L to mount a more efficient UPR response and maintain extracellular

L to mount a more efficient UPR response and maintain extracellular matrix production, which may contribute to metastasis and cell survival. Analysis of The Cancer Genome Atlas (cancergenome.nih.gov/) glioma expression database [40], as well as the GBMBase (http:// www.gbmbase.org) which focuses on glioblastoma multiformeresearch, indicates that OASIS and various ER Epigenetics stress response genes are changed in gliomas relative to control tissue (Supplemental data Table S1 and Figure S1). Although OASIS expression can be both increased and decreased in primary human tumors its expression is increased in the majority of glioblastoma subcutaneous xenograph tumors (Supplemental data Table S1 and Figure S1). In future studies it will be important to examine OASIS protein expression and activation in primary human gliomas, as well as examining if targeting ER stress and OASIS may present new strategies to reduce glioma cell growth or infiltration using in vivo models. In summary, we identified that the ER stress sensor OASIS is a glycoprotein that is differentially expressed in human glioma cell lines. OASIS protein is induced by ER stress and appears to contribute to both maximal induction of the UPR (chaperone capacity), as well as maintaining extracellular matrix (CSPG) protein expression in glioma lines that express this protein. Because of these effects, we hypothesize that gliomas that express OASIS may be better off under hypoxic conditions in vivo and this may contribute to more resistant and invasive cancers.OASIS in Human Glioma CellsFigure 7. OASIS knock-down does not induce U373 cell apoptosis. (A) U373, A172 and U87 cells were treated with 1 mM TG for the times indicated prior to lysis and immunoblotting for cleaved caspase 3. Rat pancreatic INS-1 insulinoma cells treated with staurosporine were used as a positive control for Epigenetics detection of cleaved caspase 3. (B) U373 cells transfected with 100 nM control (GFP) or OASIS siRNAs for 7 days, then treated or not with 1 mM TG for an additional 48 h. The cells were lysed and immunoblotted for the indicated proteins. Pancreatic INS-1 cells treated with staurosporine (ST, 3 h or 24 h) were used as a positive control for detection of cleaved caspase 3. (C) U87 cells were treated with siRNAs as in (B), then with TG for 16 or 48 h and total protein per well was measured in cell lysates. N = 3 independent experiments. Bars are SEM. doi:10.1371/journal.pone.0054060.gSupporting InformationFigure S1 Expression of selected ER stress and ECM genes inAcknowledgmentsWe thank Children, comments fellowship 1516647 Toronto). Dr. James Rutka and Dr. Christian Smith (Hospital for Sick Toronto) for providing the human glioma cell lines and on the manuscript. RNV was funded by a post-doctoral from the Banting and Best Diabetes Centre (University ofhuman glioblastoma multiforme (GBM) tumors (DOCX)Table S1 Expression of selected ER stress and ECM genes inhuman glioblastoma multiforme (GBM) tumors (DOCX)Author ContributionsConceived and designed the experiments: RV AV. Performed the experiments: RV LZ. Analyzed the data: RV LZ AV. Wrote the paper: RV AV.OASIS in Human Glioma Cells
Dynamins are large GTPases involved in a wide range of cell and organelle fission events. The dynamin superfamily is made up of classical dynamins and dynamin-like proteins. Classical dynamins are critical components of clathrin-mediated endocytosis, where they contribute to the release of newly formed endosomes [1,2,3]. In addition to this well-char.L to mount a more efficient UPR response and maintain extracellular matrix production, which may contribute to metastasis and cell survival. Analysis of The Cancer Genome Atlas (cancergenome.nih.gov/) glioma expression database [40], as well as the GBMBase (http:// www.gbmbase.org) which focuses on glioblastoma multiformeresearch, indicates that OASIS and various ER stress response genes are changed in gliomas relative to control tissue (Supplemental data Table S1 and Figure S1). Although OASIS expression can be both increased and decreased in primary human tumors its expression is increased in the majority of glioblastoma subcutaneous xenograph tumors (Supplemental data Table S1 and Figure S1). In future studies it will be important to examine OASIS protein expression and activation in primary human gliomas, as well as examining if targeting ER stress and OASIS may present new strategies to reduce glioma cell growth or infiltration using in vivo models. In summary, we identified that the ER stress sensor OASIS is a glycoprotein that is differentially expressed in human glioma cell lines. OASIS protein is induced by ER stress and appears to contribute to both maximal induction of the UPR (chaperone capacity), as well as maintaining extracellular matrix (CSPG) protein expression in glioma lines that express this protein. Because of these effects, we hypothesize that gliomas that express OASIS may be better off under hypoxic conditions in vivo and this may contribute to more resistant and invasive cancers.OASIS in Human Glioma CellsFigure 7. OASIS knock-down does not induce U373 cell apoptosis. (A) U373, A172 and U87 cells were treated with 1 mM TG for the times indicated prior to lysis and immunoblotting for cleaved caspase 3. Rat pancreatic INS-1 insulinoma cells treated with staurosporine were used as a positive control for detection of cleaved caspase 3. (B) U373 cells transfected with 100 nM control (GFP) or OASIS siRNAs for 7 days, then treated or not with 1 mM TG for an additional 48 h. The cells were lysed and immunoblotted for the indicated proteins. Pancreatic INS-1 cells treated with staurosporine (ST, 3 h or 24 h) were used as a positive control for detection of cleaved caspase 3. (C) U87 cells were treated with siRNAs as in (B), then with TG for 16 or 48 h and total protein per well was measured in cell lysates. N = 3 independent experiments. Bars are SEM. doi:10.1371/journal.pone.0054060.gSupporting InformationFigure S1 Expression of selected ER stress and ECM genes inAcknowledgmentsWe thank Children, comments fellowship 1516647 Toronto). Dr. James Rutka and Dr. Christian Smith (Hospital for Sick Toronto) for providing the human glioma cell lines and on the manuscript. RNV was funded by a post-doctoral from the Banting and Best Diabetes Centre (University ofhuman glioblastoma multiforme (GBM) tumors (DOCX)Table S1 Expression of selected ER stress and ECM genes inhuman glioblastoma multiforme (GBM) tumors (DOCX)Author ContributionsConceived and designed the experiments: RV AV. Performed the experiments: RV LZ. Analyzed the data: RV LZ AV. Wrote the paper: RV AV.OASIS in Human Glioma Cells
Dynamins are large GTPases involved in a wide range of cell and organelle fission events. The dynamin superfamily is made up of classical dynamins and dynamin-like proteins. Classical dynamins are critical components of clathrin-mediated endocytosis, where they contribute to the release of newly formed endosomes [1,2,3]. In addition to this well-char.

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