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: 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

Journal: Bioengineered

Article Title: Circular RNA circ_0000228 promotes the malignancy of cervical cancer via microRNA-195-5p/ lysyl oxidase-like protein 2 axis.

doi: 10.1080/21655979.2021.1954846

Figure Lengend Snippet: Figure 1. Circ_0000228 was remarkably up-modulated in CC tissues and cell lines. (a) A volcano plot was employed to analyze the differentially expressed circRNAs in the GEO dataset (GSE113696) that were remarkably up- and down-modulated in CC cell lines, compared with normal cervical epithelial cells; (b) A heat maps was utilized to present the significantly highly expressed circRNAs in CC; (c) Circ_0000228 expression in 40 CC tissues/paired adjacent cervical tissues was examined by qRT-PCR; (d) Circ_0000228 expression in 21 patients with stage I CC and 19 patients with stage II CC was analyzed by qRT-PCR, respectively; (e) ZEB1 mRNA expression in 21 patients with stage I CC and 19 patients with stage II CC was analyzed by qRT-PCR, respectively; (f) Circ_0000228 expression in HUCEC and CC cell lines (HeLa, HCC94, SW756, C33A) was detected by qRT-PCR; (g) Circ_0000228 expression in the nucleus and cytoplasm of HCC94 and C33A cells was detected by qRT-PCR. All of the experiments were performed in triplicate. ***P < 0.001.

Article Snippet: Four human CC cell lines (HeLa, HCC94, SW756, and C33A) and human normal cervical epithelial cells (HUCEC) were procured from the American Type Culture Collection (ATCC, Bethesda, MD, USA) and the Chinese Academy of Sciences Committee Type Culture Collection Cell Bank (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: Bioengineered

Article Title: Circular RNA circ_0000228 promotes the malignancy of cervical cancer via microRNA-195-5p/ lysyl oxidase-like protein 2 axis.

doi: 10.1080/21655979.2021.1954846

Figure Lengend Snippet: Figure 3. Circ_0000228 modulated LOXL2 expression through miR-195-5p. (a) A volcano plot was adopted to show the miRNAs that were remarkably up- and down-modulated in CC tissues compared with normal cervical tissue in the GEO dataset (GSE86100); (b) The Venn diagram was used to screen out the differentially expressed miRNAs in GSE86100, which contained complementary binding sites with circ_0000228; (c) Bioinformatics analysis indicated that high LOXL2 expression suggested poor clinical outcome of CC patients. (d) The complementary binding sites of circ_0000228 and miR-195-5p, miR-195-5p and LOXL2 3UTR are presented, and the corresponding luciferase reporter vectors were designed; (E-F) Dual-luciferase reporter gene experiments confirmed that the binding sites between circ_0000228 and miR-195-5p, miR-195-5p and LOXL2 3UTR were functional. (g) MiR-195-5p expression in 40 pairs of CC tissues/adjacent cervical tissues was detected by qRT-PCR; (h) MiR-195-5p expression in stage I and stage II CC patients was detected by qRT-PCR, respectively; (I) MiR-195-5p in HUCEC and CC cell lines (HeLa, HCC94, SW756, C33A) was detected by qRT-PCR.(j) LOXL2 mRNA expression in 40 pairs of CC tissues/adjacent cervical tissues was detected by qRT-PCR; (k) LOXL2 mRNA expression in HUCEC and CC cell lines (HeLa, HCC94, SW756, C33A) was detected by qRT-PCR; (L) LOXL2 mRNA expression in stage 1 and stage 2 CC patients was detected by qRT-PCR; (m-o) Pearson’s correlation analysis of the correlation relationships among circ_0000228 miR-195-5p, and LOXL2 expression in CC tissues; (p) qRT- PCR was employed to detect miR-195-5p expression after co-transfection with circ_0000228 siRNA and miR-195-5pin in CC cell lines; (q) qRT-PCR and Western blot experiments were employed to detect LOXL2 expression after co-transfection with circ_0000228 siRNA and miR-195-5p inhibitors in CC cell lines; All of the experiments were performed in triplicate. *P < 0.05, **P < 0.01 and ***P < 0.001.

Article Snippet: Four human CC cell lines (HeLa, HCC94, SW756, and C33A) and human normal cervical epithelial cells (HUCEC) were procured from the American Type Culture Collection (ATCC, Bethesda, MD, USA) and the Chinese Academy of Sciences Committee Type Culture Collection Cell Bank (Shanghai, China).

Techniques: Expressing, Binding Assay, Luciferase, Functional Assay, Quantitative RT-PCR, Cotransfection, Western Blot

Journal: Cell reports

Article Title: Interferon (IFN)-γ promotes monocyte-mediated lung injury during influenza infection

doi: 10.1016/j.celrep.2022.110456

Figure Lengend Snippet: Key Resources Table

Article Snippet: Primary Bronchial/Tracheal Epithelial Cells; Normal, Human , ATCC , Cat#PCS-300-01.

Techniques: Virus, Recombinant, cDNA Synthesis, SYBR Green Assay, Protein Extraction, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Software, Microscopy, Flow Cytometry, Fluorescence