Image_2_Expression Profile and Prognostic Values of HOXA Family Members in Laryngeal Squamous Cell Cancer.TIF

The homeobox A cluster (HOXA) gene family, comprising 11 members, is involved in a wide spectrum of biological functions in human cancers. However, there is little research on the expression profile and prognostic values of HOXA genes in laryngeal squamous cell cancer (LSCC). Based on updated public resources and integrative bioinformatics analysis, we assessed the expression profile and prognostic values of the HOXA family members. Expression and methylation data on HOXA family members were obtained from The Cancer Genome Atlas (TCGA). The prognostic values of HOXA members and clinical features were identified. A gene set enrichment analysis (GSEA) was conducted to explore the mechanism underlying the involvement of HOXA members in LSCC. The associations between tumor immune infiltrating cells (TIICs) and the HOXA family members were evaluated using the Tumor Immune Estimation Resource (TIMER) database. HOXA2 and HOXA4 were downregulated and HOXA7 and HOXA9–13 were upregulated in LSCC. Upregulation of HOXA10, HOXA11, and HOXA13, along with two clinical characteristics (M stage and gender), were associated with a poor LSCC prognosis based on the results of univariate and multivariate Cox proportional hazards regression analyses. Although there were no significant correlations between TIICs and HOXA members, the GSEA results indicated that HOXA members participate in multiple biological processes underlying tumorigenesis. This study comprehensively analyzed the HOXA members, providing insights for further investigation of the HOXA family members as potential targets in LSCC.</p

Image_1_Expression Profile and Prognostic Values of HOXA Family Members in Laryngeal Squamous Cell Cancer.TIF

The homeobox A cluster (HOXA) gene family, comprising 11 members, is involved in a wide spectrum of biological functions in human cancers. However, there is little research on the expression profile and prognostic values of HOXA genes in laryngeal squamous cell cancer (LSCC). Based on updated public resources and integrative bioinformatics analysis, we assessed the expression profile and prognostic values of the HOXA family members. Expression and methylation data on HOXA family members were obtained from The Cancer Genome Atlas (TCGA). The prognostic values of HOXA members and clinical features were identified. A gene set enrichment analysis (GSEA) was conducted to explore the mechanism underlying the involvement of HOXA members in LSCC. The associations between tumor immune infiltrating cells (TIICs) and the HOXA family members were evaluated using the Tumor Immune Estimation Resource (TIMER) database. HOXA2 and HOXA4 were downregulated and HOXA7 and HOXA9–13 were upregulated in LSCC. Upregulation of HOXA10, HOXA11, and HOXA13, along with two clinical characteristics (M stage and gender), were associated with a poor LSCC prognosis based on the results of univariate and multivariate Cox proportional hazards regression analyses. Although there were no significant correlations between TIICs and HOXA members, the GSEA results indicated that HOXA members participate in multiple biological processes underlying tumorigenesis. This study comprehensively analyzed the HOXA members, providing insights for further investigation of the HOXA family members as potential targets in LSCC.</p

Why do public sectors perform high-level green public procurement practice? A new insight with fsQCA approach

Despite extensive studies on drivers for public sector’s green public procurement (GPP) practice, existing studies have not yet introduced configuration analysis based on holistic perspective into this field. This study, based on the Chinese background, focuses on the public sector and develops a conceptual model from a holistic perspective. Survey data are collected and we adopt the fsQCA approach for further testing, aiming to explore which combinations of GPP motivators can induce high-level GPP practice by public sectors. It is found that five configurations can induce high-level GPP practice. According to the core condition, these configurations can be further categorized as two groups: “top management support-dominated” and “non-top management support-dominated”. The holistic perspective contributes to a better understanding for GPP practice and a new explanation for the conflicted conclusions of existing studies. The managerial and policy implications are also helpful to public sector managers and policymakers.</p

Work flow for identifying imatinib response biomarkers through an informatics approach in K562 cells.

<p>Work flow for identifying imatinib response biomarkers through an informatics approach in K562 cells.</p

Additional file 1 of Endometrial cancer (EC) derived G3BP1 overexpression and mutant promote EC tumorigenesis and metastasis via SPOP/ERα axis

Additional file 1: Supplementary Table 1. Primer for qRT-PCR and constructions, siRNA oligonucleotide sequences. Supplementary Fig. 1. Bioinformatics analysis of G3BP1 mRNA and protein. (A) Kaplan-Meier survival curves of EC based on the TCGA cohort. (B) Heat map of clinical characteristics of EC based on TCGA cohort. (C) Heat map of clinical characteristics of EC based on CPTAC cohort. (D) Gene co-expression circle of G3BP1 based on TCGA cohort. (E) KEGG analysis was performed based on CPTAC cohort. (F) Based on CPTAC cohort, R package was used to enrich gene ontology, including BP, molecular MF and CC. Data are shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Supplementary Fig. 2. The promoting effects of G3BP1 and G3BP1 Q392* on the proliferation and metastasis of endometrial carcinoma partly depend on SPOP/ERα axis. (A) AN3-CA and HEC-1-A cells were transfected with indicator plasmids. 48h after transfection, cell lysates were prepared and the protein levels of ERα, SPOP and G3BP1 were determined by western blot. (B) CCK8 cell proliferation analysis was used to detect the proliferation ability of AN3-CA and HEC-1-A cells. (C) Colony formation assay was used to detect the colony formation ability of AN3-CA and HEC-1-A cells. (D) Cell migration assay was used to detect the metastasis ability of AN3-CA and HEC-1-A cells. Data are shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Supplementary Fig. 3. Fulvestrant can reverse the promoting effects of G3BP1 and G3BP1 Q392* on endometrial carcinoma. (A) HEC-1-A cells were transfected with indicator plasmids. The cells were cultured with DMSO and fulvestrant (100nM) in complete medium. 48h after transfection, cell lysates were prepared and the protein levels of ERα, SPOP and G3BP1 were determined by western blot. (B) CCK8 cell proliferation analysis was used to detect the proliferation ability of HEC-1-A cells. (C) Colony formation assay was used to detect the colony formation ability of HEC-1-A cells. (D) Cell migration assay was used to detect the metastasis ability of HEC-1-A cells. Data are shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000

12 predicted imatinib response gene-miRNA pairs based on GEO data.

<p>Functional experiments were conducted on the bolded gene/miR pair.</p><p>12 predicted imatinib response gene-miRNA pairs based on GEO data.</p

Biological consequences of <i>IL8</i> inhibition.

<p>(<b>A</b>) <i>IL8</i> silencing through siRNA and miR-493-5p mimic resulted in significantly slower cellular proliferation as measured by CellTiter-Glo at 48 hours. (*p<0.05, N = 6). Relative cell growth was calculated by normalizing fluorescence indexes of each time point to the 0 hour control. (<b>B</b>) Cellular response to imatinib treatment curves. Cellular response to imatinib was measured using CellTiter-Glo. p<0.0001 by two way ANOVA, N = 6. Percent survival was calculated by normalizing fluorescence indexes of imatinib treated cells to cells treated with vehicle and plotting on a log(2) scale.</p

Relationships among imatinib treatment, and the expression of miR-493-5p and <i>IL8</i> in K562 cells.

<p>(<b>A</b>) Imatinib treatment (1µM for 24 and 48 hours) significantly up-regulated miR-493-5p expression and significantly down-regulated <i>IL8</i> expression in K562 cells (*p<0.05, N = 6). (<b>B</b>) <i>IL8</i> is significantly down-regulated by miR-493-5p mimic at 24 hours after transfection (*p = 0.002, N = 6). Relative fold change was calculated by first normalizing <i>IL8 to B2M</i> and miR-493-5p to <i>RNU6</i> and then comparing 6 and 24 hours data to 0 hour data.</p

VEGF changed the Serine 1179 phosphorylation status in dimers only and did not alter the phosphorylation status of Threonine 497 in eNOS dimers or monomers.

<p>(A) After serum starvation for two hours, BAECs were treated with VEGF (50 ng/ml) for indicated time. The cell lysates were run on SDS-PAGE gels at low temperature. After transferring the protein to nitrocellulose membranes, they were probed with the indicated antibodies. (B) eNOS dimer Ser1179 phosphorylation density was determined by AlphaImager and normalized with control. The results were representative of three independent experiments. (C) The Thr497 phosphorylation density on eNOS dimer and monomer was determined by AlphaImager and normalized with control. The results were represented from three independent experiments (*P<0.05 comparing to control).</p

Inhibition of Hsp90 resulted in eNOS monomerization without change phosphorylation status of Serine 1179 and Threonine 497 in eNOS dimmer.

<p>(A) BAECs were treated with 20 µM geldanamycin for 48 hours and the samples were run SDS-PAGE gel at low temperature and blotted with the indicated antibodies to determine the changes in phosphorylation status of eNOS and dimers and monomers. The eNOS dimer and monomer densities were determined using AlphaImager. And The Ser1179 and Thr497 phosphorylation density on eNOS dimer were also determined using AlphaImager. All results are representative of three independent experiments. * and # P<0.05 comparison with control. (B) BAECs were silenced with scramble siRNA control (scRNA) or Hsp90 (siHsp90) as indicated concentration for 3 days, the cell lysates were blotted with eNOS or Hsp90 antibody respectively. The eNOS monomer and dimer density were measured using AlphaImager and the average of three independent experiments was presented (* and #P<0.05 compared to eNOS dimer and monomer control respectively).</p