were found to be dependent on both phosphatidylinositol 3-kinase (PI3K)/Akt and extracellular signal-regulated kinase (ERK) 1/2 signaling pathways. Inhibitors of PI3K/Akt abrogated ERK1/2 activation and its associated anoikis resistance in response to HGF, suggesting a signaling relay between these two pathways. Furthermore, we identified a central role of Ras as a mechanism of this cross talk. Interestingly, Ras did not lie upstream of PI3K/Akt, whereas PI3K/Akt signaling to ERK1/2 involved Ras. These findings shed new light on the apoptotic resistance mechanism of nonadherent ovarian cancer ascites cells and may have important clinical implications. Copyright © 2010 Neoplasia Press, Inc. All rights reserved.link_to_OA_fulltex

Transcriptional activity of the promoter region of rat frizzled-related protein gene

Frizzled-related protein (Frp) is a new family of secreted proteins involved in tumorigenesis and Wnt-signaling pathway. Previous study has shown that rat Frp (rFrp) gene was found to be differentially expressed in Rat 6 fibroblast cell line overexpressing p53(val135) (R6\#13-8). The rFrp gene was otherwise silent in normal parental Rat 6 cells. To elucidate the molecular basis of the transcriptional activation of rFrp, we have isolated and analyzed a 2-kilobase pair promoter region of the rFrp gene. Mapping of transcription initiation sites of rFrp showed the existence of multiple initiation sites. Transfection studies of serial deletion constructs in both Rat 6 and CHOK1 cell lines revealed that the region from -202 to -144 contains cis-acting elements essential for the efficient transcription of rFrp. This work provides a transcriptional regulation basis for Frp and gives insight into its implication in tumorigenesis. (C) 2001 Academic Press