Biomedical subjects

Li-Wha Wu

Publications and source records attributed to Li-Wha Wu.

12 recordsLinked to original sources

PubMed2006

An osteoclastic protein-tyrosine phosphatase is a potential positive regulator of the c-Src protein-tyrosine kinase activity: a mediator of osteoclast activity.

This study tested the hypothesis that an osteoclastic protein-tyrosine phosphatase, PTP-oc, enhances osteoclast activity through c-Src activation. The effects of several resorption activators and inhibitors on PTP-oc expression, resorption activity, and c-Src activation were determined in rabbit osteoclasts. PTP-oc expression was assayed with immunoblots and semi-quantitative RT-PCR. Osteoclastic activity was determined by the resorption pit assay; and c-Src activation was monitored by P-tyr527 (PY527) dephosphorylation, and in vitro kinase assay. Treatment of osteoclasts with PTH, PGE2, 1,25(OH)2D3, IL-1, but not RANKL or IL-6, significantly stimulated resorption activity, increased PTP-oc mRNA and protein levels, and reduced c-Src PY527 level with corresponding activation of c-Src protein-tyrosine kinase activity. The PTP-oc antisense phosphorothioated oligo treatment blocked the basal and IL-1alpha-mediated, but not RANKL-mediated, resorption activity of isolated osteoclasts. The antisense oligo treatment also significantly reduced the average depth of resorption pits created by rabbit osteoclasts under basal conditions. Calcitonin and alendondrate, significantly reduced resorption activity and PTP-oc expression, and increased c-Src PY527 with corresponding reduction in its PTK activity. The cellular PTP-oc protein level correlated with the resorption activity. Among the various signaling proteins co-immunoprecipitated with PTP-oc, the resorption effectors caused corresponding changes in the tyrosyl phosphorylation level of only c-Src. The GST-PTP-oc fusion protein dephosphorylated PY-527-containing c-Src peptide in time- and dose-dependent manner in vitro. In summary, (1) PTP-oc is regulated in part at transcriptional level, (2) upregulation of PTP-oc in osteoclasts led to c-Src activation, and (3) PY527 of c-Src may be a cellular substrate of PTP-oc. These findings are consistent with the hypothesis that PTP-oc is a positive regulator of c-Src in osteoclasts.

Animals↗

PubMed2006

Cyclooxygenase-2 is involved in S100A2-mediated tumor suppression in squamous cell carcinoma.

S100A2 is considered a putative tumor suppressor due to its loss or down-regulation in several cancer types. However, no mechanism has been described for the tumor suppressor role of S100A2. In this study, ectopic expression of S100A2 in the human malignant squamous cell carcinoma cell line KB resulted in a significant inhibition of proliferation, migration, and invasion. Moreover, S100A2 significantly reduced the number of colonies (>or=0.5 mm) formed in semisolid agar and decreased tumor growth and burden in nude mice. cDNA microarray analysis was used to compare mRNA expression profiles of vector- and S100A2-expressing isogenic cells. Among the genes deregulated by S100A2, the expression of cyclooxygenase-2 (COX-2) mRNA was significantly suppressed by S100A2 (2.4-fold). Western blot analysis confirmed that S100A2 reduced the expression of COX-2 protein in stably and transiently transfected KB and RPMI-2650 cells. COX-2 is frequently overexpressed in various types of cancer and plays an important role in tumor progression. Partial restoration of COX-2 expression attenuated the antitumor effect of S100A2 both in vitro and in vivo. Although the interplay between S100A2 and COX-2 remains to be clarified, these findings first showed a potent antitumor role of S100A2 in squamous cell carcinoma partly via reduced expression of COX-2.

Animals↗

PubMed2006

Anti-angiogenesis mediated by angiostatin K1-3, K1-4 and K1-4.5. Involvement of p53, FasL, AKT and mRNA deregulation.

The molecular mechanism mediated by multiple forms of angiostatin via acting on proliferating vascular endothelium remains elusive. To address whether three forms of angiostatin, K1-3, K1-4 or K1-4.5, utilized similar or distinct pathways to mediate anti-angiogenesis, we adopted an adenoviral expression system to express secretable angiostatin molecules for CM collection. The anti-angiogenic activity of K1-3, K1-4 or K1-4.5 was confirmed by using proliferation, migration, tube formation and apoptotic assays of human endothelial cells. These angiostatin molecules at comparable expression level inhibited various in vitro angiogenesis assays with some variations. Furthermore, K1-3, K1-4 or K1-4.5 increased the expression of p53 protein and its downstream effectors, enhanced FasL-mediated signaling pathways, and decreased activation of AKT. At least three different receptors, Fas, integrin alpha(v)beta3 and ATP synthase, were involved in the anti-angiogenic action of angiostatin molecules. Besides, the expression of 189 genes at mRNA level was significantly altered by K1-3, K1-4 or K1-4.5. More than 70% of these genes participate in growth, inflammation, apoptosis, migration and extracellular matrix. Taken together, K1-3, K1-4 and K1-4.5, regardless of the number of kringles in the angiostatin molecules, mediated anti-angiogenesis via mostly similar pathways. We are the first to demonstrate the involvement of DAPK1 in the mediation of anti-angiogenesis by angiostatin.

Adenoviridae↗

PubMed2005

Amphiregulin as a tumor promoter for oral squamous cell carcinoma: involvement of cyclooxygenase 2.

Amphiregulin (AR), an epidermal growth factor (EGF)-like molecule, is a mitogen for keratinocytes. Squamous cell carcinoma (SCC) is a tumor derived from keratinoctyes. Expression of AR mRNA positively correlated with the clinical progression of 39 oral SCC. Oral SCC line, KB, was used as a model to study if increased expression of AR altered the biological behaviors of SCC cells. Exogenous AR dose-dependently enhanced the proliferation of KB cells expressing EGF receptor 1 (EGFR-1), a major receptor for AR, but little AR. Neutralizing anti-AR antibody significantly reversed the stimulatory effect of exogenous AR on KB cell proliferation. Ectopically expressed AR in stable clones manifested higher abilities to proliferate, migrate and invade Matrigel than vector control. Cyclooxygenase 2 (COX-2), but not metalloprotease 9 (MMP-9) mRNA, was increased in all the AR-expressing stable clones. In summary, AR behaves as a tumor promoter for oral SCC cells partly via increased expression of COX-2.

Amphiregulin↗

PubMed2005

S100A2, a potential marker for early recurrence in early-stage oral cancer.

Early-stage oral cancer patients may have distinct clinical outcomes and respond differently to the same treatment. Up to now, there is still no individual marker to identify such patients with poor outcome. Down-regulation of a tumor suppressor gene, S100A2, in oral cancer cells was identified by mRNA profiling analysis then confirmed by RT-PCR and Southern blotting. The expression of nuclear S100A2 protein examined by immunohistochemistry was not significantly associated with any patient characteristic among the 70 early-stage oral squamous cell carcinoma (SCC) patients. Intriguingly, the loss of nuclear S100A2 positivity was significantly associated with shorter disease-free survival (p=0.019) while having no effect on the overall survival of these patients. Cox regression analysis with backward elimination identified S100A2 (p=0.006), tobacco smoking (p=0.013), and betel quid chewing (p=0.019) as independent predictors of disease-free survival. This is a first study to demonstrate that loss of nuclear S100A2 may serve as an independent prognostic marker for early-stage oral cancer patients at high risk of recurrence. A more aggressive treatment modality and intensive follow-up may be recommended for the patients with reduced expression of S100A2 in tumor cell nuclei.

Adult↗

PubMed2004

Participation of cyclin D1 deregulation in TNP-470-mediated cytostatic effect: involvement of senescence.

Inhibition of angiogenesis is becoming one promising, alternative approach to stop tumor from growth and spreading to distant organs. TNP-470, an analog of fumagillin, possesses potent anti-angiogenic effects with minimal toxicity in animal tumor models and is now in the phase III of human cancer trial. Although TNP-470 induced endothelial cell cycle arrest at G1 phase via p53 and p21(Cip1), the underlying mechanism of the cytostatic effect of TNP-470 on endothelial cells remains limited. We have found that TNP-470 did not only induce p53 and p21(Cip1) but also cyclin D1 in the basic fibroblast growth factors (bFGF)-treated endothelial cells. The TNP-470-mediated increase of cyclin D1 protein was due to the enhanced expression of mRNA. The induced cyclin D1 formed a complex with cyclin-dependent kinase4 (CDK4) and p21(Cip1). The ability of cyclin D1-associated CDK4 to phosphorylate retinoblastoma (Rb) protein was, however, reduced in the same cells. TNP-470 also significantly increased senescence-associated-beta-galactosidase activity (SA-gal), hallmark of cells undergoing senescence. Interestingly, the effect of increased cyclin D1 protein mimicked by overexpression of cyclin D1 increased the sensitivity of human umbilical vein endothelial cells (HUVECs) to TNP-470. In summary, the cytostatic effect of TNP-470 on endothelial cells is in part mediated by induction of senescence and cyclin D1 is a key molecule participating in this event.

Antibiotics, Antineoplastic↗

PubMed2004

The mRNA profile of genes in betel quid chewing oral cancer patients.

Oral cancer is one of the most common types of human cancer in the world. Although the risk factors for oral cancer are well-recognized in different countries, the molecular mechanism responsible for this malignancy remains elusive particularly in the countries where betel quid chewing is prevalent. The cDNA microarray analysis was used to analyse the mRNA expression patterns of 1177 genes in ten oral cancer patients with betel quid chewing history. Eighty-four genes involving cell adhesion, cell shape, growth, apoptosis, angiogenesis, metastasis, and metabolism were deregulated. Although the expression profile of these genes was shared by certain clinical patients, there was no significant association of the expression profile with clinical staging. Functional implication of four validated genes including caspase-1, STAT-1, COX-2 and pleiotrophin was discussed. This study provides pilot data for understanding the pathogenesis of oral cancer in countries like Taiwan where betel quid chewing is prevalent.

Adult↗

PubMed2004

Polyacetylenes function as anti-angiogenic agents.

PURPOSE: To investigate the antiangiogenic effects of plant extracts and polyacetylenes isolated from Bidens pilosa Linn., which is a popular nutraceutical herbal tea and folk medicine in anti-inflammatory, antitumor, and other medications worldwide. METHODS: Anti-cell proliferation, anti-tube formation, and cell migration assays were used for the valuation of bioactivities of target plant extracts and phytocompounds against angiogenesis. Bioactivity-guided fractionation, HPLC, and various spectral analyses were used to identify active fraction and phytocompounds for anti-angiogenesis. RESULTS: We show that an ethyl acetate (EA) fraction of B. pilosa exhibited significant anti-cell proliferation and anti-tube formation activities against human umbilical vein endothelium cells (HUVEC). Bioassay-guided fractionation led to isolation of one new and one known polyacetylenes, 1,2-dihydroxytrideca-5,7,9,11-tetrayne (1) and 1,3-dihydroxy-6(E)- tetradecene-8,10,12-triyne (2), respectively, from the EA fraction. Compounds 1 and 2 manifested highly specific and significant activities against HUVEC proliferation with IC50 values of 2.5 and 0.375 microg/ml, respectively, however, compound 1 had a more potent effect on preventing tube formation of HUVEC than compound 2 at a dose of 2.5 microg/ml. Western blot analysis showed that both compounds upregulated p27(Kip) or p21(Cip1), cyclin-dependent kinase inhibitors, in HUVEC. CONCLUSIONS: This is the first report to demonstrate that polyacetylenes possess significant anti-angiogenic activities and the ability to regulate the expression of cell cycle mediators, for example, p27(Kip1), p21(Cip1), or cyclin E.

Acetylene↗

PubMed2003

Angiostatin antagonizes the action of VEGF-A in human endothelial cells via two distinct pathways.

Angiostatin consisting of the first four-kringle domains of the plasminogen potently inhibits angiogenesis in vitro and in vivo. However, the molecular mechanism of action whereby angiostatin mediates its inhibitory effect on proliferating endothelial cells remains elusive. We therefore used the proliferating cultured human umbilical vein endothelial cells (HUVECs) promoted by vascular endothelial growth factor A to identify the endogenous signaling elements that mediate the antiangiogenic effect of angiostatin. Treatment of HUVEC with angiostatin at a concentration known to inhibit cell proliferation and induce apoptosis resulted in induction of p53-, Bax-, and tBid-mediated release of cytochrome c into the cytosol. In addition, angiostatin also activated the Fas-mediated apoptotic pathway in part via up-regulation of FasL mRNA, down-regulation of c-Flip, and activation of caspase 3. These results suggest that the anti-angiogenic action of angiostatin is likely mediated by two distinct signaling pathways, one intrinsic mediated by p53 while the other extrinsic involved in FasL engagement and mitochondria dysfunction.

Angiostatins↗

PubMed2002

Two UVC-induced stress response pathways in HeLa cells identified by cDNA microarray.

Environmental toxins induce multiple effects in vivo, involving various molecular pathways. The ultraviolet C (UVC, 254 nm) component of sunlight can cause strong cytotoxic and genotoxic effects. In this study, UVC-induced stress response factors were analyzed by cDNA microarray, using the Millennium(R) Nylon membrane chip system. HeLa cells were irradiated with 30 Joule/m(2)/sec UVC, incubated for 30 or 60 minutes and then subjected to the analysis. Multiple chips were used for each experimental condition so that the data could be analyzed statistically. Principal component analysis (PCA) was used to identify groups of genes whose expression changed in a similar manner with time post-UVC irradiation. Three major factors were identified, depending on the directionality of expression changes in each gene. The factor loadings in all three identified gene groups were high, indicating that genes within each group were highly correlated. Two factors exhibited significantly changed expression patterns after 30 minutes of incubation but in the opposite direction. This indicates that the "immediate early" UVC-induced stress response was elicited by two major pathways. Interestingly, expression of the genomic damage-inducible GADD genes, as well as p53, was initially decreased, unlike the "immediate early" genes Fos/Jun and Egr-1, which were strongly increased after 30 minutes of incubation. The results indicate that PCA used in the analysis of pre-hypothesized, functionally related genes can identify the potential subpathways in a group. This method provides a novel approach for identifying functionally-related genes in microarray studies.

Enzymes↗

PubMed2002

Molecular characterization of angiogenic properties of human oral squamous cell carcinoma cells.

Little is known about the specificity of angiogenic properties of oral cancer cells and the possible mechanisms. Stimulatory effects on proliferation and migration of human umbilical vein endothelial cells (HUVEC) characterized the angiogenic properties of oral cancer cells but not normal oral keratinocytes (NOK). ELISA found the presence of vascular endothelial growth factors (VEGF) both in the tested oral cancer cells and NOK. Attenuation of the proangiogenic effects by neutralizing VEGF antibodies suggests VEGF play a key role in the acquisition of the angiogenic phenotype in oral cancer cells. Western blotting of p53 and murine double mutant 2 (Mdm2) together with p53 DNA sequencing analysis indicate that p53 function loss by mutation or overexpression of Mdm2 occurred in all tested oral cancer cells regardless of their etiology. In summary, the angiogenic property of oral cancer cells is mediated by many factors in addition to VEGF and the functional status of p53.

Carcinoma, Squamous Cell↗

PubMedDate not supplied

Thrombospondin-1 acts as a fence to inhibit angiogenesis that occurs during cervical carcinogenesis.

PURPOSE: The acquisition of an angiogenic phenotype (angiogenic switch) is essential for cervical carcinogenesis. This study was aimed to examine the spatial and temporal relationship of thrombospondin-1 (TSP-1) expression in patients with precursor lesions and squamous cell carcinoma of uterine cervix and to correlate its expression with tumor angiogenesis. PATIENTS AND METHODS: TSP-1 expression and microvessel density were assessed by immunohistochemistry in samples obtained from patients with pathological diagnoses of cervical intraepithelial neoplasm I, carcinoma in situ, invasive squamous cell carcinoma (SCC), and benign disease (N = 12 from each group). Two representative blocks that contained serial changes of cervical lesions from these 48 subjects were examined, and the pathological findings were categorized into the four groups of (1) normal cervical epithelia, (2) low-grade squamous intraepithelial lesions (LSILs), (3) high-grade SILs (HSILs), and (4) SCC. RESULTS: A total of 120 foci with various cervical lesions from 98 slides were examined and classified into normal (48), LSIL (36), HSIL (24), and SCC epithelium (12). Immunohistochemical studies showed that TSP-1 was mainly localized at the basal epithelial cells, and we named it as the "TSP-1 fence." The mean microvessel density counts and TSP-1 scores for normal, LSIL, HSIL, and SCC epithelium were 7.3 +/- 2.9, 9.9 +/- 3.4, 17.7 +/- 5.1, and 22.8 +/- 8.6, and 3.8 +/- 0.4, 3.8 +/- 0.4, 1.8 +/- 0.4, and 1.5 +/- 0.5, respectively. The TSP-1 intensities were significantly higher and the MVD counts lower in the groups of normal and LSIL epithelium than in those with HSIL and SCC epithelium. In addition, microvessel density count was negatively associated with the intensity of TSP-1. DISCUSSION: Our results indicate that the disruption of TSP-1 fence and the switch to angiogenic phenotype occurred during the transition from LSIL into HSIL. This concordance suggests that TSP-1 plays a role in the regulation of angiogenic switch. We conclude thatthe onset of angiogenesis is an early event in cervical carcinogenesis due, in part, to the down-regulation of TSP-1 by the dysplastic epithelium.

Angiogenesis Inhibitors↗