Journal: Oncology Reports

Article Title: WYE-354 restores Adriamycin sensitivity in multidrug-resistant acute myeloid leukemia cell lines

doi: 10.3892/or.2019.7093

Figure Lengend Snippet: Dose-response curves of K562, K562/Adr200 and K562/Adr500 cells to drug treatment. Dose-response curves of cells treated with increasing concentrations of (A) Adr, (B) daunorubicin, (C) idarubicin and (D) etoposide for 48 h demonstrating different fold resistance. (E) Adr florescence in K562/Adr200 and K562/Adr500 cells was decreased compared with that in K562 cells following exposure to 10 µM Adr for 2 h. (F) Reverse transcription-quantitative polymerase chain reaction analysis revealed increased mRNA levels of ABCB1 in K562/Adr200 and K562/Adr500 cells. (G) Western blot showing increased protein expression of ABCB1 in K562/Adr200 and K562/Adr500 cells. **P<0.005 compared with K562. Adr, adriamycin; ABCB1, ATP-binding cassette subfamily B member 1.

Article Snippet: The Adriamycin (Adr)-resistant cell lines K562/Adr200 and K562/Adr500 were generated by culturing K562 cells (CLS Cell Lines Service GmbH, Eppelheim, Germany) in step-wise incremental doses of Adr, ranging between 0.002 and 0.5 μM over a period of 2 months at 37°C with 5% CO 2 in a humidified incubator.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Binding Assay

Journal: Oncology Reports

Article Title: WYE-354 restores Adriamycin sensitivity in multidrug-resistant acute myeloid leukemia cell lines

doi: 10.3892/or.2019.7093

Figure Lengend Snippet: Effects of WYE-354 on Adr cytotoxicity in resistant cells. (A) Structure of WYE-354. (B) WYE-354 IC 50 in the parent and resistant cell lines. Dose-response curves of (C) K562, (D) K562/Adr200 and (E) K562/Adr500 cells following treatment with 0.01–100 µM Adr for 48 h in the presence of 0.2 and 1 µM WYE-354, showing a decrease in Adr IC 50 in the Adr-resistant cell lines. Adr, adriamycin; IC 50 , concentration producing 50% decrease in cell viability.

Article Snippet: The Adriamycin (Adr)-resistant cell lines K562/Adr200 and K562/Adr500 were generated by culturing K562 cells (CLS Cell Lines Service GmbH, Eppelheim, Germany) in step-wise incremental doses of Adr, ranging between 0.002 and 0.5 μM over a period of 2 months at 37°C with 5% CO 2 in a humidified incubator.

Techniques: Concentration Assay

Journal: Oncology Reports

Article Title: WYE-354 restores Adriamycin sensitivity in multidrug-resistant acute myeloid leukemia cell lines

doi: 10.3892/or.2019.7093

Figure Lengend Snippet: ABCB1-ATPase activity and protein expression of ABCB1. (A) Change in RLU of ABCB1 membranes when incubated in the presence of increasing doses of WYE-354 (0–50 µM) and 500 µM verapamil in triplicate for 60 min at 37°C. (B) ATPase activity of ABCB1 as the quantity of ATP consumed demonstrated the stimulatory effect of WYE-354. (C) Western blot images showing difference in protein expression in K562/Adr500 cells when treated with 1 µM WYE-354 for 12, 24 and 48 h, and when treated with 5 and 10 µM of WYE-354 for 24 h. ABCB1, ABCB1, ATP-binding cassette subfamily B member 1.

Article Snippet: The Adriamycin (Adr)-resistant cell lines K562/Adr200 and K562/Adr500 were generated by culturing K562 cells (CLS Cell Lines Service GmbH, Eppelheim, Germany) in step-wise incremental doses of Adr, ranging between 0.002 and 0.5 μM over a period of 2 months at 37°C with 5% CO 2 in a humidified incubator.

Techniques: Activity Assay, Expressing, Incubation, Western Blot, Binding Assay

Journal: Cells

Article Title: The Bibliometric Landscape of Gene Editing Innovation and Regulation in the Worldwide

doi: 10.3390/cells11172682

Figure Lengend Snippet: The research status and application fields of GE technologies (mainly CRISPR/Cas9).

Article Snippet: ZFNs , Srinivasan Chandrasegaran , Johns Hopkins University (JHU, USA) , 1996 , [ ] , Medicine and Health , K562, CD4 + T cells (X-linked severe combined immune deficiency (SCID)) , IL2RG , [ ] .

Techniques: CRISPR, TALENs, Infection, Functional Assay, Virus

Journal: Oncotarget

Article Title: Human HLA-A*02:01/CHM1 + allo-restricted T cell receptor transgenic CD8 + T Cells specifically inhibit Ewing sarcoma growth in vitro and in vivo

doi: 10.18632/oncotarget.9218

Figure Lengend Snippet: A. Flow cytometric determination of peptide specificity with specific CHM1- HLA-A*02:01-multimers shows CHM1 319 specificity of wild type CD8+ T cell clone CHM1-4B4. An irrelevant EZH2 666 /HLA-A*02:01-multimer served as control. B. CHM1-4B4 was only positive for the variable α-chain TRAV13-1*02, detected by the Vα8 primer and the variable β-chain TRBV13*01, detected by the Vβ23 primer and was thus considered clonal. All other sequenced PCR products were unspecific or showed no open reading-frame (dim bordered boxes). C. In IFNγ ELISpot analyses, clone CHM1-4B4 shows peptide specificity and HLA-A*02:01 restriction. A673 and TC71: HLA-A*02:01 + ES, SB-KMS-KS1: HLA-A*02:01 − ES, K562: MHC − NK cell control. E/T ratio for ELISpot assay: 1:4. Error bars represent standard deviation of triplicate experiments. Asterisks indicate significance levels.

Article Snippet: The MHC − erythroid leukemia cell line K562 was a gift from A. Knuth and E. Jäger (Krankenhaus Nordwest, Frankfurt, Germany).

Techniques: Control, Enzyme-linked Immunospot, Standard Deviation

Journal: Oncotarget

Article Title: Human HLA-A*02:01/CHM1 + allo-restricted T cell receptor transgenic CD8 + T Cells specifically inhibit Ewing sarcoma growth in vitro and in vivo

doi: 10.18632/oncotarget.9218

Figure Lengend Snippet: Specific reactivity against peptide-loaded T2 cells and several tumor cell lines was verified in 5A. IFNγ- and 5B. Granzyme-B ELISpot analyses (A673 and TC71: HLA-A*02:01 + ES, SB-KMS-KS1: HLA-A*02:01 − ES, K562: MHC − NK cell control,). Effector/target ratio for IFNγ-ELISpots is 1:4. Error bars represent standard deviation of triplicate experiments. Asterisks indicate significance levels; N.s., not significant.

Article Snippet: The MHC − erythroid leukemia cell line K562 was a gift from A. Knuth and E. Jäger (Krankenhaus Nordwest, Frankfurt, Germany).

Techniques: Enzyme-linked Immunospot, Control, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: RP11-115N4.1 overexpression significantly decreased K562 cell proliferation. (A) EdU assay for cell proliferation in K562 cells with RP11-115N4.1 overexpression. The percentage of EdU positive cells was detected by flow cytometry. The picture scale was 100 μ m. (B) Cell cycle assay of K562 cells with RP11-115N4.1 overexpression. (C) Cell apoptosis analysis in K562 cells with RP11-115N4.1 overexpression. **** P < 0.0001. Each experiment was repeated three times and results are means ± SD.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Over Expression, EdU Assay, Flow Cytometry, Cell Cycle Assay

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: RP11-115N4.1 promoted HSP70 transcription. (A) Volcano plot described that the differentially expressed mRNAs in K562 cells with RP11-115N4.1 overexpression. (B) GO analysis data described that the differentially expressed mRNAs in K562 cells with RP11-115N4.1 overexpression. (C, D) Validation of differentially expressed genes by qRT-PCR. * P < 0.05, ** P < 0.01. Each experiment was repeated three times and results are means ± SD.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Over Expression, Biomarker Discovery, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: RP11-115N4.1 promoted HSP70 expression by interacting with HNRNPH3. (A, B) RNA-pull-down assay to identify RP11-115N4.1 binding protein in K562 cells. The eluted proteins were separated by SDS-PAGE and subjected to silver staining. Antisense RNA to RP11-115N4.1 (AS) was used as a negative control. The red arrow indicates the band representing the RP11-115N4.1-specific binding protein identified by mass spectrometry as HNRNPH3. Western blot analysis confirmed that RP11-115N4.1 interacts with HNRNPH3. (C) RIP assay was performed using normal mouse IgG or the anti-HNRNPH3 antibody. GAPDH was used as the negative control. (D) Western blot of HSP70 and HNRNPH3 in K562 cells under different condition. β -Actin was used as an internal control. *** P < 0.001, **** P < 0.0001. Each experiment was repeated three times and results are means ± SD.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Expressing, Pull Down Assay, Binding Assay, SDS Page, Silver Staining, Negative Control, Mass Spectrometry, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: The supernatant of K562 cells overexpressed RP11-115N4.1 induced the inflammatory response of monocytes and inhibited the migration of trophoblast cells. (A) The level of HSP70 was determined by ELISA in K562 cell treated with RP11-115N4.1. (B) The level of HSP70 was determined by ELISA in the serum of URSA samples with high RP11-115N4.1 expression. (C) The levels of IL-6, IL-1β and TNF-α were determined by ELISA in human monocyte under different condition. LPS was used as a positive control (100 μg/mL). (D) Transwell assay in Swan 71 cell under different condition. The picture scale was 100 μ m. ** P < 0.01, *** P < 0.001, **** P < 0.0001. Each experiment was repeated three times and results are means ± SD.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Expressing, Positive Control, Transwell Assay

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: Schematic diagram of role of RP11-115N4.1. RP11-115N4.1 bound to HNRNPH3 and increased the protein level of HSP70 in K562 cells. HSP70 released outside the cell induced the upregulation of the inflammatory factor IL-6, IL-1 β and TNF- α of monocytes and inhibited the migration of Swan 71. The black dashed line represents an unknown mechanism.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Migration

Journal: Science Advances

Article Title: Bottom-up assembly of biomedical relevant fully synthetic extracellular vesicles

doi: 10.1126/sciadv.abg6666

Figure Lengend Snippet: ( A ) Schematic illustration of the fsEV formation process inside w/o droplet compartments produced by mechanical emulsification. miRNA-containing lipid vesicles are formed on the periphery of water droplets from SUV precursors for subsequent decoration with surface proteins. ( B ) Confocal microscopy images of fsEVs labeled with Liss Rhod PE–labeled fluorescent lipids (lipid membrane), Alexa Fluor 488–labeled CD9 (CD9), and Hoechst 33342–labeled miRNAs (hsa-miR-21, hsa-miR-124, hsa-miR-125, hsa-miR-126, hsa-miR-130, and hsa-miR-132). Scale bar, 2 μm. ( C ) Representative fluorescence confocal microscopy images of human dermal fibroblasts incubated for 24 hours with DiI (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate)–labeled natural EVs isolated from K562 media (left) or incubated for 24 hours with fluorescent fsEVs. Scale bars, 20 μm.

Article Snippet: K562 exosomes (HBM-K562) were obtained from HansaBioMed/Lonza, Switzerland.

Techniques: Produced, Emulsification, Confocal Microscopy, Labeling, Membrane, Fluorescence, Incubation, Isolation