Antiangiogenic Activity of Acer tegmentosum Maxim Water Extract in Vitro and in Vivo

Angiogenesis, the formation of new blood vessels, is critical for tumor growth and metastasis. Notably, tumors themselves can lead to angiogenesis by inducing vascular endothelial growth factor (VEGF), which is one of the most potent angiogenic factors. Inhibition of angiogenesis is currently perceived as one of the most promising strategies for the blockage of tumor growth. In this study, we investigated the effects of Acer tegmentosum maxim water extract (ATME) on angiogenesis and its underlying signal mechanism. We studied the antiangiogenic activity of ATME by using human umbilical vein endothelial cells (HUVECs). ATME strongly inhibited VEGF-induced endothelial cell proliferation, migration, invasion, and tube formation, as well as vessel sprouting in a rat aortic ring sprouting assay. Moreover, we found that the p44/42 mitogen activated protein (MAP) kinase signaling pathway is involved in the inhibition of angiogenesis by ATME. Moreover, when we performed the in vivo matrigel plug assay, VEGF-induced angiogenesis was potently reduced when compared to that for the control group. Taken together, these results suggest that ATME exhibits potent antiangiogenic activity in vivo and in vitro and that these effects are regulated by the extracellular regulated kinase (ERK) pathway.

The influence of HGF on the expression of VEGF-C and its mechanism in human tongue squamous cell carcinoma Tca8113 cells / 实用口腔医学杂志

Objective:To investigate the influence of hepatocyte growth factor(HGF)on the expression of vascular endothelial growth factor C(VEGF-C)and the mechanism of HGF-induced VEGF-C expression in tongue squamous cell carcinoma Tca8113 cells.Methods:Tca8113 cells were cultured and exposed to HGF with various concentrations.The expression level of VEGF-C was assessed by ELISA.Signaling transduction inhibitors LY294002,U0126,SP600125,SB203580 was used to block PI3K/Akt,P44 /P22MAPK,JNK,P38MAPK signaling pathways,respectively.Then,the expression level of VEGF-C was detected by ELISA.Re-sults:The VEGF-C expression of Tca8113 cells increased at the beginning and decreased later with the increase of HGF concentra-tion.When the concentration of HGF was 40 ng/ml,VEGF-C expression level was the highest.Inhibitor LY294002 of PI3K/Akt and Inhibitor U0126 of P44 /P22MAPK significantly blocked the effects on HGF-induced VEGF-C up-regulation(P ＜0.01 ).Inhibitor SP600125 of JNK and inhibitor SB203580 of P38MAPK didn't interfere HGF-induced VEGF-C expression(P >0.05).Conclusion:HGF contributed to the expression of VEGF-C,PI3K/Akt and P44 /P22MAPK signaling pathways may be involved in HGF-induced VEGF-C up-regulation,and may play potential roles in lymphatic metastasis of oral squamous cell carcinoma.

Low salt induces the expression of cyclooxygenase-2 in a mouse macula densa cells through the activation of ERK、AP-1 pathways / 基础医学与临床

Objective To identify the effect of low salt medium on P44/42 kinase(ERK)、AP-1 in the expression of cyclooxygenase-2 (COX-2) in a mouse macula densa derived cell line (MMDD1 cells). Methods MMDD1 cells were transfected with luciferase reporter plasmid containing AP-1. Luciferase reporter assay were studied in normal salt(NS) and low salt(LS) medium; the expression of C-JUN、C-FOS were analyzed as well by Western Blotting. RT-PCR and Western Blotting were used to detect the mRNA and protein expression of COX-2. Expression of total and phosphorylated ERK was analyzed by Western Blotting in LS solution. The content of PGE2 in the supernatant was examined by ELISA. Results Phosphorylated ERK were markedly increased by LS treatment. The up-regulated COX-2 protein expression with LS was reduced with PD-98059 (ERK inhibitiors) at 20 ?mmol/L, PGE2 release was down-regulated as well. Luciferase activity of AP-1 was stimulated in LS, the up-regulated luciferase activity of AP-1 was attenuated by curcumin at 20 ?mmol/L (AP-1 inhibitor). The expression of C-JUN、C-FOS were increased as well. Low salt medium changed COX-2 mRNA abundance and protein expression was decreasedin medium with curcumin at 20 ?mmol/L.Conclusion Low salt induces the expression of COX-2 in MMDD1 cells through the activation of ERK、AP-1 pathways.

Correlation between Genotype and Phenotype in Korean patients with Spinal Muscular Atrophy / 대한소아신경학회지

BACKGROUND: Spinal muscular atrophy (SMA) is the second most common disease with autosomal recessive mode of inheritance in children and characterized by degeneration of anterior horn cells of the spinal cord resulting in weakness and wasting of voluntary muscles. This disease is caused by deletion of many candidate genes including SMN, p44, NAIP on chromosome 5q11.2-13.3. Although molecular characteristics of candidate genes were identified, genotype-phenotype correlation has not been clearly elucidated yet. Nevertheless, gene conversion, previously described as simply as gene deletion, appears to be very important mechanism as a molecular pathogenesis, and even makes more difficult to pursue the correlation. PURPOSE: This study was aimed to define the correlation between genotype and phenotype of SMA in Korean patients. The significance of SMN gene as well as NAIP gene, p44 gene in the progress of disease process and phenotypic correlation with gene conversion was evaluated. This study was also undertaken to determine the frequency of gene rearrangements in normal population. METHOD: Eight type I SMA patients and two type II SMA patients were studied. SMN, NAIP, and p44 gene deletion were analyzed by PCR amplification and restriction enzyme digestion with DraI, DdeI and AluI, respectively. p44 gene was also analyzed by SSCP. Gene conversion was defined by centromeric and telomeric SMN gene exon 7 to exon 8 PCR amplification followed by DdeI restiction enzyme digestion. RESULT: 1) Five of eight type I patients showed deletion of SMN, NAIP and p44 gene, while the rest of type 1 and all type II patients showed deletion of SMN gene only. 2) We examined SMN and NAIP gene deletion on 100 normal newborns, which showed the deletion of centromeric SMN gene in two newborns, the relative frequency of 2% in gene rearrangement. 3) There was one case of type I SMA showing deletion of telomeric SMN exon 7 but not SMN exon 8 suggestive of gene conversion occurred during the recombination as a molecular pathogenesis. CONCLUSION: The major deletion of SMA candidate genes, SMN, NAIP, and p44 gene appear to be involved in severe phenotype since these three candidate genes deletion were noted only in type 1 cases. However, SMN gene deletion only identified both in type 1 and type 2 explains that SMN gene may plan an major role in the pathogenesis of SMA and also suggests that other factors may be affecting the severity in spinal muscular atrophy. One patient with type I which showed the conversion of the centromeric SMN gene to the teleomeric gene strongly supports that SMN gene copy number may not be correlated with the severity in SMA. Our molecular findings suggest that phenotype is not clearly correlated with genotype. Prenatal screening should be carefully undertaken to interpretate because of high frequency of gene rearrangements in normal populations.