97 research outputs found

    Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells

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    Alternative splicing involves differential exon selection of a gene transcript to generate mRNA and protein isoforms with structural and functional diversity. Abnormal alternative splicing has been shown to be associated with malignant phenotypes of cancer cells, such as chemo-resistance and invasive activity. Screening small molecules and drugs for modulating RNA splicing in human hepatocellular carcinoma cell line Huh-7, we discovered that amiloride, distinct from four pH-affecting amiloride analogues, could “normalize” the splicing of BCL-X, HIPK3 and RON/MISTR1 transcripts. Our proteomic analyses of amiloride-treated cells detected hypo-phosphorylation of splicing factor SF2/ASF, and decreased levels of SRp20 and two un-identified SR proteins. We further observed decreased phosphorylation of AKT, ERK1/2 and PP1, and increased phosphorylation of p38 and JNK, suggesting that amiloride treatment down-regulates kinases and up-regulates phosphatases in the signal pathways known to affect splicing factor protein phosphorylation. These amiloride effects of “normalized” oncogenic RNA splicing and splicing factor hypo-phosphorylation were both abrogated by pre-treatment with a PP1 inhibitor. Global exon array of amiloride-treated Huh-7 cells detected splicing pattern changes involving 584 exons in 551 gene transcripts, many of which encode proteins playing key roles in ion transport, cellular matrix formation, cytoskeleton remodeling, and genome maintenance. Cellular functional analyses revealed subsequent invasion and migration defects, cell cycle disruption, cytokinesis impairment, and lethal DNA degradation in amiloride-treated Huh-7 cells. Other human solid tumor and leukemic cells, but not a few normal cells, showed similar amiloride-altered RNA splicing with devitalized consequence. This study thus provides mechanistic underpinnings for exploiting small molecule modulation of RNA splicing for cancer therapeutics

    Proceedings of the 20th Annual European Real Estate Society Conference - Vienna, Austria

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    Spatial complexity is not only one of the crucial sources of wayfinding problemwithin a shopping facility but also a significant determinant for space usage efficiency.The distribution of total floor area for a shopping mall is basically finding theoptimum solution for tenant location/allocation and pedestrian flow plans. Normally,with higher spatial complexity, it is easier for shoppers to get lost and generate highershopping costs; however, space usage flexibility could also be increased. Therefore, agood measurement for spatial complexity for shopping areas is needed. This papercompares several measurements for spatial complexity. The intention is to tackle thespatial complexity issue through four dimensions: forms and configurations,horizontal complexity, vertical movements and multiple-purposive users in mixed useenvironment. And the data is collected from the US, UK, Taiwan, Singapore, HongKong, Malaysia and Shanghai with more than 100 floorplans to explore the influentialfactors for spatial complexity within shopping centres. Using GIS, space syntax andother non-spatial techniques, this research suggests some interesting managementissues and enhances the understanding of spatial complexity within a shopping environment
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