Journal: Cancer Science

Article Title: Thymoquinone inhibits the metastasis of renal cell cancer cells by inducing autophagy via AMPK / mTOR signaling pathway

doi: 10.1111/cas.13808

Figure Lengend Snippet: Thymoquinone (TQ) suppresses migration, invasion and epithelial‐mesenchymal transition (EMT) in renal cell carcinoma (RCC) cells. A, Cell viability in 786‐O and ACHN cells treated with TQ . Cells were cultured with different concentrations (0, 10, 20, 40, 60, 80 and 100 μmol/L) of TQ for 24 or 48 hours. Cell viability was tested using CCK 8 assay. * * P < .01, * ** P < .001 and ### P < .001 vs non‐treated group (0 μmol/L). B, Wound healing assay was conducted in 786‐O and ACHN cells. Cells were cultured with TQ (40 μmol/L); then, the wound healing assay was observed at 0, 12 and 24 hours in 786‐O cells and 0, 24 and 36 hours in ACHN cells. * ** P < .001 vs control group. C, Transwell migration and invasion assay was observed in 786‐O and ACHN cells. Cells were treated with TQ at different concentrations (0, 20, 40 and 60 μmol/L). The migration assay was observed at 24 hours and the invasion assay was observed at 48 hours in both 786‐O and ACHN cells. * P < .05, ** P < .01 and *** P < .001 vs non‐treated group (0 μmol/L). D, The expressions of EMT ‐related proteins in TQ ‐treated RCC cells. Epithelial markers (E‐cadherin) and mesenchymal markers (N‐cadherin, vimentin) were detected by western blot in TQ ‐treated 786‐O and ACHN cells at indicated concentrations and times. All data are presented as mean ± SD of 3 independent experiments

Article Snippet: Human renal carcinoma cell lines ACHN and 786‐O were purchased from ATCC (Manassas, VA, USA).

Techniques: Migration, Cell Culture, CCK-8 Assay, Wound Healing Assay, Control, Invasion Assay, Western Blot

Journal: Cancer Science

Article Title: Thymoquinone inhibits the metastasis of renal cell cancer cells by inducing autophagy via AMPK / mTOR signaling pathway

doi: 10.1111/cas.13808

Figure Lengend Snippet: Thymoquinone (TQ) induces autophagy in renal cell carcinoma (RCC) cells . A, Morphological changes in 786‐O and ACHN cells. RCC cells were treated with 40 μmol/L TQ for 24 hours; then, an inverted microscope was used to observe the morphological changes (×40 and ×100). Arrows point to cytoplasmic vacuoles. B, Autophagosomes and autolysosomes induced by TQ in RCC cells. Cells were transfected with mRFP ‐ EGFP ‐ LC 3 reporter plasmids and treated with 40 μmol/L TQ for 24 hours. The LC 3 puncta was observed by fluorescence microscopy (×200). Yellow arrows point to autophagosomes ( mRFP ‐positive/ EGFP ‐positive) and red arrows point to autolysosomes ( mRFP ‐positive/ EGFP ‐negative). C, Quantification of the number of autophagosomes and autolysosomes per cell. D, The level of LC 3 was analyzed by western blot in 786‐O and ACHN cells after treatment with different concentrations of TQ . All data are presented as mean ± SD of 3 independent experiments

Article Snippet: Human renal carcinoma cell lines ACHN and 786‐O were purchased from ATCC (Manassas, VA, USA).

Techniques: Inverted Microscopy, Transfection, Fluorescence, Microscopy, Western Blot

Journal: Cancer Science

Article Title: Thymoquinone inhibits the metastasis of renal cell cancer cells by inducing autophagy via AMPK / mTOR signaling pathway

doi: 10.1111/cas.13808

Figure Lengend Snippet: Inhibition of autophagy attenuates thymoquinone (TQ)‐induced anti‐tumor effects in renal cell carcinoma (RCC) cells. 786‐O and ACHN cells were mock‐treated or treated with TQ (40 μmol/L) in the presence or absence of 3‐ MA (5 mmol/L); then, the migratory and invasive abilities were detected by transwell assay (A), and the expression of epithelial‐mesenchymal transition (EMT) markers and LC 3 was determined by western blot (B). All data are presented as mean ± SD of 3 independent experiments

Article Snippet: Human renal carcinoma cell lines ACHN and 786‐O were purchased from ATCC (Manassas, VA, USA).

Techniques: Inhibition, Transwell Assay, Expressing, Western Blot

Journal: Cancer Science

Article Title: Thymoquinone inhibits the metastasis of renal cell cancer cells by inducing autophagy via AMPK / mTOR signaling pathway

doi: 10.1111/cas.13808

Figure Lengend Snippet: AMPK / mTOR pathway is involved in anti‐tumor effects of autophagy induction by thymoquinone (TQ) in renal cell carcinoma (RCC) cells. A, 786‐O and ACHN cells were treated with different concentrations (0, 20, 40, 60 μmol/L) of TQ for 24 hours; the levels of t‐ AMPK , p‐ AMPK , t‐ mTOR , p‐ mTOR , p‐S6K and LC 3 were evaluated by western bolt. B, RCC cells were pretreated with AICAR (1 mmol/L) or compound C (50 μmol/L) for 4 hours, and then incubated with TQ (40 μmol/L) for 24 hours. The expression of p‐ AMPK , p‐ mTOR and LC 3 was detected by western bolt. C, D, Then, the migratory and invasive abilities were detected by transwell assay and epithelial‐mesenchymal transition‐related proteins were measured by western blot. All data are presented as mean ± SD of 3 independent experiments

Article Snippet: Human renal carcinoma cell lines ACHN and 786‐O were purchased from ATCC (Manassas, VA, USA).

Techniques: Western Blot, Incubation, Expressing, Transwell Assay

Journal: PeerJ

Article Title: Exosomes in cancer: small vesicular transporters for cancer progression and metastasis, biomarkers in cancer therapeutics

doi: 10.7717/peerj.4763

Figure Lengend Snippet: Overview of the role of exosomes in multiple kinds of cancer.

Article Snippet: ACTB, TUBA1A, FN1, FNLA, CD61, HLA-A, LGALS3BP, Alix, RAB5B, RAB5C, SDCBP, VPF37B, CLTC, ARF1, ANXA2, ANXA5, HSC70, HSP72, RAC1, STOM, MFGE8, MVP, GNA12, PTGFRN, HBA1, tumor susceptibility gene-101 (TSG-101), and Grp94 , The inhoused established human HCC cell lines HKCI-C3 and HKCI-8 cells The hepatocellular carcinoma (HCC) cell line MHCC97L cells The immortalized hepatocyte cell line MIHA cells , qRT-PCR analysis, Ion Torrent Next-Generation Sequencing, and Western blotting , The internalization of exosomes could activate PI3K/AKT and MAPK signaling pathway, and promote secretion of MMP-2 and MMP-9 that favored cell invasion. Also, by proteome analysis Syndecan–syntenin–ALIX is known to support biogenesis of exosomes and the segregation of signaling cargo to these vesicles. The research also detected the components of endosomal protein sorting complex, such as VPS28 and VPS37, whose functions are required for exosome cargo sorting Process. , .

Techniques: Western Blot, Immunofluorescence, AChE Assay, Immunocytochemistry, Inhibition, Chromatin Immunoprecipitation, Marker, Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Biomarker Assay, In Vitro, In Silico, Diagnostic Assay, Isolation, Flow Cytometry, Variant Assay, In Vivo, Sampling, Modification, Transgenic Assay, Migration, Luciferase, Plasmid Preparation, In Vivo Imaging, Purification, Ex Vivo, Activation Assay, Injection, Endothelial Tube Formation Assay, Viability Assay, Matrigel Assay, Immunoprecipitation, Microscopy, Functional Assay, Knock-Out, Mass Spectrometry, Dot Blot, Electron Microscopy, Methylation, Chromatography, Over Expression, Mutagenesis, Microarray, Binding Assay, Transferring, Real-time Polymerase Chain Reaction, Bradford Assay, Transformation Assay, Animal Model, MTT Assay, Fluorescence, FACS, Diffusion-based Assay, Next-Generation Sequencing, Transplantation Assay, Sequencing, Histone Deacetylase Assay, Infection, Incubation, Transmission Assay, Blocking Assay, Luminex, Fractionation, Irradiation, Reporter Assay