hmcls mm1s  (ATCC)

Journal: Blood Advances

Article Title: MCT1 is a predictive marker for lenalidomide maintenance therapy in multiple myeloma

doi: 10.1182/bloodadvances.2021005532

Figure Lengend Snippet: Overexpression of MCT1 attenuates lenalidomide cytotoxicity in vitro and in vivo. (A-B) Cell proliferation analysis of MM1S (A) and U266 (B) cells, which were infected with lentivirus via control constructs (empty vector [EV]) or constructs to induce MCT1 or CD147 expression, treated with dimethyl sulfoxide (DMSO) or 10 µM lenalidomide for 72 h. (C) Quantification of tumor growth during lenalidomide or vehicle control treatment using caliper measurements (Ctrl, Len: n = 4 tumors). Xenograft tumors from U266 cells expressing MCT1 or the EV control. (D) Positron emission tomography (PET) images, using 18 F-fluorodeoxyglucose, of representative immunocompromised NOD-SCID mice bearing xenograft tumors expressing MCT1 or the EV control. Images were taken after 7 days of respective treatment. Arrows indicate tumors. The color scale indicates the percentage of injected dose per gram (% ID/g). Bars represent 10 mm. (E) Total lesion glycolysis of tumors determined by PET-MRI on day 7 after respective treatments. (F) Representative immunohistochemical analysis of tumors derived from the mice shown in panel D, with histomorphology visualized with hematoxylin-eosin staining and expression of cleaved caspase 3 (CC-3), MCT1, and CD79a. Analysis was performed on 10 high-power fields for each stain. Bars represent 100 µm. (G) Quantification of CC-3 staining shown in panel F. Data are expressed as means ± standard deviation. n.s., not significant; * P < .05, ** P < .01, by 1-sample t test or Student t test.

Article Snippet: The HMCLs MM1S (ATCC: CRL-2974) and U266 (DSMZ: ACC-9) cells were cultured in RPMI-1640 (Gibco) with 10% heat-inactivated fetal bovine serum and 1% penicillin-streptomycin.

Techniques: Over Expression, In Vitro, In Vivo, Infection, Control, Construct, Plasmid Preparation, Expressing, Positron Emission Tomography, Injection, Immunohistochemical staining, Derivative Assay, Staining, Standard Deviation

Journal: BMC Cancer

Article Title: MAFb protein confers intrinsic resistance to proteasome inhibitors in multiple myeloma

doi: 10.1186/s12885-018-4602-4

Figure Lengend Snippet: Expression of MAFb gene and protein in primary plasma cells and HMCLs. Affymetrix expression of MAFb mRNA was high in the GEP MF subgroup ( a ). Gene expression of 803 newly diagnosed MM patients was measured by U133 plus2.0 Affymetrix oligonucleotide microarray probe set 218559_s_at. The box plots indicate the middle of GEP signaling and dot lines with bars in black indicate quartiles. The cells were cultured in the growth mediate for 48 h and harvested for isolated RNA and MAFb mRNA in 27 HMCLs by qRT-PCR analysis as described in Material and methods ( b ). The cells were cultured in the growth mediate for 48 h and MAFb protein was measured in HMCLs by immunoblotting analysis described in Materials and Methods ( c ). MAFb protein in nuclei and cytoplasm of HMCLs ( d ) and MM PC from 4 patients ( e ) was determined with immunofluorescence staining with DAPI counterstaining. Images were taken with a fluorescence microscope with digital camera as described in supplemental data

Article Snippet: Human multiple myeloma cell lines (HMCLs) MM1S was purchased from ATCC (Manassas, VA, Catalogue number: CRL-2974).

Techniques: Expressing, Clinical Proteomics, Gene Expression, Microarray, Cell Culture, Isolation, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Fluorescence, Microscopy

Journal: BMC Cancer

Article Title: MAFb protein confers intrinsic resistance to proteasome inhibitors in multiple myeloma

doi: 10.1186/s12885-018-4602-4

Figure Lengend Snippet: High MAFb expression is associated with resistance to PIs. SACHI ( a ), H929 ( b ), EJM ( c ), XG2 ( d ), L363 ( e ), and OPM-2 ( f ) cells were seeded at 2 × 10^4 per well in 96-well plates in the presence of indicated concentrations of Bzb or CFZ for 48 h. Cell survival was measured by MTT Assay. Results are presented as mean ± SE ( n = 4). Data are representative of 3 separate experiments. HMCLs were treated with serial concentrations of Bzb (G) and CFZ (H) for 48 h and cell viability was measured by MTT assay. The IC50 of Bzb (G) and CFZ (H) were classified based on MAFb protein (+) or (−)

Article Snippet: Human multiple myeloma cell lines (HMCLs) MM1S was purchased from ATCC (Manassas, VA, Catalogue number: CRL-2974).

Techniques: Expressing, MTT Assay

Journal: BMC Cancer

Article Title: MAFb protein confers intrinsic resistance to proteasome inhibitors in multiple myeloma

doi: 10.1186/s12885-018-4602-4

Figure Lengend Snippet: Inhibition of GSK3 activity by SB216763 stabilized MAFb protein. HMCLs were treated with 5 μg/ml of CHX for serial indicated time points to inhibit de novo protein synthesis. The MAFb protein was determined by immunoblotting analysis using anti-MAFb. The membranes were striped and reblotted with Anti-β-actin ( a ). The half-life of MAFb protein was determined by autoradiographs analysis using Adobe Photoshop software and NIH image software ( b - f ). HMCLs were treated with or without a specific GSK3 inhibitor, SB216763, at a concentration of 5 μg/ml for indicated times. MAFb protein was determined by immunoblotting analysis using anti-MAFb antibody. The membranes were striped and reblotted with Anti-β-actin to indicate protein loading ( a ). The protein decay curve is as described in Fig. 3b (b-f)

Article Snippet: Human multiple myeloma cell lines (HMCLs) MM1S was purchased from ATCC (Manassas, VA, Catalogue number: CRL-2974).

Techniques: Inhibition, Activity Assay, Western Blot, Software, Concentration Assay