kms11 cell line  (JCRB Cell Bank)

Journal: Leukemia

Article Title: The long non-coding RNA CRNDE regulates growth of multiple myeloma cells via an effect on IL6 signalling

doi: 10.1038/s41375-020-01034-y

Figure Lengend Snippet: a CRNDE+/+ and CRNDEΔ/Δ KMS11 clones (4 per genotype) were pulse labelled with CellTracer dye, and dilution of the dye due to cell division was monitored by flow cytometry. Numbers indicate fold decrease of mean fluorescence intensity (MFI) relative to day 0 (p < 0.05 for genotype as a significant variable; for individual comparisons, ****p < 0.0001; 2-way ANOVA). b MTT proliferation assay of CRNDE+/+ clones (n = 4), CRNDE+/Δ clones (n = 4) and CRNDEΔ/Δ clones (n = 2). Absorbance (590 nm) was measured over 6 days (p < 0.01 for genotype as a significant variable; for individual comparisons *p < 0.05 and **p < 0.01; 2-way ANOVA). c Cell adhesion of CRNDE+/+ and CRNDEΔ/Δ clones. Points represent the mean of duplicate experiments for 5 CRNDE+/+ clones and 4 CRNDEΔ/Δ clones (*p < 0.05; Mann–Whitney). CRNDE+/+ and CRNDEΔ/Δ clones (replicate cultures on 3 clones per genotype) were cultured in complete medium containing Bortezomib (d) or Dexamethasone (e) at the concentrations indicated for 72 h and assessed for apoptosis by FACS. Shown is the percentage of live cells (AnnexinV− Propidium Iodide-) (p values for genotype as a significant variable are shown; 2-way ANOVA). f Percentage of KMS11 cells (hCD38+) in bone marrows of recipient NSG mice at 6 weeks after injection with CRNDE+/+ or CRNDEΔ/Δ clones (2 clones per genotype, each injected into 5 mice (**p < 0.01; Mann–Whitney)).

Article Snippet: MM cell lines KMS11 [ 19 ], RPMI 8226 [ 20 ], OPM2 [ 21 ] and NC1-H929 [ 22 ] as well as RAJI (Burkitt’s lymphoma) [ 23 ] cell lines were cultured in RPMI-1640 medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% (or 20% for KMS11) foetal calf serum (FCS), 100 U/ml penicillin, 100 μg/ml streptomycin and 2 mM L-glutamine.

Techniques: Clone Assay, Flow Cytometry, Fluorescence, Proliferation Assay, MANN-WHITNEY, Cell Culture, Injection

Journal: Leukemia

Article Title: The long non-coding RNA CRNDE regulates growth of multiple myeloma cells via an effect on IL6 signalling

doi: 10.1038/s41375-020-01034-y

Figure Lengend Snippet: a Microarray analysis of CRNDE KMS11 CRISPR clones (2 clones per genotype). Shown is the number of genes up- and down-regulated in each comparison (cutoffs of FC < −2 or FC > 2 and ANOVA p ≤ 0.01). b Unsupervised clustering based on genes differentially expressed (F test p ≤ 0.01). c Scatter plot representation of comparison of CRNDE+/+ and CRNDEΔ/Δ KMS11 cells. Genes up- or down-regulated in CRNDEΔ/Δ cells relative to wild type are shown in red and green respectively. The probes for the IL6R, CDH2 and BMP6 genes are indicated. d GSEA analysis showing the Normalised enrichment score for Hallmark and Biocarta genesets significantly enriched (cut-off FDR q < 0.05) in CRNDE+/+ (blue) or CRNDEΔ/Δ (red) KMS11 clones. e Analysis by ChIP of enrichment of H3K4me3 (left panel) and H3K27me3 (right panel) at the promoters of the IL6R, CDH2 and BMP6 genes in CRNDE+/+ and CRNDEΔ/Δ KMS11 clones. The GAPDH (expressed) and HOXC9 (not expressed in KMS11) promoters were used as controls. Replicate ChIP experiments were performed on two clones per genotype (*p < 0.05, **p < 0.01; student’s t test).

Article Snippet: MM cell lines KMS11 [ 19 ], RPMI 8226 [ 20 ], OPM2 [ 21 ] and NC1-H929 [ 22 ] as well as RAJI (Burkitt’s lymphoma) [ 23 ] cell lines were cultured in RPMI-1640 medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% (or 20% for KMS11) foetal calf serum (FCS), 100 U/ml penicillin, 100 μg/ml streptomycin and 2 mM L-glutamine.

Techniques: Microarray, CRISPR, Clone Assay, Comparison

Journal: Leukemia

Article Title: The long non-coding RNA CRNDE regulates growth of multiple myeloma cells via an effect on IL6 signalling

doi: 10.1038/s41375-020-01034-y

Figure Lengend Snippet: a Expression of CRNDEh and IL6R analysed by real time RT-PCR in CD138+ plasma cells isolated from bone marrow of newly diagnosed MM patients (n = 68). Expression (relative to ACTB) was normalised to the mean level observed in a control group of healthy individuals. Samples were stratified into quartiles based on expression of CRNDE (p < 0.05; ANOVA with post-test for linear trend). b Surface IL6R expression. Left panel: Representative flow cytometry staining of a CRNDE+/+ (blue) and CRNDEΔ/Δ (red) KMS11 clone with anti-IL6R-PE antibody or isotype control (grey). Right panel: Quantitation of mean fluorescence intensity of IL6R staining (n = 4 clones for each genotype) (*p < 0.05; Mann–Whitney). c IL6-mediated activation of STAT3. Left panel: Representative Western blot analysis of phosphorylated (Tyr705)-STAT3 and total STAT3 in two CRNDE+/+ and CRNDEΔ/Δ clones (serum starved) following treatment with or without IL6 (2 ng/ml) for 90 min. Right panel: Quantitation of Western blots. Shown is the ratio of phospho-STAT3: total STAT3 (n = 4 clones for each genotype) (**p < 0.01; ***p < 0.001; ANOVA). d IL6-mediated proliferation. Following pulse labelling with CellTracer dye, KMS11 clones (4 clones per genotype) were grown in minimum medium without (left) or with (right) 2 ng/ml IL6. Numbers indicate fold decrease in MFI relative to day 0. The p values for genotype as a significant factor are shown. For comparisons at each time point, *p < 0.05, ****p < 0.0001 (2-way ANOVA).

Article Snippet: MM cell lines KMS11 [ 19 ], RPMI 8226 [ 20 ], OPM2 [ 21 ] and NC1-H929 [ 22 ] as well as RAJI (Burkitt’s lymphoma) [ 23 ] cell lines were cultured in RPMI-1640 medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% (or 20% for KMS11) foetal calf serum (FCS), 100 U/ml penicillin, 100 μg/ml streptomycin and 2 mM L-glutamine.

Techniques: Expressing, Quantitative RT-PCR, Clinical Proteomics, Isolation, Control, Flow Cytometry, Staining, Quantitation Assay, Fluorescence, Clone Assay, MANN-WHITNEY, Activation Assay, Western Blot

Journal: BMC Cancer

Article Title: Targeting autophagy increases the efficacy of proteasome inhibitor treatment in multiple myeloma by induction of apoptosis and activation of JNK

doi: 10.1186/s12885-022-09775-y

Figure Lengend Snippet: IRE1α inhibitors in combination with ixazomib have synergistic effects on cytotoxicity of multiple myeloma cells under support of mesenchymal stromal cell line (hMSC-TERT). A KMS11 and RPMI-8226 cells were treated with 10 nM ixazomib alone or combined with 30 μM STF-083010 for five days with or without support of immortalized stromal cells (hMSC-TERT), ( n = 3). Cell viability were measured by PI staining. B Total RNA was extracted after 16 h of treatment as described above. B , C XBP1s , ATF6 and EIF2AK3 mRNA levels were assessed using RT-qPCR, ( n = 3). P values were calculated by two-way analysis of variance (ANOVA) for A, B and one-way analysis for C. A P value of less than 0.05 was considered statistically significant for all analyses. (* p < 0.05, ** p < 0.01, *** p < 0.001 versus control)

Article Snippet: Human cell lines KMS11 and RPMI-8226 were originally obtained from JCRB Cell Bank (Japanese Collection of Research Bank) and DSMZ (German Collection of Microorganisms and Cell Cultures), Braunschweig, Germany, respectively.

Techniques: Staining, Quantitative RT-PCR, Control

Journal: BMC Cancer

Article Title: Targeting autophagy increases the efficacy of proteasome inhibitor treatment in multiple myeloma by induction of apoptosis and activation of JNK

doi: 10.1186/s12885-022-09775-y

Figure Lengend Snippet: STF-083010 in combination with ixazomib initiates apoptosis in multiple myeloma cell lines. KMS11 and RPMI-8226 cells were treated as described above for 16 h, ( n = 3). A Using RT-qPCR, PUMA and NOXA mRNA expression levels were measured. P values were calculated by one-way analysis of variance (ANOVA). A P value of less than 0.05 was considered statistically significant for all of the analysis (* p < 0.05, ** p < 0.01, *** p < 0.001). B Protein levels of cleaved-PARP, PARP and β-actin as loading control were detected in KMS11 and RPMI-8226 using western blot

Article Snippet: Human cell lines KMS11 and RPMI-8226 were originally obtained from JCRB Cell Bank (Japanese Collection of Research Bank) and DSMZ (German Collection of Microorganisms and Cell Cultures), Braunschweig, Germany, respectively.

Techniques: Quantitative RT-PCR, Expressing, Control, Western Blot

Journal: BMC Cancer

Article Title: Targeting autophagy increases the efficacy of proteasome inhibitor treatment in multiple myeloma by induction of apoptosis and activation of JNK

doi: 10.1186/s12885-022-09775-y

Figure Lengend Snippet: Inhibition of autophagy in combination with ixazomib induces cell toxicity in multiple myeloma cells. KMS11 were treated with 10 nM ixazomib and in combination with autophagy inhibitors bafilomycin A1 (BAF, 5 nM) ( n = 3) or chloroquine (CQ, 40 μM), ( n = 2). A Cell viability was measured using PI staining after 72 h with BAF and CQ combined treatments respectively. B The table represents bliss formula calculation for drug combination analysis. The percentage of inhibitory effect of combined drugs was calculated by bliss formula in comparison with observed inhibitory effect. Based on this formula, we showed that ixazomib in combination with autophagy inhibitors (BAF or CQ) act synergistic. In addition, combination of Index was calculated based on Chou-Talalay. C Primary BM MNCs were isolated from multiple myeloma patients at diagnosis ( n = 7, one representative is shown) and treated with ixazomib 10 nM, chloroquine 40 μM and ixazomib plus chloroquine. After 48 h, 5 × 10. 4 of all pretreated cells were seeded in CFU culture medium. Colony numbers were counted after 14 days of culture using inverted light microscopy. P values were calculated using one-way ANOVA. A P value of less than 0.05 was considered statistically significant for all of the analysis (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: Human cell lines KMS11 and RPMI-8226 were originally obtained from JCRB Cell Bank (Japanese Collection of Research Bank) and DSMZ (German Collection of Microorganisms and Cell Cultures), Braunschweig, Germany, respectively.

Techniques: Inhibition, Staining, Comparison, Isolation, Biomarker Discovery, Light Microscopy

Journal: BMC Cancer

Article Title: Targeting autophagy increases the efficacy of proteasome inhibitor treatment in multiple myeloma by induction of apoptosis and activation of JNK

doi: 10.1186/s12885-022-09775-y

Figure Lengend Snippet: Autophagy inhibitors in combination with ixazomib induce apoptosis and arrest cell cycle at G 1 phase in multiple myeloma cells. KMS11 cells were treated with ixazomib and autophagy inhibitors as described above. A Apoptosis was measured by analysis of apoptosis fraction using annexin-V/PI staining. (BAF, 5 nM) ( n = 3) or chloroquine (CQ, 40 μM), ( n = 2). P values were calculated by one-way analysis of variance (ANOVA). A P value of less than 0.05 was considered statistically significant (* p < 0.05, ** p < 0.01, *** p < 0.001). B Using western blot protein levels of caspase3, cleaved-caspase3, PARP, cleaved PARP, p21 CIP1 and p27. KIP1 were measured. β-actin was usedas loading control, ( n = 3)

Article Snippet: Human cell lines KMS11 and RPMI-8226 were originally obtained from JCRB Cell Bank (Japanese Collection of Research Bank) and DSMZ (German Collection of Microorganisms and Cell Cultures), Braunschweig, Germany, respectively.

Techniques: Staining, Western Blot, Control

Journal: BMC Cancer

Article Title: Targeting autophagy increases the efficacy of proteasome inhibitor treatment in multiple myeloma by induction of apoptosis and activation of JNK

doi: 10.1186/s12885-022-09775-y

Figure Lengend Snippet: JNK inhibition rescues multiple myeloma cells from cell death mediated by combination treatment. KMS11 were treated with 10 nM ixazomib and in combination with autophagy inhibitors as described above, ( n = 3). A Phosphorylation levels of JNK-T 183 /Y. 185 and protein levels of JNK and BIM were measured using western blot with β-actin as a loading control. B KMS11 cells were treated with 10 nM ixazomib, 5 nM bafilomycin A1, 3 μM JNK inhibitor (JNK-IN-8) and combination treatment (ixa + baf and ixa + baf + JNK). Cell viability of KMS11 cells were measured using PI staining. P value were calculated by two-way analysis of variance (ANOVA). A p value of less than 0.05 was considered statistically significant (* p < 0.05, ** p < 0.01, *** p < 0.001 versus control)

Article Snippet: Human cell lines KMS11 and RPMI-8226 were originally obtained from JCRB Cell Bank (Japanese Collection of Research Bank) and DSMZ (German Collection of Microorganisms and Cell Cultures), Braunschweig, Germany, respectively.

Techniques: Inhibition, Phospho-proteomics, Western Blot, Control, Staining

Journal: Journal of Clinical Medicine

Article Title: Epigenetic Regulation of Mitochondrial Quality Control Genes in Multiple Myeloma: A Sequenom MassARRAY Pilot Investigation on HMCLs

doi: 10.3390/jcm10061295

Figure Lengend Snippet: Mean of DNA methylation values (and standard deviation) of CpG sites located within the analysed genes in HMCLs (MM1s, KMS11) and normal control cells (PBMCs). HMCLs, human myeloma cell lines; FDR: False Discovery Rate.

Article Snippet: MM1s and KMS11 human multiple myeloma cell lines (HMCLs) were obtained from the Leibniz Institute DSMZ (German Collection of Microorganisms and cell cultures GmbH) and the Japanese Collection of Research Bioresources (National Institute of Health Sciences Japan), respectively.

Techniques: DNA Methylation Assay, Standard Deviation, Control