mda231 cells  (Santa Cruz Biotechnology)

Journal: Oncology Reports

Article Title: Exploration of genes related to the development of cancer of unknown primary

doi: 10.3892/or.2025.8905

Figure Lengend Snippet: Migration assays were performed using the Boyden chamber method. After transfection of each siRNA into the (A) A549 and (B) MDA231 cells, the cells were incubated for 48 h before the migration assay. The cells that had migrated to the outer side of the membranes within 24 h were fixed and stained. A representative image of a triplicate analysis of each siRNA is shown. Scale bar, 20 µm. siRNA, small interfering RNA; Ctrl, control siRNA; SERF2, small EDRK-rich factor 2; PRKDC, DNA-dependent protein kinase catalytic subunit; S100A6, S100 calcium-binding protein A6; SH3GLB1, SH3-domain GRB2-like endophilin B1; CAPNS1, calpain, small subunit 1; PSMB4, proteasome subunit β type-4; RPL11, ribosomal protein L11; RPS7, ribosomal protein S7.

Article Snippet: The A549 human lung cancer cell line (cat. no. CCL-185) and the MDA231 breast cancer cell line (cat. no. HTB-26) were procured from American Type Culture Collection, and cultured in DMEM (Thermo Fisher Scientific, Inc.) with 10% FBS (MilliporeSigma) in accordance with the instructions provided by the suppliers.

Techniques: Migration, Transfection, Incubation, Staining, Small Interfering RNA, Control, Binding Assay

Journal: Oncology Reports

Article Title: Exploration of genes related to the development of cancer of unknown primary

doi: 10.3892/or.2025.8905

Figure Lengend Snippet: Western blotting showing the reduction in protein expression following siRNA-induced knockdown of PRKDC and PSMB4. (Top) Protein expression was reduced after transfection of the cells with siRNA for PRKDC (left) and PSMB4 (right) in both the A549 and MDA231 cells. β-actin was used as the loading control, whereas si-NC was used as the negative control. (Bottom) Relative reductions in PRKDC or PSMB4 protein expression were determined using western blot analysis conducted in triplicate. Data are presented as the mean and standard deviation. *P<0.05; **P<0.01. siRNA/si, small interfering RNA; si-NC, siRNA universal negative control; PRKDC, DNA-dependent protein kinase catalytic subunit; PSMB4, proteasome subunit β type-4.

Article Snippet: The A549 human lung cancer cell line (cat. no. CCL-185) and the MDA231 breast cancer cell line (cat. no. HTB-26) were procured from American Type Culture Collection, and cultured in DMEM (Thermo Fisher Scientific, Inc.) with 10% FBS (MilliporeSigma) in accordance with the instructions provided by the suppliers.

Techniques: Western Blot, Expressing, Knockdown, Transfection, Control, Negative Control, Standard Deviation, Small Interfering RNA

Journal: Signal transduction and targeted therapy

Article Title: Simultaneously targeting extracellular vesicle trafficking and TGF-β receptor kinase activity blocks signaling hyperactivation and metastasis.

doi: 10.1038/s41392-023-01711-1

Figure Lengend Snippet: Fig. 3 Rab27a knockdown decreases sEV secretion and inhibits the TGF-β signaling activity in breast cancer cells in vitro. sEVs were isolated from parental (P) MDA231 and MDA.Rab27a.shRNA (RAB27A knockdown (KD)) cell culture conditioned medium. a sEV secretion was quantified by BCA assay. b, c Particle size distribution and concentration evaluated by NTA. d Alix and TSG101 expression assessed by western blot in (P) and (KD) whole cell lysates (WCL) and sEVs. e, f pSMAD2 levels evaluated in (P) and (KD) cells treated (e) with 2 ng/mL rhTGF-β1 (increasing time intervals) or (f) with indicated rhTGF-β1 concentrations (8 h). Total (t)SMAD2 and β-Actin were used as loading controls. g, h The TGF-β/SMAD3 signaling reporter (Ad-CAGA-Fluc) activity was quantified in parental and RAB27A KD cells treated ± rhTGF-β1 for (g) 24 h or (h) 48 h. Results normalized by Gaussia luciferase (Ad-CMV-Gluc) activity. Results represent mean ± SD (n ≥3). Unpaired Student’s t-test used for comparison. **p < 0.01, ***p < 0.001

Article Snippet: Transient Rab27a knockdown in MDA231 cells was established by Rab27a siRNA (sc-41834, Santa Cruz Biotechnology, Bio-Strategy Pty Limited, Australia) transfection using LipofectamineTM RNAiMAX Transfection Reagent (Invitrogen, Thermo Fisher Scientific, Australia PTY LTD).

Techniques: Knockdown, Activity Assay, In Vitro, Isolation, shRNA, Cell Culture, BIA-KA, Concentration Assay, Expressing, Western Blot, Luciferase, Comparison

Journal: Signal transduction and targeted therapy

Article Title: Simultaneously targeting extracellular vesicle trafficking and TGF-β receptor kinase activity blocks signaling hyperactivation and metastasis.

doi: 10.1038/s41392-023-01711-1

Figure Lengend Snippet: Fig. 4 TGF-β signaling is inhibited by DMA, Heparin and PNP-Xyl in vitro. a–d sEVs were isolated from MDA231 cell culture conditioned medium after treatment ± 100 µM DMA (2 h). a sEV secretion quantified by BCA assay. b, c Particle size distribution and concentration evaluated by NTA. d pSMAD2 levels were assessed by western blot in MDA231 cells treated ± DMA (100 µM) and rhTGF-β1 (5 ng/mL). Total (t) SMAD2 and β-actin were used as loading controls. e, f TGF-β/SMAD3 signaling reporter (Ad-CAGA-Fluc) activity quantified in (e) MDA231 and (g) MCF7 cells treated ± rhTGF-β1 (5 ng/mL) ± DMA (24 h). Luciferase assay results normalized by Gaussia luciferase (Ad-CMV-Gluc) activity. g pSMAD2 levels assessed in MDA231 cells treated with rhTGF-β1 or MDA231-sEVs after treatment ± heparin. h, i TGF-β/SMAD3 signaling activity quantified in (h) MDA231 and (i) MCF7 cells treated with MDA231-sEVs ± heparin as in (e, f). j pSMAD2 levels assessed in MDA231 cells treated with rhTGF-β1 or MDA231-sEVs after treatment ± PNP-Xyl as in (d, g). k, l TGF-β/SMAD3 signaling activity quantified in (k) MDA231 and (l) MCF7 cells treated with MDA231-sEVs after treatment ± PNP-Xyl (24 h). Results represent mean ± SD (n ≥3). Unpaired Student’s t-test was used to analyze data in (a, c). One-way ANOVA test followed by Dunn’s Multiple Comparison test was used to analyze data in (e–f, h–i, k–l). ns: statistically non-significant, **p < 0.01, ***p < 0.001

Article Snippet: Transient Rab27a knockdown in MDA231 cells was established by Rab27a siRNA (sc-41834, Santa Cruz Biotechnology, Bio-Strategy Pty Limited, Australia) transfection using LipofectamineTM RNAiMAX Transfection Reagent (Invitrogen, Thermo Fisher Scientific, Australia PTY LTD).

Techniques: In Vitro, Isolation, Cell Culture, BIA-KA, Concentration Assay, Western Blot, Activity Assay, Luciferase, Comparison

Journal: Signal transduction and targeted therapy

Article Title: Simultaneously targeting extracellular vesicle trafficking and TGF-β receptor kinase activity blocks signaling hyperactivation and metastasis.

doi: 10.1038/s41392-023-01711-1

Figure Lengend Snippet: Fig. 5 sEVs induce cancer cell EMT, migration, and invasion in vitro. a MCF7 cell morphology evaluated by phase contrast after treatment ± rhTGF-β1 or MDA231-sEVs (5 days). Red arrows indicate elongated cells. Scale bar is equal to 100 µm. b ZO-1 and E-cadherin localization analyzed by immunofluorescence staining after single treatment ± rhTGF-β1 or MDA231-sEVs (5 days). Scale bar is equal to 50 µm (c) ZO-1 and E-cadherin expression assessed by western blot in MCF7 cells treated once ± rhTGF-β1 or MDA231-sEVs (5 days). β-actin used as loading control. d MCF7 cell invasion quantified in cells seeded in Matrigel-coated transwell inserts and treated ± rhTGF-β1 or MDA231-sEVs (48 h). Scale bar is equal to 100 µm. e, f Cell migration quantified by wound healing assay in (e) MDA231 and (f) MCF7 cell cultures infected ± Ad-CMV-Flag-SMAD7 and treated as indicated. Ad-CMV-GFP: control adenovirus. Results represent mean ± SD (n ≥3). One-Way ANOVA followed by Dunn’s Multiple Comparison Test. *p < 0.05, **p < 0.01, ***p < 0.001, ns: statistically non-significant

Article Snippet: Transient Rab27a knockdown in MDA231 cells was established by Rab27a siRNA (sc-41834, Santa Cruz Biotechnology, Bio-Strategy Pty Limited, Australia) transfection using LipofectamineTM RNAiMAX Transfection Reagent (Invitrogen, Thermo Fisher Scientific, Australia PTY LTD).

Techniques: Migration, In Vitro, Staining, Expressing, Western Blot, Control, Wound Healing Assay, Infection, Comparison

Journal: Signal transduction and targeted therapy

Article Title: Simultaneously targeting extracellular vesicle trafficking and TGF-β receptor kinase activity blocks signaling hyperactivation and metastasis.

doi: 10.1038/s41392-023-01711-1

Figure Lengend Snippet: Fig. 6 sEVs increase TGF-β signaling and enhance MDA231 breast cancer progression in vivo. a Illustration of breast cancer mouse model showing the orthotopic implantation of MDA231 cells. Unlabeled MDA231 and Gaussia luciferase-labeled MDA.Gluc cells were implanted contralaterally. b Experiment timeline for (a). c TGF-β/SMAD3 signaling reporter (Ad-CAGA-Fluc) activity quantified in MDA.Gluc tumors (3 animals/group) by IVIS. d–f Gaussia luciferase activity quantified by luciferase assay in (d) blood, (e) lung, and (f) bone samples (5–6 mice/ group). Animals are color-coded. Black dashed lines indicate the background activity for the Gaussia luciferase quantified in samples from non-implanted mice (n = 2). Results represent mean ± SEM. One-Way ANOVA followed by Tukey’s Multiple Comparison Test. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Transient Rab27a knockdown in MDA231 cells was established by Rab27a siRNA (sc-41834, Santa Cruz Biotechnology, Bio-Strategy Pty Limited, Australia) transfection using LipofectamineTM RNAiMAX Transfection Reagent (Invitrogen, Thermo Fisher Scientific, Australia PTY LTD).

Techniques: In Vivo, Luciferase, Labeling, Activity Assay, Comparison

Journal: Signal transduction and targeted therapy

Article Title: Simultaneously targeting extracellular vesicle trafficking and TGF-β receptor kinase activity blocks signaling hyperactivation and metastasis.

doi: 10.1038/s41392-023-01711-1

Figure Lengend Snippet: Fig. 8 In vivo TGF-β signaling activity and breast cancer progression are impaired by combined treatment with DMA and SB431542 at suboptimal doses. a Illustration of breast cancer mouse model showing MDA231 orthotopic implantation. Unlabeled MDA231 and Gaussia luciferase-labeled MDA.Gluc cells were implanted contralaterally. b Experiment timeline for (a). c TGF-β/SMAD3 signaling reporter (Ad-CAGA- Fluc) activity quantified in MDA.Gluc tumors (5–6 animals/group). d–g Gaussia luciferase activity quantified by luciferase assay in (d) blood, (e) lung, (f) bone, and (g) unlabeled MDA231 tumor samples (6 animals/group). Animals are color-coded. Red dashed lines indicate the background activity for the Gaussia luciferase quantified in samples from non-implanted mice (n = 2). One-Way ANOVA followed by Tukey’s Multiple Comparison Test. Unpaired Student’s t-test used for comparison of drug-treated groups. *p < 0.05, **p < 0.01, ***p < 0.001, ns: statistically non-significant

Article Snippet: Transient Rab27a knockdown in MDA231 cells was established by Rab27a siRNA (sc-41834, Santa Cruz Biotechnology, Bio-Strategy Pty Limited, Australia) transfection using LipofectamineTM RNAiMAX Transfection Reagent (Invitrogen, Thermo Fisher Scientific, Australia PTY LTD).

Techniques: In Vivo, Activity Assay, Luciferase, Labeling, Comparison