Why do supply chain technologies sometimes fail to improve a firm’s performance?

Successful implementation depends on the reason why the technology was adopted in the first place, write Zhongzhi Liu, Daniel Prajogo, and Adegoke Ok

Additional file 1: of ERK is a negative feedback regulator for IFN-γ/STAT1 signaling by promoting STAT1 ubiquitination

Figure S1. The expression of STAT1 in ESCC cell lines has no change via 5-Aza treatments. A. EC1, KYSE150 cell lines were treated with 10 μM of 5-Aza for 24 h, RNA level of STAT1 was analyzed by qRT-PCR. B. EC1, KYSE150 cell lines were treated with 10 μM of 5-Aza for 24 h, Western blot analysis of p-STAT1 and STAT1 in total cell lysates were shown. (JPG 695 kb

Image_4_Functional Plasticity of Gamma Delta T Cells and Breast Tumor Targets in Hypoxia.tiff

<p>Interactions between immune and tumor cells in the tumor microenvironment (TME) often impact patient outcome, yet remain poorly understood. In addition, the effects of biophysical features such as hypoxia [low oxygen (O₂)] on cells within the TME may lead to tumor evasion. Gamma delta T cells (γδTcs) naturally kill transformed cells and are therefore under development as immunotherapy for various cancers. Clinical trials have proven the safety of γδTc immunotherapy and increased circulating γδTc levels correlate with improved patient outcome. Yet, the function of γδTc tumor infiltrating lymphocytes in human breast cancer remains controversial. Breast tumors can be highly hypoxic, thus therapy must be effective under low O₂ conditions. We have found increased infiltration of γδTc in areas of hypoxia in a small cohort of breast tumors; considering their inherent plasticity, it is important to understand how hypoxia influences γδTc function. In vitro, the cell density of expanded primary healthy donor blood-derived human γδTc decreased in response to hypoxia (2% O₂) compared to normoxia (20% O₂). However, the secretion of macrophage inflammatory protein 1α (MIP1α)/MIP1β, regulated on activation, normal T cell expressed and secreted (RANTES), and CD40L by γδTc were increased after 40 h in hypoxia compared to normoxia concomitant with the stabilization of hypoxia inducible factor 1-alpha protein. Mechanistically, we determined that natural killer group 2, member D (NKG2D) on γδTc and the NKG2D ligand MHC class I polypeptide-related sequence A (MICA)/B on MCF-7 and T47D breast cancer cell lines are important for γδTc cytotoxicity, but that MIP1α, RANTES, and CD40L do not play a direct role in cytotoxicity. Hypoxia appeared to enhance the cytotoxicity of γδTc such that exposure for 48 h increased cytotoxicity of γδTc against breast cancer cells that were maintained in normoxia; conversely, breast cancer lines incubated in hypoxia for 48 h prior to the assay were largely resistant to γδTc cytotoxicity. MICA/B surface expression on both MCF-7 and T47D remained unchanged upon exposure to hypoxia; however, ELISAs revealed increased MICA shedding by MCF-7 under hypoxia, potentially explaining resistance to γδTc cytotoxicity. Despite enhanced γδTc cytotoxicity upon pre-incubation in hypoxia, these cells were unable to overcome hypoxia-induced resistance of MCF-7. Thus, such resistance mechanisms employed by breast cancer targets must be overcome to develop more effective γδTc immunotherapies.</p

Correlation between Bcl-2, Bcl-xL, survivin, cyclin D1, p21 and STAT1 protein expression in 62 ESCC samples.

<p>P value for Pearson's X² test; * p<0.05</p><p>Correlation between Bcl-2, Bcl-xL, survivin, cyclin D1, p21 and STAT1 protein expression in 62 ESCC samples.</p

β-catenin decreased USP2a protein expression in MCL cells.

<p>A and B) two MCL cell lines were treated with two different siRNA β-catenin species or scramble siRNA, and the expression of phospho-mTor (ser2481) and USP2a were evaluated by Western blots. siRNA treatment induced a substantial decrease in the levels of the USP2a protein in both cell lines. In contrast, no appreciable modulation of phospho-mTor (as a surrogate marker of mTor activation) was observed. Triplicate experiments were performed and represented results are illustrated.</p

Image_2_Functional Plasticity of Gamma Delta T Cells and Breast Tumor Targets in Hypoxia.tiff

<p>Interactions between immune and tumor cells in the tumor microenvironment (TME) often impact patient outcome, yet remain poorly understood. In addition, the effects of biophysical features such as hypoxia [low oxygen (O₂)] on cells within the TME may lead to tumor evasion. Gamma delta T cells (γδTcs) naturally kill transformed cells and are therefore under development as immunotherapy for various cancers. Clinical trials have proven the safety of γδTc immunotherapy and increased circulating γδTc levels correlate with improved patient outcome. Yet, the function of γδTc tumor infiltrating lymphocytes in human breast cancer remains controversial. Breast tumors can be highly hypoxic, thus therapy must be effective under low O₂ conditions. We have found increased infiltration of γδTc in areas of hypoxia in a small cohort of breast tumors; considering their inherent plasticity, it is important to understand how hypoxia influences γδTc function. In vitro, the cell density of expanded primary healthy donor blood-derived human γδTc decreased in response to hypoxia (2% O₂) compared to normoxia (20% O₂). However, the secretion of macrophage inflammatory protein 1α (MIP1α)/MIP1β, regulated on activation, normal T cell expressed and secreted (RANTES), and CD40L by γδTc were increased after 40 h in hypoxia compared to normoxia concomitant with the stabilization of hypoxia inducible factor 1-alpha protein. Mechanistically, we determined that natural killer group 2, member D (NKG2D) on γδTc and the NKG2D ligand MHC class I polypeptide-related sequence A (MICA)/B on MCF-7 and T47D breast cancer cell lines are important for γδTc cytotoxicity, but that MIP1α, RANTES, and CD40L do not play a direct role in cytotoxicity. Hypoxia appeared to enhance the cytotoxicity of γδTc such that exposure for 48 h increased cytotoxicity of γδTc against breast cancer cells that were maintained in normoxia; conversely, breast cancer lines incubated in hypoxia for 48 h prior to the assay were largely resistant to γδTc cytotoxicity. MICA/B surface expression on both MCF-7 and T47D remained unchanged upon exposure to hypoxia; however, ELISAs revealed increased MICA shedding by MCF-7 under hypoxia, potentially explaining resistance to γδTc cytotoxicity. Despite enhanced γδTc cytotoxicity upon pre-incubation in hypoxia, these cells were unable to overcome hypoxia-induced resistance of MCF-7. Thus, such resistance mechanisms employed by breast cancer targets must be overcome to develop more effective γδTc immunotherapies.</p

Image_1_Functional Plasticity of Gamma Delta T Cells and Breast Tumor Targets in Hypoxia.tif

<p>Interactions between immune and tumor cells in the tumor microenvironment (TME) often impact patient outcome, yet remain poorly understood. In addition, the effects of biophysical features such as hypoxia [low oxygen (O₂)] on cells within the TME may lead to tumor evasion. Gamma delta T cells (γδTcs) naturally kill transformed cells and are therefore under development as immunotherapy for various cancers. Clinical trials have proven the safety of γδTc immunotherapy and increased circulating γδTc levels correlate with improved patient outcome. Yet, the function of γδTc tumor infiltrating lymphocytes in human breast cancer remains controversial. Breast tumors can be highly hypoxic, thus therapy must be effective under low O₂ conditions. We have found increased infiltration of γδTc in areas of hypoxia in a small cohort of breast tumors; considering their inherent plasticity, it is important to understand how hypoxia influences γδTc function. In vitro, the cell density of expanded primary healthy donor blood-derived human γδTc decreased in response to hypoxia (2% O₂) compared to normoxia (20% O₂). However, the secretion of macrophage inflammatory protein 1α (MIP1α)/MIP1β, regulated on activation, normal T cell expressed and secreted (RANTES), and CD40L by γδTc were increased after 40 h in hypoxia compared to normoxia concomitant with the stabilization of hypoxia inducible factor 1-alpha protein. Mechanistically, we determined that natural killer group 2, member D (NKG2D) on γδTc and the NKG2D ligand MHC class I polypeptide-related sequence A (MICA)/B on MCF-7 and T47D breast cancer cell lines are important for γδTc cytotoxicity, but that MIP1α, RANTES, and CD40L do not play a direct role in cytotoxicity. Hypoxia appeared to enhance the cytotoxicity of γδTc such that exposure for 48 h increased cytotoxicity of γδTc against breast cancer cells that were maintained in normoxia; conversely, breast cancer lines incubated in hypoxia for 48 h prior to the assay were largely resistant to γδTc cytotoxicity. MICA/B surface expression on both MCF-7 and T47D remained unchanged upon exposure to hypoxia; however, ELISAs revealed increased MICA shedding by MCF-7 under hypoxia, potentially explaining resistance to γδTc cytotoxicity. Despite enhanced γδTc cytotoxicity upon pre-incubation in hypoxia, these cells were unable to overcome hypoxia-induced resistance of MCF-7. Thus, such resistance mechanisms employed by breast cancer targets must be overcome to develop more effective γδTc immunotherapies.</p

Kaplan-Meier curves of esophageal cancer patients in different positive and negative sub-groups.

<p>(a) Bcl-2-positive patients have longer survival than negative groups. (b), (c), and (e) No significant correlation between overall survival and the expression level of Bcl-xL, survivin nor cyclinD1. (d) Significant correlation between overall survival and the expression level of p21 protein levels when the two groups were defined as p21-positive or p21-negative.</p

FASN inhibition decreased β-catenin expression.

<p>The MCL cell line Mino was treated with Orlistat (A) or siRNA (B) to inhibit or knock-down FASN, respectively. The β-catenin protein expression was determined by western blot after 48 hours of treatment. FASN inhibition by either Orlistat or siRNA substantially decreased the protein expression of β-catenin. Triplicate experiments were performed.</p

Gene transfection of <i>STAT1C</i> upregulated apoptosis in ESCC cell lines.

<p>Using Western blot analysis, the gene transfection of <i>STAT1C</i> in SHEE, NE3, KYSE150 and KYSE510 cells was shown to be effective, since the levels of STAT1, phospho-STAT1 and FLAG were dramatically increased 2 days after STAT1C transfection. By western blots, gene transfection of STAT1C into these cell lines down-regulated several pro-apoptotic proteins (including BCL-2, BCL-xL, survivin), and promoted G1 cell-cycle arrest by decreasing cyclin D1 and increasing p21waf1. Cell lysates were collected 2 days after the gene transfection of STAT1C in all the cell lines.</p