human normal breast epithelial cells mcf 10a  (ATCC)

Journal: Cell Communication and Signaling : CCS

Article Title: A pro-carcinogenic oral microbe internalized by breast cancer cells promotes mammary tumorigenesis

doi: 10.1186/s12964-025-02635-9

Figure Lengend Snippet: F. nucleatum induces DNA damage leading to upregulation of error prone NHEJ repair pathway. a GSEA analysis from bulk-RNA sequencing of HCC1806 xenografts with intravenous FN23726 inoculation showing activation of DNA repair pathways. b Heatmap showing relative expression of genes involved in DNA damage and repair in FN infected breast tumors. c , d Representative immunocytochemistry images showing HCC1937 ( c ) and MCF10A-BRCA+/- ( d ) P0 cells exposed to F. nucleatum FN23726 and stained with γH2AX and RAD51 antibodies, as indicated. DAPI is used to stain the nuclei. e Immunoblots showing the expression of DNA-PKc, XLF and Ku80 in MCF10A-BRCA+/- and HCC1937 cells exposed to F. nucleatum FN23726 for 3 h and P0 (passage 0) or P2 (passage 2) cells. f , g Representative immunocytochemistry images showing HCC1937 and MCF10A-BRCA+/- P0 ( f ) and P1 ( g ) cells exposed to F. nucleatum FN23726 and stained with pATM and pBRCA1 antibodies, as indicated. DAPI is used to stain the nuclei. (h-i) Representative immunocytochemistry images showing SUM149 ( h ) and HCC1937 ( i ) cells exposed to F. nucleatum FN23726 and stained with DAD1 and CDK4 antibodies, as indicated. DAPI is used to stain the nuclei.

Article Snippet: Breast cancer cell lines MCF7, HCC1806, HCC1937, SUM149, and normal breast epithelial cell line MCF10A were procured from the ATCC.

Techniques: RNA Sequencing, Activation Assay, Expressing, Infection, Immunocytochemistry, Staining, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: A pro-carcinogenic oral microbe internalized by breast cancer cells promotes mammary tumorigenesis

doi: 10.1186/s12964-025-02635-9

Figure Lengend Snippet: Cellular uptake and intracellular survival of F. nucleatum . a Graphs present flow cytometry results showing intracellular F. nucleatum after 4 h and 24 h of co-culturing with HCC1806, HCC1937 and MCF10A-BRCA+/- cells. b Representative photomicrographs showing the uptake of FM 1-43 FX tagged F. nucleatum in MCF10A-BRCA+/- cells. DAPI is used to stain the nuclei. Phalloidin (red) is utilized to stain actin filaments. c Representative photomicrographs showing the FM 1-43 FX tagged F. nucleatum co-cultured with MCF10A-BRCA+/- followed by immunostaining with anti- F. nucleatum antibody without membrane permeabilization. d Representative photomicrographs showing the intracellular survival of FM 1-43 FX tagged F. nucleatum in MCF10A-BRCA+/- cells after 2 passages

Article Snippet: Breast cancer cell lines MCF7, HCC1806, HCC1937, SUM149, and normal breast epithelial cell line MCF10A were procured from the ATCC.

Techniques: Flow Cytometry, Staining, Cell Culture, Immunostaining, Membrane

Journal: Cell Communication and Signaling : CCS

Article Title: A pro-carcinogenic oral microbe internalized by breast cancer cells promotes mammary tumorigenesis

doi: 10.1186/s12964-025-02635-9

Figure Lengend Snippet: MCF10A-BRCA+/- cells exhibit altered functional properties as well as reduced olaparib and doxorubicin responsiveness upon F. nucleatum exposure. ( a ) Representative images show anchorage-dependent colony formation of MCF10A-BRCA+/- cells exposed to F. nucleatum followed by one (P1) or three (P3) passages. ( b ) Representative images show anchorage-independent colony formation (soft-agar colony-formation) of MCF10A-BRCA+/- cells exposed to F. nucleatum . Bar graph shows quantitative representation of the area of soft-agar colonies. N=3, Between group comparison by student’s t test. ( c ) Representative images show spheroid-migration of MCF10A-BRCA+/- cells exposed to F. nucleatum FN23726. Bar graph shows quantitative representation of distance migrated by the cells. N=3, Between group comparison by student’s t test. ( d ) Representative immunocytochemistry images showing MCF10A-BRCA+/- cells exposed to F. nucleatum FN23726 and stained with γH2AX antibodies, as indicated. DAPI is used to stain the nuclei. ( e ) Representative images of mammospheres (primary, secondary and tertiary) formed by MCF10A-BRCA+/- cells exposed to F. nucleatum FN23726. ( f ) Bar graphs show the number and area of mammospheres formed in each group. N=3, Between group comparison by student’s t test. ( g ) Representative data for ALDH1 activity in MCF10A-BRCA+/- cells exposed to F. nucleatum FN23726. ( h ) Dose response curves showing viability of MCF10A-BRCA+/-, HCC1937 and SUM149 cells exposed to F. nucleatum FN23726 (P2), in response to varying concentrations of Olaparib and Doxorubicin. Viability of cells were determined by MTT assay at 48 hours in two independent experiments with 4 replicates per group in each experiment (N=8). Dose response curves of Sham vs FN-cocultured cells was compared using 2 way ANOVA

Article Snippet: Breast cancer cell lines MCF7, HCC1806, HCC1937, SUM149, and normal breast epithelial cell line MCF10A were procured from the ATCC.

Techniques: Functional Assay, Comparison, Migration, Immunocytochemistry, Staining, Activity Assay, MTT Assay