Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Upregulated Talin1 synergistically boosts β-estradiol-induced proliferation and pro-angiogenesis of eutopic and ectopic endometrial stromal cells in adenomyosis

doi: 10.1186/s12958-021-00756-7

Figure Lengend Snippet: β-E 2 induced adenomyotic endometrial stromal cell proliferation in vitro. a . Cell proliferation rates of adenomyotic and normal endometrial stromal cells with or without β- E 2 . b . Numbers of formative colonies of adenomyotic and normal endometrial stromal cells with or without β- E 2 . c . and d . CCK-8 assay and colony formation assay were performed to evaluate the effects of β- E 2 on the proliferation of ADS eutopic and ectopic endometrial stromal cells. e . and f . Western blot and gray value analysis of molecular markers for cell division and proliferation including PCNA and Ki67 in ADS endometrial stromal cells treated with E 2 (10nM) or Fulvestrant (a selective estrogen receptor antagonist, 10 nM) or both or neither of them. GAPDH was used as a loading control. Data were presented as mean ± SD. NS no significance, * P < 0.05, ** P < 0.01. ADS_Eu_ESC, adenomyotic eutopic endometrial stromal cells; ADS_Ec_ESC, adenomyotic ectopic endometrial stromal cells

Article Snippet: Subsequently, cells were incubated in fresh medium (as control), β-E 2 (10nM, Sigma, USA), Fulvestrant (10 nM, ICI 182780, a selective ER antagonist, MedChem Express, USA) or β-E 2 plus Fulvestrant for 24 h.

Techniques: In Vitro, CCK-8 Assay, Colony Assay, Western Blot, Control

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Upregulated Talin1 synergistically boosts β-estradiol-induced proliferation and pro-angiogenesis of eutopic and ectopic endometrial stromal cells in adenomyosis

doi: 10.1186/s12958-021-00756-7

Figure Lengend Snippet: β-E 2 induced adenomyotic endometrial stromal cell angiogenesis in vitro. a . b . Assessment of pro-angiogenetic ability of adenomyotic and normal endometrial stromal cells with or without β- E 2 . c . d . Tube and network formation assay of HUVEC cells treated with conditioned media (the whole cell filtrate from ADS_Eu_ESC and ADS_Ec_ESC added with either E 2 (10nM) or Fulvestrant (10 nM) or both. Images were captured when HUVEC were cultured with the conditioned media on Matrigel for 6 h. Branch points of capillaries were counted and analyzed. Scale bar = 100 μm. e . f . Western blot detection of angiogenesis-related proteins including VEGFB and ANGPTL4 in ADS_Eu_ESC and ADS_Ec_ESC cells after being treated with E 2 (10nM) or Fulvestrant (10 nM). GAPDH was used as a loading control. Data were presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001. HUVEC, human umbilical vein endothelial cells

Article Snippet: Subsequently, cells were incubated in fresh medium (as control), β-E 2 (10nM, Sigma, USA), Fulvestrant (10 nM, ICI 182780, a selective ER antagonist, MedChem Express, USA) or β-E 2 plus Fulvestrant for 24 h.

Techniques: In Vitro, Tube Formation Assay, Cell Culture, Western Blot, Control

Journal: Cancers

Article Title: Carcinoma-Associated Fibroblasts Promote Growth of Sox2-Expressing Breast Cancer Cells

doi: 10.3390/cancers12113435

Figure Lengend Snippet: Description of the MCF-7 sublines used in this study.

Article Snippet: LCMF-AnD5 and LF-AnD5 cells were generated side-by-side by exposing AnD5 cells to 20% CAF-CM plus 1 nM fulvestrant (LKT Laboratories, St. Paul, MN, USA) or 1 nM fulvestrant alone, respectively.

Techniques: Generated

Journal: Cancers

Article Title: Carcinoma-Associated Fibroblasts Promote Growth of Sox2-Expressing Breast Cancer Cells

doi: 10.3390/cancers12113435

Figure Lengend Snippet: Short- and long-term exposure of CAF-CM (carcinoma-associated fibroblast-conditioned medium) increases Sox2 expression in breast cancer cells. ( A , B ) MCF-7 parental or MCF-7/AnD3 subline AnD3/M-D1, AnD3/M-D2, and AnD3/M-D3) cells were grown in the presence or absence of CAF-CM for three days, before nuclear protein extracts were examined for Sox2 expression by Western blot analysis. (( C , D ), left panels) Western blot analyses of AnD5, LCM-AnD5, LF-AnD5 and LCMF-AnD5 cells for the proteins as indicated. Before protein extraction, cells were grown in the absence of CAF-CM and fulvestrant for three days. (( C , D ), right panels) Effect of three-day-treatment of LCM-AnD5 and LCMF-AnD5 cells with CAF-CM on the expression of proteins as indicated. ( C , D ) NE = nuclear extract, CE = cytosolic extract, PM = plasma membrane extract, ITGB1 = integrin β1. ( A – D ) Protein stain Fast green (FastG) was used to confirm equal protein loading.

Article Snippet: LCMF-AnD5 and LF-AnD5 cells were generated side-by-side by exposing AnD5 cells to 20% CAF-CM plus 1 nM fulvestrant (LKT Laboratories, St. Paul, MN, USA) or 1 nM fulvestrant alone, respectively.

Techniques: Expressing, Western Blot, Protein Extraction, Clinical Proteomics, Membrane, Staining

Journal: Cancers

Article Title: Carcinoma-Associated Fibroblasts Promote Growth of Sox2-Expressing Breast Cancer Cells

doi: 10.3390/cancers12113435

Figure Lengend Snippet: Knock-down of Sox2 increases apoptotic activity of breast cancer cells and reduces their growth activity. ( A ) Comparison of LF-AnD5 and LCMF-AnD5 growth activities by an ATP-based growth assay. Cells were incubated with 0, 5, 50, or 250 nM fulvestrant for 7 days. ( B , C ) Effect of siSox2, control siRNA (ctrl siRNA) or mock transfection (no siRNA) on growth activity on LCMF-AnD5 and LCM-AnD5 cells. After transfection cells were grown in the presence of CAF-CM, 1 nM fulvestrant or in the absence of either agent (mock) for 4 days. ( A – C ) Each bar represents the average value of five experiments. Statistical analysis was performed with one-way ANOVA, followed by Bonferroni correction for post-hoc analysis. Asterisks indicate statistical significance: ** ( p < 0.01), **** ( p < 0.001), ***** ( p < 0.0001). ( D ) Western blot analysis of cytosolic protein extracts from LCMF-AnD5 and LCM-AnD5 cells for cPARP-1 abundance after transfection with siSox2 or crtl siRNA. FastG (Fast green) staining was used to check for equal protein loading. ( E ) ICC analysis to examine the percentage of cPARP-1-positive cells (left panel). Box plot (right panel) shows the percentage of cPARP-1-positive cells in the presence of siSox2, crtl siRNA, or in the absence of any siRNA. For each condition, 40 fields of view (~200 cells/field) were examined. Statistical analysis was performed by using the Mann-Whitney-U-test. The asterisks indicate statistical significance ( p < 0.0001).

Article Snippet: LCMF-AnD5 and LF-AnD5 cells were generated side-by-side by exposing AnD5 cells to 20% CAF-CM plus 1 nM fulvestrant (LKT Laboratories, St. Paul, MN, USA) or 1 nM fulvestrant alone, respectively.

Techniques: Knockdown, Activity Assay, Comparison, Growth Assay, Incubation, Control, Transfection, Western Blot, Staining, MANN-WHITNEY

Journal: International Journal of Environmental Research and Public Health

Article Title: Protein Kinase CK2 Expression Predicts Relapse Survival in ERα Dependent Breast Cancer, and Modulates ERα Expression in Vitro

doi: 10.3390/ijerph13010036

Figure Lengend Snippet: CK2 inhibition results in increased ERα-dependent ERE-transcriptional activity. T47D cells stably transfected with an ERE-luciferase were incubated in the presence of ( A ) 2.5 or 25 μM TBCA or ( B ) fulvestrant (1 μM) and/or TBCA (25 μM) for 24 h. Luciferase activity was measured was ascertained after 24 h. Each study was performed for at least 3 experimental repeats.

Article Snippet: Stably transfected T47D-ERE-luciferase cells were cultured in phenol red free RPMI 1640 media supplemented with 10% charcoal-dextran treated FBS (Life Technologies, Grand Island, NY USA) for a minimum of 72 h. Subsequently, cells were incubated with TBCA (tetra-bromo-cinnamic acid EMD Millipore, Billerica, MA USA) and/or fulvestrant (Santa Cruz Biotechnologies, Dallas, TX, USA) for 24 h. Cells were then lysed and luciferase activity was assayed using Bright-Glo © (Promega Madison, WI, USA) detection system.

Techniques: Inhibition, Activity Assay, Stable Transfection, Transfection, Luciferase, Incubation