breast cancer cell line sum159  (Procell Inc)

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: a Schematic of the experimental setup with THP1 or Raw264.7 cells with indicated treatment. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p b Quantification of proportion of TREM2 in THP1 and Raw264.7 cells with PTX. n = 3 biological independent samples. c Western blot of THP1 and Raw264.7 cells with indicated treatments. The experiment was independently repeated three times with similar results. d Schematic of the experimental strategy. CM was collected from tumor cells with the indicated treatment. TREM2 expression in macrophages incubated with the CM was assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p e Quantification of proportion of TREM2 in THP1 incubated with CM of BT549 cells, SUM159 cells and MDA-MB-231 cells treated with PTX, in BMDM incubated with CM of Py8119 treated with PTX, in Raw264.7 incubated with CM from 4T1 and Py8119 cells treated with PTX, respectively. n = 3 biological independent samples. f Western blot analysis of TREM2 and related proteins in THP1, BMDMs, and Raw264.7 cells incubated with CM from BT549, SUM159, Py8119, and 4T1 cells, respectively. The experiment was independently repeated three times with similar results. g Representative immunofluorescence staining of THP1 cells incubated with BT549 CM. n = 3 biological independent samples. h Quantification of proportion of TREM2 in THP1 incubated with CM from BT549, SUM159, and MDA-MB-231 cells treated with Nab-PTX, respectively. n = 3 biological independent samples. i Quantification of the proportion of TREM2 in BMDM incubated with CM of Py8119 treated with Nab-PTX. n = 3 biological independent samples. j Cytokine array analysis of CM from BT549 cells treated indicated treatment. k Western blot analysis of the indicated proteins in Raw264.7 cells and BMDMs treated with recombinant FGF2. The experiment was independently repeated three times with similar results. l Quantification of the proportion of TREM2 in Raw264.7 cells and BMDMs treated with recombinant FGF2. n = 3 biological independent samples. m Schematic of the experimental strategy. CM was collected from tumor cells with the indicated treatment, and then pretreatment using an FGF2 neutralizing antibody. TREM2 expression in macrophages incubated with the CM was assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p n Western blot analysis of the indicated proteins in Raw264.7 and BMDMs incubated with the indicated CM. The experiment was independently repeated three times with similar results. o Quantification of the proportion of TREM2 in Raw264.7 and BMDMs incubated with the indicated CM. n = 3 biological independent samples. p Representative multiplex immunofluorescence staining of CD68, TREM2 and FGF2 in tumors treated with PTX or Nab-PTX ( n = 3 mice per group). The experiment was independently repeated three times with similar results. Data are shown as means ± S.D. and were analyzed by two-sided unpaired Student’s t test ( b , e , h and i ) and two-sided one-way ANOVA followed by Tukey’s test ( l and o ). Source data are provided as a Source Data file.

Article Snippet: Human embryonic kidney cell line HEK293T (ATCC, CRL-3216), human breast cancer cell line SUM159 (CLS Cell Lines service, 305116), mouse melanoma cell lines B16-F10 (ATCC, CRL-6475) and mouse macrophage cell line Raw264.7 (ATCC, TIB-71) were cultured in Dulbecco’s Modified Eagle Medium (DMEM).

Techniques: Western Blot, Expressing, Incubation, Immunofluorescence, Staining, Recombinant, Multiplex Assay

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: a Overlap of GeneCards and EDCODE public databases analyses to predict transcription factors that regulate TREM2. b Western blot analysis of the indicated proteins in BMDMs and Raw264.7 cells incubated with FGF2 (left) and with CM from Py8119 cells treated with PTX (right). The experiment was independently repeated three times with similar results. c Western blot analysis of the indicated proteins in Raw264.7 cells incubated with the indicated treatment. The experiment was independently repeated three times with similar results. d qPCR analysis of Trem2 expression in BMDMs transfected with Egr1 -expressing vectors. n = 3 biological independent samples. e Western blot analysis of TREM2 expression in BMDMs transfected with Egr1 -expressing vectors. The experiment was independently repeated three times with similar results. f Luciferase activity of HEK293T cells transfected with the indicated reporters and EGR1 -expressing or control vectors. n = 3 biological independent samples. g Abundance of EGR1 bound to the TREM2 promoter in BMDMs assessed by ChIP-qPCR. n = 3 biological independent samples. h Schematic of the Transwell assay. The first CM was collected from tumor cells with indicated treatment, and the second CM was collected from macrophages incubated with the first CM. The migration and invasion capabilities of macrophages incubated with the first CM were assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p i Quantification of migration and invasion of Py8119 cells induced by BMDM CM incubated with CM from Py8119 cells treated with PTX, and BT549, SUM159, and MDA-MB-231 cells induced by THP1 CM incubated with CM from BT549, SUM159, and MDA-MB-231 cells treated with PTX (left) or Nab-PTX (right), respectively. n = 5 biological independent samples. j Cytokine array analysis of CM from BMDMs with or without TREM2. k Quantification of migration and invasion of Py8119 cells incubated with indicated proteins. n = 3 biological independent samples. l Quantification of migration and invasion of Py8119 cells induced by CM from BMDMs ( Trem2 +/+ ) with CDE-096. n = 3 biological independent samples. m Western blot analysis of EMT- stimulating proteins in BMDMs incubated with indicated proteins. The experiment was independently repeated three times with similar results. n Schematic illustration showing that FGF2 promotes ERK1/2 phosphorylation to upregulate EGR1, which increases TREM2 expression in macrophages. Upregulated TREM2 enhances the secretion of Serpin E1, HGF, CCL3, and CXCL2 from macrophages to tumor cells, facilitating tumor metastasis via EMT. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p . Data are shown as means ± S.D. and were analyzed by two-sided unpaired Student’s t test ( d, f, i and l ), two-sided one-way ANOVA followed by Tukey’s test ( k ) and two-sided two-way ANOVA followed by Šídák’s test ( g ). Source data are provided as a Source Data file.

Article Snippet: Human embryonic kidney cell line HEK293T (ATCC, CRL-3216), human breast cancer cell line SUM159 (CLS Cell Lines service, 305116), mouse melanoma cell lines B16-F10 (ATCC, CRL-6475) and mouse macrophage cell line Raw264.7 (ATCC, TIB-71) were cultured in Dulbecco’s Modified Eagle Medium (DMEM).

Techniques: Western Blot, Incubation, Expressing, Transfection, Luciferase, Activity Assay, Control, ChIP-qPCR, Transwell Assay, Migration, Phospho-proteomics

Journal: eLife

Article Title: Synergistic effect of inhibiting CHK2 and DNA replication on cancer cell growth

doi: 10.7554/eLife.104718

Figure Lengend Snippet:

Article Snippet: Cell line ( H. sapiens ) , SUM159 , SUM159PT , Asterand Bioscience , Triple-negative breast cancer cell line.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Multi-Level Control of the ATM/ATR-CHK1 Axis by the Transcription Factor E4F1 in Triple-Negative Breast Cancer

doi: 10.3390/ijms23169217

Figure Lengend Snippet: Depletion of the transcription factor E4F1 sensitizes SUM159 Human Triple-Negative Breast Cancer (TNBC) cell line to the chemotherapy drugs Gemcitabine and Cisplatin. ( A )—Depletion of E4F1 in SUM159. Immunoblot analysis of E4F1 protein level in protein extracts prepared from SUM159 cells treated with either a control shRNA (sh Ctrl ) or a shRNA directed against E4F1 (sh E4F1 ), or with a combination of the latter sh E4F1 with an expression vector encoding an E4F1 cDNA resistant to this sh E4F1 (pE4F1*). Extracts were prepared 4 days after cell infection with lentiviral vectors expressing shRNAs. shRNA directed against human E4F1 have been previously validated ; ( B , C )—E4F1-depleted SUM159 cells fail to arrest in S-Phase upon Gemcitabine treatment. Three days after sh Ctrl or sh E4F1 treatment, cells were exposed to sub-lethal dose of Gemcitabine (140 nM, IC30) for 24 h; ( B ) Representative flow cytometry analysis (PI staining) of cell cycle distribution in populations of cells collected at 24 h after start of Gemcitabine treatment. Of note FACScan gating settings were meant to exclude sub-G1 cells; ( C ) Quantitation of the distribution of cells in G2/M, S and G0/G1 cell cycle phases was performed with the FlowJo software. Results are expressed as mean values ± SD of three independent experiments. ** p < 0.01, by a one way ANOVA test performed on GraphPad Prism; ( D – F) —shRNA-mediated depletion of E4F1 sensitizes TNBC SUM159 cells to cytotoxic drugs, an effect rescued by the co-expression of the sh E4F1 -resistant pE4F1* construct (see also on another TNBC cell line, HCC38). Three days after sh Ctrl , sh E4F1 or sh E4F1 + pE4F1* transduction, cells were exposed to sub-lethal doses of Gemcitabine (140 nM, IC30) or Cisplatin (1.5 mM) for 24 h and assessed for cell death by ( D ) Phase contrast microscopy, ( E , F ) Flow cytometry analysis for apoptosis /necrosis by staining dying cells with annexin V/7-AAD and propidium Iodide (PI). ( E ) Representative flow cytometry profile of annexin V/7-AAD/PI –positive cells, and ( F ) quantitation (FlowJo software) of three independent experiments expressed as mean values ± SD of three independent experiments. ** p < 0.01, *** p < 0.001 as determined by a one way ANOVA test performed on GraphPad Prism.

Article Snippet: Cell culture and treatments The SUM159 Triple-Negative Breast Cancer (TNBC) cell line was obtained from Asterand Bioscience, UK, and grown in Ham’s F-12 medium (Gibco), supplemented with 5% fetal bovine serum, 10 μg/mL insulin, 1 μg/mL hydrocortisone, 100 μg/mL streptomycin and 100 units/mL penicillin and treated 24 h at the IC30 of Gemcitabine (140 nM, Sandoz), or Cisplatin (1.5 μM, Sigma Aldrich, St. Louis, MO, USA).

Techniques: Western Blot, Control, shRNA, Expressing, Plasmid Preparation, Infection, Flow Cytometry, Staining, Quantitation Assay, Software, Construct, Transduction, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Multi-Level Control of the ATM/ATR-CHK1 Axis by the Transcription Factor E4F1 in Triple-Negative Breast Cancer

doi: 10.3390/ijms23169217

Figure Lengend Snippet: E4F1 directly controls a limited set of genes in the TNBC cell line SUM159, including a sub-program involved in the ATM/ATR-CHEK signaling pathway. ( A )—Fold change plot of differentially expressed genes, assessed by RNA-Seq, in SUM159 cells four days after the initial exposure to either sh Ctrl or sh E4F1 . The expression of a large set of genes is moderately deregulated in E4F1-depleted cells compared to control; ( B )—Identification of E4F1 target genes by ChIP-Seq. E4F1 ChIP-Seq read densities in the 3 kb regions surrounding the Transcription Start Sites (TSS) of the closest genes located nearby E4F1 binding sites, as identified in SUM159 cells. Out of the 116 ChIP-Seq peaks identified by ChIP-Seq in SUM159 cells ( n = 2; , 102 were in these 3kb regions and 94 in promoters, indicating that E4F1 is mainly located nearby or on the TSS of its target genes; ( C )—MEME logo sequence analysis of the DNA fragments bound by E4F1 in SUM159 cells reveals a human E4F1 consensus motif that is similar to the Mouse E4F1 consensus motif previously identified by our laboratory ; ( D )—Venn diagram of the overlap between all of the E4F1 bound genes identified by ChIP-Seq in SUM159 cells (GSE128104 (in red)) and genes differentially expressed ( p < 0.05) between SUM159 cells treated with either sh Ctrl or sh E4F1 (GSE128099 (in blue)). These genome-wide analyses were performed at a time where all of the cells treated with shRNAs were still viable (annexin-V negative) and still actively growing. The overlap defines a set of 57 genes directly bound and regulated by E4F1 in SUM159; ( E )—List of the 57 E4F1 direct target genes as defined in panel D. Table provides information about the differential expression of transcripts in SUM159 cells treated with either sh Ctrl or sh E4F1 (Ratio KO/WT), the distance of the E4F1 consensus motif to the transcription start site (TSS) of the gene, and whether these genes code for factors involved in cellular stress response and checkpoints (red+) including proteins involved in the ATM/ATR-CHK kinases axis (CHK1, PPP5C and TTI2).

Article Snippet: Cell culture and treatments The SUM159 Triple-Negative Breast Cancer (TNBC) cell line was obtained from Asterand Bioscience, UK, and grown in Ham’s F-12 medium (Gibco), supplemented with 5% fetal bovine serum, 10 μg/mL insulin, 1 μg/mL hydrocortisone, 100 μg/mL streptomycin and 100 units/mL penicillin and treated 24 h at the IC30 of Gemcitabine (140 nM, Sandoz), or Cisplatin (1.5 μM, Sigma Aldrich, St. Louis, MO, USA).

Techniques: RNA Sequencing, Expressing, Control, ChIP-sequencing, Binding Assay, Sequencing, Genome Wide, Quantitative Proteomics

Journal: International Journal of Molecular Sciences

Article Title: Multi-Level Control of the ATM/ATR-CHK1 Axis by the Transcription Factor E4F1 in Triple-Negative Breast Cancer

doi: 10.3390/ijms23169217

Figure Lengend Snippet: E4F1 directly binds and controls the CHEK1 , PPP5C , TTI2 and E4F1 genes in the TNBC cell line SUM159 and in a primary TNBC tumor-derived xenograft (TNBC PDX). ( A )—ChIP-Seq read densities detected at the CHEK1 , PPP5C , TTI2 genes and at the E4F1 gene itself, in SUM159 cells using either anti-E4F1 or control antibodies. Tag densities were calculated using the HOMER Suite and visualized with CISGENOME browser; ( B )—ChIP-qPCR validation of the presence of E4F1 on the promoter region of CHEK1 , PPP5C , TTI2 and E4F1 genes in SUM159 cells, 5 days after their infection with lentiviral vectors expressing shRNAs directed against either E4F1 (sh E4F1 ) or control sequence (sh Ctrl ). The CHEK2 promoter region and an intergenic region (NC3) are used as controls. Enrichments are represented as percentage of input (data are means ± SD, n = 3 independent experiments); ( C )—ChIP-qPCR analysis of the presence of E4F1 protein on the promoter region of CHEK1 , PPP5C , TTI2 and E4F1 genes in primary Human Triple-Negative Breast tumor-derived xenograft . ChIP-qPCR analyses were performed on freshly isolated tumor samples, two weeks after their engraftment on Nude mice. Enrichments are represented as percentage of input (data are means ± SD, n = 3 distinct tumors); ( D )—mRNA levels of E4F1 target genes coding for CHEK1, PPP5C and TTI2 in SUM159 cells treated with either shCtrl or sh E4F1 . Measured by RT-qPCR, 5 days after cell infection with lentiviral vectors expressing shRNAs. Data are means ± SD, n = 3 independent experiments. NS: non significative, ** p < 0.01, *** p < 0.001 as determined by a one way ANOVA test performed on GraphPad Prism.

Article Snippet: Cell culture and treatments The SUM159 Triple-Negative Breast Cancer (TNBC) cell line was obtained from Asterand Bioscience, UK, and grown in Ham’s F-12 medium (Gibco), supplemented with 5% fetal bovine serum, 10 μg/mL insulin, 1 μg/mL hydrocortisone, 100 μg/mL streptomycin and 100 units/mL penicillin and treated 24 h at the IC30 of Gemcitabine (140 nM, Sandoz), or Cisplatin (1.5 μM, Sigma Aldrich, St. Louis, MO, USA).

Techniques: Derivative Assay, ChIP-sequencing, Control, ChIP-qPCR, Biomarker Discovery, Infection, Expressing, Sequencing, Isolation, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Multi-Level Control of the ATM/ATR-CHK1 Axis by the Transcription Factor E4F1 in Triple-Negative Breast Cancer

doi: 10.3390/ijms23169217

Figure Lengend Snippet: The transcriptional control exercised by E4F1 on the TTT complex component TTI2, strongly impacts on ATM and ATR protein levels in the TNBC cell line, SUM159. ( A )—E4F1 depletion impacts on TTI2, ATM and ATR protein levels. Immunoblot analysis of indicated proteins in total extracts prepared from SUM159 cells treated with either sh Ctrl or sh E4F1 . Extracts were prepared 4 days after cell infection with lentiviral vectors expressing shRNAs; ( B )—E4F1 depletion impacts on TTI2 mRNA level, but not on ATM and ATR mRNA levels, nor on the mRNA level of TTI1, another component of the TTT complex. Measured by RT-qPCR, 5 days after cell infection with lentiviral vectors expressing either sh Ctrl or sh E4F1 . Data are means ± SD, n = 3 independent experiments. ** p < 0.01, as determined by a one way ANOVA test performed on GraphPad Prism; ( C )—siRNA-mediated depletion of TTI2 impacts on ATR and ATM protein levels in SUM159. Immunoblot analysis of indicated proteins in total extracts prepared from SUM159 cells treated for 48 h with either a control siRNA (siCtrl) or an siRNA directed against human TTI2 mRNA; ( D )—Immunoblot analysis of indicated proteins in total extracts prepared from SUM159 cells treated with either sh Ctrl or sh E4F1 and with the lysosomal inhibitor E64 (10 μM).

Article Snippet: Cell culture and treatments The SUM159 Triple-Negative Breast Cancer (TNBC) cell line was obtained from Asterand Bioscience, UK, and grown in Ham’s F-12 medium (Gibco), supplemented with 5% fetal bovine serum, 10 μg/mL insulin, 1 μg/mL hydrocortisone, 100 μg/mL streptomycin and 100 units/mL penicillin and treated 24 h at the IC30 of Gemcitabine (140 nM, Sandoz), or Cisplatin (1.5 μM, Sigma Aldrich, St. Louis, MO, USA).

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

Journal: International Journal of Molecular Sciences

Article Title: Multi-Level Control of the ATM/ATR-CHK1 Axis by the Transcription Factor E4F1 in Triple-Negative Breast Cancer

doi: 10.3390/ijms23169217

Figure Lengend Snippet: E4F1-depletion impacts on ATM/ATR-CHK1 signaling in response to CT drugs Gemcitabine and Cisplatin. ( A )—ShRNA-mediated depletion of E4F1 in SUM159 TNBC cells impacts on CHK1 protein level and on the S 345 phosphorylation of CHK1 by ATM/ATR in response to Gemcitabine and Cisplatin. This effect was reversed by the co-expression of the shRNA resistant pE4F1* cDNA. Immunoblot analysis of indicated proteins or phosphor-proteins (CHK1 P ) in total extracts prepared from SUM159 cells transduced with either control shRNA (sh Ctrl ), sh E4F1 , or sh E4F1 + pE4F1*, and treated for 24 h with Gemcitabine or Cisplatin. See also immunoblots in showing a similar experiment in another TNBC cell line, HCC38; ( B , C ) Immunofluorescence staining of DNA (blue), phospho (S139)- γH2AX (green) and ATR (red) in SUM159 cells transduced with either sh Ctrl , sh E4F1 , or sh E4F1 + pE4F1*, and upon antibiotics selection, treated for 24 h with Gemcitabine; ( B ) Representative experiment; ( C ) Quantitation of three independent experiments expressed as mean values ± SD. *** p < 0.001 as determined by two-tailed unpaired t -test performed on Graph Pad Prism. See also for immunoblotting analysis of γH2AX phosphorylation in cell lysates.

Article Snippet: Cell culture and treatments The SUM159 Triple-Negative Breast Cancer (TNBC) cell line was obtained from Asterand Bioscience, UK, and grown in Ham’s F-12 medium (Gibco), supplemented with 5% fetal bovine serum, 10 μg/mL insulin, 1 μg/mL hydrocortisone, 100 μg/mL streptomycin and 100 units/mL penicillin and treated 24 h at the IC30 of Gemcitabine (140 nM, Sandoz), or Cisplatin (1.5 μM, Sigma Aldrich, St. Louis, MO, USA).

Techniques: shRNA, Phospho-proteomics, Expressing, Western Blot, Transduction, Control, Immunofluorescence, Staining, Selection, Quantitation Assay, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Multi-Level Control of the ATM/ATR-CHK1 Axis by the Transcription Factor E4F1 in Triple-Negative Breast Cancer

doi: 10.3390/ijms23169217

Figure Lengend Snippet: E4F1-depletion does not impact on the formation of 53BP1 foci in response to Gemcitabine, suggesting the early step of the DNA damage-response, upstream of the ATM/ATR-CHK1 signaling, is still functional in E4F1-depleted cells. A and B- Immunofluorescence staining of DNA (blue) and of 53BP1 protein (red) in SUM159 cells transduced with either control sh Ctrl or sh E4F1 and, upon antibiotics selection, treated for 24 h with Gemcitabine. ( A ) representative experiment; ( B ) Quantitation of independent experiments expressed as mean values ± SD of three independent experiments. *** p < 0.001 as determined by two-tailed unpaired t -test performed on Graph Pad Prism.

Article Snippet: Cell culture and treatments The SUM159 Triple-Negative Breast Cancer (TNBC) cell line was obtained from Asterand Bioscience, UK, and grown in Ham’s F-12 medium (Gibco), supplemented with 5% fetal bovine serum, 10 μg/mL insulin, 1 μg/mL hydrocortisone, 100 μg/mL streptomycin and 100 units/mL penicillin and treated 24 h at the IC30 of Gemcitabine (140 nM, Sandoz), or Cisplatin (1.5 μM, Sigma Aldrich, St. Louis, MO, USA).

Techniques: Functional Assay, Immunofluorescence, Staining, Transduction, Control, Selection, Quantitation Assay, Two Tailed Test