Journal: Nutrients

Article Title: Essential Oils, Pituranthos chloranthus and Teucrium ramosissimum , Chemosensitize Resistant Human Uterine Sarcoma MES-SA/Dx5 Cells to Doxorubicin by Inducing Apoptosis and Targeting P-Glycoprotein

doi: 10.3390/nu13051719

Figure Lengend Snippet: Effects of Pituranthos chloranthus (PC) and Teucrium ramosissimum Desf. (TR) extracts on normal primary human uterine fibroblast cells (HUF) and primary murine Bone Marrow-Derived Macrophages (BMDM) viability. After treatment of primary HUF and murine BMDM with increasing concentrations (0–100 µg/mL) of PC and TR for 72 h, the percentage of viable cells was assessed using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. ( A ) Dose–response curves of PC-treated HUF (left panel) and TR-treated HUF (right panel). ( B ) Dose–response curves of PC-treated BMDM (left panel) and TR-treated BMDM (right panel). Data are expressed as a mean percentage of control growth ± Standard Deviation (SD) of two representative experiments ( n = 6 replicates per concentration).

Article Snippet: Primary human uterine fibroblast normal cells (HUF) were obtained from the ATCC.

Techniques: Derivative Assay, MTT Assay, Control, Standard Deviation, Concentration Assay

Journal: Allergy, asthma & immunology research

Article Title: Serum Amyloid A1: A Biomarker for Neutrophilic Airway Inflammation in Adult Asthmatic Patients.

doi: 10.4168/aair.2022.14.1.40

Figure Lengend Snippet: Fig. 1. The production of SAA1 from AECs and the effects of SAA1 on proinflammatory cytokine production. SAA1 proteins were produced from AECs (SAECs and A549, respectively) in response to various stimuli (A, B), and in the coculture with PBNs, PBEs (C, D). SAA1-induced release of IL-6, IL-8 and S100A9 from AECs (SAEC and A549, respectively) (E-K). Data are presented as means ± standard error of mean (n = 6). P values were calculated by using one-way analysis of variance with Bonferroni's post hoc test. SAA1 enhanced the expression of p-ERK and p-p38 MAPK (L) in AECs (SAECs and A549, respectively). IL, interleukin; PBEs, peripheral blood eosinophils; PBNs, peripheral blood neutrophils; dex, dexamethasone; S100A9, S100 calcium-binding protein A9; p-ERK, phospho-extracellular signal-regulated kinase; ERK, extracellular signal-regulated kinase; p-p38 MAPK, phospho-p38 mitogen-activated protein kinase; MAPK, mitogen-activated protein kinase; AECs, airway epithelial cells; SAEC, small airway epithelial cells. *P < 0.050, †P < 0.010, ‡P < 0.001, ns = not significant.

Article Snippet: Epithelial cell culture and stimulation Two kinds of AECs were used in this study, primary small airway epithelial cells (SAECs) and human AEC line (A549), both of which were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Produced, Expressing, Binding Assay

Journal: Nucleic acids research

Article Title: Sequestration of ribosomal subunits as inactive 80S by targeting eIF6 limits mitotic exit and cancer progression.

doi: 10.1093/nar/gkae1272

Figure Lengend Snippet: Figure 9. Ov ere xpression of eIF6 observ ed primarily in high-grade in v asiv e bladder and breast cancers. ( A and B ) Western blot probed for eIF6 in human bladder cancer cell lines and healthy (normal) bladder epithelial cells. β-Tubulin used as loading control. Blots shown in panel (A) were quantitated and eIF6 le v els w ere normaliz ed to loading control and plotted (panel B) as FC o v er eIF6 le v els in normal bladder epithelial cells. Values indicate standard error of the mean from four independent experiments and significant differences for T24 ( P = 0.0065), UMUC3 ( P = 0.0132) and HT1197 ( P = 0.0017) determined by an unpaired t wo-t ailed t -test. ( C ) Images represent eIF6 expression in patient-derived benign and high-grade tumors by immunohistochemistry using anti-eIF6 antibody. Enlarged inset shows the presence of eIF6 in nucleoli and cytoplasm in high-grade cancers. ( D ) Images shown in panel (C) were quantitated and eIF6 expression in patient-matched benign tissues relative to high-grade cancers (six patients) were plotted. Significant differences were determined using an unpaired t wo-t ailed t -test ( P = 0.0063). ( E ) Plot shows eIF6 expression in unmatched benign tissues, high-grade and low-grade bladder cancers derived from patients. ( F and G ) Western blot represents high levels of eIF6 in high-grade invasive human triple negative breast cancer cell line. β-Tubulin used as loading control. Blots shown in panel (F) were quantitated and eIF6 levels were normalized to loading control and plotted (G) as FC o v er eIF6 le v els in normal (healthy) HME-1 cells. Values indicate standard error of the mean from four independent experiments and significant differences for MDA-MB-231 ( P = 0.001) determined by an unpaired t wo-t ailed t -test.

Article Snippet: The human bladder cancer cell lines HT1376, HT1197, UM-UC-3 and TCCSUP were maintained in EMEM media with 10% FBS and Penn / Strep, T-24 cell lines were maintained in McCoy’s 5A medium with 10% FBS and Penn / Strep and primary bladder epithelial (A / T / N): normal human (BdEC) cells were maintained in bladder epithelial cell basal medium (ATCC) supplemented with bladder epithelial g c i g b D W C [ a p 1 fl d t ( m a p c w m ( b l o β S 0 p B S C C e d s i S B m B s a ( S B a p a m t w s i i H 7 w f v D ow nloaded from https://academ ic.oup.com /nar/advance-article/doi/10.1093/nar/gkae1272/7933602 by guest on 30 D ecem ber 2024 rowth factors.

Techniques: Western Blot, Control, Expressing, Derivative Assay, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: High-Content Imaging and Machine Learning Classify Phenotypical Change in Coronary Artery Endothelial Cells Caused by BPS

doi: 10.3390/ijms27073259

Figure Lengend Snippet: Representative high-content microscopy images of human coronary artery endothelial cells (HCAEC) exposed to vehicle control (CTRL) or 0.1 µM Bisphenol S (BPS) for 96 h and stained using the PhenoVue Cell Painting assay. For each condition, a representative field acquired at 40× magnification and a higher-magnification inset are shown. Rows correspond to the individual fluorescence channels: Hoechst 33342 (nuclei), PhenoVue Fluor 488 Concanavalin A (endoplasmic reticulum and intracellular membranes), PhenoVue 512 nucleic acid stain (RNA/nucleoli), PhenoVue Fluor 555 wheat germ agglutinin (plasma membrane), PhenoVue 641 mitochondrial stain (mitochondria), and the merged image. White boxes represent the part of the image used for the related inset. Scale bar: 50 µm, 40× objective.

Article Snippet: Primary human coronary artery endothelial cells (HCAEC; ATCC ® PCS-100-020TM, Innovation, VA, USA) were cultured according to the supplier’s recommendations.

Techniques: Microscopy, Control, Staining, Fluorescence, Clinical Proteomics, Membrane

Journal: Molecular medicine reports

Article Title: Long non‑coding RNA SNHG14 affects the proliferation and apoptosis of childhood acute myeloid leukaemia cells by modulating the miR‑193b‑3p/MCL1 axis.

doi: 10.3892/mmr.2020.11729

Figure Lengend Snippet: Figure 1. SNHG14 gene expression is upregulated in bone marrow tissues of patients with AML and AML cell lines. (A) Relative expression of SNHG14 in 57 AML bone marrow tissues and NBM. (B) Relative expression of SNHG14 in AML cell lines and human normal bone marrow CD34+ cells. **P<0.01 vs. CD34+ cells. The 2‑ΔΔCq method was used to analyse the relative mRNA expression level using GAPDH as an internal control. AML, acute myeloid leukaemia; NBM, normal marrow tissues; SNHG14, small nucleolar RNA host gene 14.

Article Snippet: Human normal bone marrow CD34+ cells and AML cell lines (MV‐4‐11, AML‐193, HL‐60, and KG‐1 cells) were obtained from the American Type Culture Collection and cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C.

Techniques: Gene Expression, Expressing, Control

Journal: Molecular medicine reports

Article Title: Long non‑coding RNA SNHG14 affects the proliferation and apoptosis of childhood acute myeloid leukaemia cells by modulating the miR‑193b‑3p/MCL1 axis.

doi: 10.3892/mmr.2020.11729

Figure Lengend Snippet: Figure 3. miR‑193b‑3p is a target of SNHG14 in AML cells. (A) Starbase was used to predict the binding site between SNHG14 and miR‑193b‑3p. (B) Relative expression of miR‑193b‑3p in MV‑4‑11 and AML‑193 cells following SNHG14 silencing. **P<0.01 vs. blank control. (C) The target association between SNHG14 and miR‑193b‑3p was determined using an RNA immunoprecipitation assay. **P<0.01 vs. Anti‑IgG. (D) The target association between SNHG14 and miR‑193b‑3p was determined using a dual luciferase reporter gene assay. **P<0.01 vs. miR‑NC. (E) Relative expression of miR‑193b‑3p in 57 AML bone marrow tissues and NBM was detected by RT‑qPCR. (F) Spearman's correlation analysis was performed to evaluate the correlation between SNHG14 and miR‑193b‑3p expression. (G) Relative expression of miR‑193b‑3p in AML cell lines and human normal bone marrow CD34+ cells. **P<0.01 vs. CD34+ cells. The 2‑ΔΔCq method was used to analyse the relative mRNA expression level using U6 as an internal control. SNHG14, small nucleolar RNA host gene 14; miR, microRNA; si, small interfering RNA; NC, negative control; wt, wildtype; mut, mutated; NBM, normal marrow tissues; AGO2, protein argonaute‑2; AML, acute myeloid leukaemia.

Article Snippet: Human normal bone marrow CD34+ cells and AML cell lines (MV‐4‐11, AML‐193, HL‐60, and KG‐1 cells) were obtained from the American Type Culture Collection and cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C.

Techniques: Binding Assay, Expressing, Control, RNA Immunoprecipitation, Luciferase, Reporter Gene Assay, Small Interfering RNA, Negative Control

Journal: Molecular medicine reports

Article Title: Long non‑coding RNA SNHG14 affects the proliferation and apoptosis of childhood acute myeloid leukaemia cells by modulating the miR‑193b‑3p/MCL1 axis.

doi: 10.3892/mmr.2020.11729

Figure Lengend Snippet: Figure 5. miR‑193b‑3p targets MCL1 in AML cells. (A) TargetScan was used to predict the binding site between miR‑193b‑3p and MCL1. (B) A dual luciferase reporter gene assay was employed to verify the target association between miR‑193b‑3p and MCL1. **P<0.01 vs. miR‑NC. (C) Relative expression of MCL1 in 57 AML bone marrow tissues and NBM was detected by RT‑qPCR. (D) Spearman's correlation analysis was performed to evaluate the correlation between MCL1 and miR‑193b‑3p expression. (E) Spearman's correlation analysis was performed to evaluate the correlation between MCL1 and SNHG14 expression. (F) Relative expression of miR‑193b‑3p in AML cell lines and human normal bone marrow CD34+ cells. **P<0.01 vs. CD34+ cells. The 2‑ΔΔCq method was used to analyse the relative mRNA expression level using GAPDH as an internal control. SNHG14, small nucleolar RNA host gene 14; miR, microRNA; NC, negative control; wt, wildtype; mut, mutated; NBM, normal marrow tissues; AML, acute myeloid leukaemia; MCL1, MCL1 apoptosis regulator BCL2 family member.

Article Snippet: Human normal bone marrow CD34+ cells and AML cell lines (MV‐4‐11, AML‐193, HL‐60, and KG‐1 cells) were obtained from the American Type Culture Collection and cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C.

Techniques: Binding Assay, Luciferase, Reporter Gene Assay, Expressing, Control, Negative Control

Journal: Frontiers in pharmacology

Article Title: The Anticancer Effects of the Pro-Apoptotic Benzofuran-Isatin Conjugate (5a) Are Associated With p53 Upregulation and Enhancement of Conventional Chemotherapeutic Drug Efficiency in Colorectal Cancer Cell Lines.

doi: 10.3389/fphar.2022.923398

Figure Lengend Snippet: FIGURE 7 | Compound 5a decreases HT29 and SW620 cell viability without affecting the viability of normal colon epithelial CCD 841 cells and enhances irinotecan (IRI), 5-fluorouracil (5-FU), and oxaliplatin (OXA) cytotoxic effects on HT29 and SW620 cells. (A) HT29, SW620, and the normal colon epithelial CCD 841 cell lines were exposed to different concentrations (5-10-20 μM) of Compound 5a for 24 h. Cell viability was measured by the MTT assay at 540 nm regarding the cellular metabolic activity. Bar graph showing the cell viability percentage and the data are expressed as mean ± SD (n = 3). ***p < 0.001 and ****p < 0.0001 vs. Control. Half-maximal inhibitory concentrations (IC50) of Compound 5a on HT29 and SW620 cell viability were determined. HT29 (B,C) and SW620 (D,E) cells were treated with different concentrations of the chemotherapeutic drugs IRI, 5-FU, and OXA for 24 h in the presence (C,E) or absence (B,D) of various concentrations (5-10-20 μM) of Compound 5a. Cell cytotoxicity was measured by the MTT assay at 540 nm. Bar graph showing the cell viability percentage and the data are expressed as mean ± SD (n = 3). Half- maximal inhibitory concentrations (IC50) of each chemotherapeutic drug on HT29 and SW620 cell viability were also determined.

Article Snippet: Human normal colon epithelial cell (CCD841 CoTr), colorectal adenocarcinoma HT29, and mCRC SW620 cell lines were obtained from American Type Culture Collection (ATCC, Manassas, VA, United States) and grown in a complete medium composed of DMEM supplemented with 10% heatinactivated fetal bovine serum (FBS), 100 μg/ml streptomycin, 100 IU/ml penicillin and 2 mmol/l L-glutamine.

Techniques: MTT Assay, Activity Assay, Control