Journal: Endocrinology

Article Title: TXNIP Links Anticipatory Unfolded Protein Response to Estrogen Reprogramming Glucose Metabolism in Breast Cancer Cells.

doi: 10.1210/endocr/bqab212

Figure Lengend Snippet: Figure 5. IRE1 activity is required for estrogen suppression of TXNIP expression. A, T47D cells were hormone-stripped for 2 days and then treated with or without 10nM E2 or 10nM E2 plus 100nM ICI182,780. After 48 hours, the cells were harvested for Western blot analysis to examine the effect of estrogen on UPR (right panel). Left panel is the diagram of 3 branches of UPR. B, C, T47D cells were hormone-stripped for 2 days and then treated with or without 10nM E2 alone or plus 1µM GSK2656157, 20µM 4µ8C, or 1µM KIRA6 for 31 hours, respectively. The cells were harvested for Western blot analysis. (D) HCC1428 cells were hormone-stripped for 2 days and then treated with 10nM E2 alone or plus 1µM GSK2656157 or 20µM 4µ8C for 31 hours. Cells were harvested for Western blot analysis. E, T47D cells were cultured in hormone-depleted medium and transfected with siRNA nega- tive control, PERK siRNA, IRE1 siRNA, or ATF6 siRNA. At 24 hours posttransfection, the cells were treated with DMSO or 10nM E2. After 48 hours E2 treatment, the cells were harvested for Western blot analysis.

Article Snippet: TXNIP siRNA (sc-44943), ERα siRNA (sc-29305), PERK siRNA (sc-36213), IRE1 siRNA (sc-40705), ATF6 siRNA (sc37699) and XBP1 siRNA (sc-38627) were purchased from Santa Cruz.

Techniques: Activity Assay, Expressing, Western Blot, Cell Culture, Transfection, Control

Journal: Endocrinology

Article Title: TXNIP Links Anticipatory Unfolded Protein Response to Estrogen Reprogramming Glucose Metabolism in Breast Cancer Cells.

doi: 10.1210/endocr/bqab212

Figure Lengend Snippet: Figure 6. IRE1 activity is required for estrogen-induced glucose uptake and cell proliferation. A, T47D cells were hormone-stripped for 3 days and then treated with or without 10nM E2 alone or plus 1µM GSK2656157, 20µM 4µ8C, or 1µM KIRA6 for 30 hours. The cultured medium was collected for metabolite profile analysis. The glucose uptake and lactate production were normalized to the protein concentration. a, P < 0.05 vs DMSO; b, P < 0.05 vs E2; n.s, P > 0.05 vs DMSO. B, C, T47D cells were cultured in hormone-depleted medium and transfected with siRNA negative control, PERK siRNA, IRE1 siRNA, or ATF6 siRNA. At 24 hours posttransfection, the cells were treated with DMSO or 10nM E2. After 48 hours, the cells were harvested for Western blot analysis (B), and the cultured medium was collected for metabolite profile analysis (C). The glucose uptake and lactate production were normalized to protein concentration. a, P < 0.05 vs siNC of DMSO group; b, P < 0.05 vs siPERK of DMSO group; c, P < 0.05 vs siIRE1 of DMSO group; d, P < 0.05 vs siATF6 of DMSO group; n.s, P > 0.05 vs siIRE1 of DMSO group. D, E, Effect of GSK2656157 and 4µ8C on cell viability of T47D cells in the absence or presence of E2. T47D cells were seeded into 24-well plates after being hormone-stripped for 2 days and continued to culture in hormone- depleted medium. After overnight, the cells were treated with DMSO or 10nM E2. Cell proliferation was measured by the MTT assay at the indicated time after E2 treatment (D). At day 6, the cells were fixed and stained with crystal violet (E). Images were taken using a scanner.

Article Snippet: TXNIP siRNA (sc-44943), ERα siRNA (sc-29305), PERK siRNA (sc-36213), IRE1 siRNA (sc-40705), ATF6 siRNA (sc37699) and XBP1 siRNA (sc-38627) were purchased from Santa Cruz.

Techniques: Activity Assay, Cell Culture, Protein Concentration, Transfection, Negative Control, Western Blot, MTT Assay, Staining

Journal: Oncology Letters

Article Title: miR-20a-5p inhibits endometrial cancer progression by targeting janus kinase 1

doi: 10.3892/ol.2021.12688

Figure Lengend Snippet: Expression of Jak1 in human EC tissues. (A) Jak1 was upregulated in human EC tissues. (B) Jak1 mRNA expression was negatively correlated with miR-20a-5p expression in human EC tissues. *P<0.05. miR, microRNA; EC, endometrial cancer; Jak, janus kinase 1.

Article Snippet: After washing in Tris-buffered saline-Tween-20 (0.05%) solution, membranes were incubated with mouse monoclonal primary antibodies against Jak1 (1:400; cat. no. sc376996; Santa Cruz Biotechnology, Inc.) and GAPDH (1:1,000; cat. no. sc365062; Santa Cruz Biotechnology, Inc.) at 4°C overnight.

Techniques: Expressing

Journal: Oncology Letters

Article Title: miR-20a-5p inhibits endometrial cancer progression by targeting janus kinase 1

doi: 10.3892/ol.2021.12688

Figure Lengend Snippet: miR-20a-5p directly targeted Jak1. (A) Luciferase activity of wt-Jak1 3′UTR was significantly decreased in cells transfected with miR-20a-5p-mimic. (B) Effect of miR-20a-5p on Jak1 mRNA and protein expression. *P<0.05 and # P<0.01. miR, microRNA; EC, endometrial cancer; Jak, janus kinase 1; wt, wild-type; mut, mutant; NC, negative control.

Article Snippet: After washing in Tris-buffered saline-Tween-20 (0.05%) solution, membranes were incubated with mouse monoclonal primary antibodies against Jak1 (1:400; cat. no. sc376996; Santa Cruz Biotechnology, Inc.) and GAPDH (1:1,000; cat. no. sc365062; Santa Cruz Biotechnology, Inc.) at 4°C overnight.

Techniques: Luciferase, Activity Assay, Transfection, Expressing, Mutagenesis, Negative Control

Journal: Oncology Letters

Article Title: miR-20a-5p inhibits endometrial cancer progression by targeting janus kinase 1

doi: 10.3892/ol.2021.12688

Figure Lengend Snippet: Jak1 reversed the effects of miR-20a-5p on cell viability, cell invasion and adhesion. Expression of (A) Jak1 protein and (B) miR-20a-5p in Ishikawa cells following transfection with miR-20a-5p-mimic and Jak1-pcDNA3.1. Jak1 overexpression abolished the effects of miR-20a-5p-mimic on (C) cell proliferation, (D) cell invasive ability (magnification, ×400) and (E) cell adhesion. *P<0.05 and # P<0.01 vs. NC + pc DNA3.1 group; & P<0.05 and @ P<0.01 vs. miR-20a-5p + pc DNA3.1 group. miR, microRNA; EC, endometrial cancer; Jak, janus kinase 1; wt, wild-type; mut, mutant; NC, negative control.

Article Snippet: After washing in Tris-buffered saline-Tween-20 (0.05%) solution, membranes were incubated with mouse monoclonal primary antibodies against Jak1 (1:400; cat. no. sc376996; Santa Cruz Biotechnology, Inc.) and GAPDH (1:1,000; cat. no. sc365062; Santa Cruz Biotechnology, Inc.) at 4°C overnight.

Techniques: Expressing, Transfection, Over Expression, Mutagenesis, Negative Control