human tnbc cell line bt549  (Procell Inc)

Journal: Cell Communication and Signaling : CCS

Article Title: Artificial intelligence-driven discovery of YH395A: A novel TGFβR1 inhibitor with potent anti-tumor activity against triple-negative breast cancer

doi: 10.1186/s12964-025-02337-2

Figure Lengend Snippet: YH395A inhibited breast cancer cell proliferation. (A) Breast cancer cell lines (MDA-MB-231, MDA-MB-468, BT549, SUM159, and BT-20), normal mammary epithelial cells (MCF-10 A), human gastric mucosal epithelial cells (GES-1), and human embryonic lung fibroblast cells (MRC-5) were treated with the indicated concentrations of YH395A or YR-290. After 96 h, SRB assay was performed. The bars indicate the mean ± SD. (B) Colony formation assays of breast cancer cells

Article Snippet: The MCF-10 A, GES-1, MRC-5, and triple-negative breast cancer (TNBC) cell lines BT549 (RRID: CVCL_0062), MDA-MB-468 (RRID: CVCL_0419), BT-20 (RRID: CVCL_0169), and MDA-MB-231(RRID: CVCL_0063) were sourced from the American Type Culture Collection (ATCC, USA), while SUM159 cells were generously donated by Dr. Yuzhu Zhang (Guangdong Academy of Chinese Medicine, China).

Techniques: Sulforhodamine B Assay

Journal: Narra J

Article Title: Dual sgRNA-directed knockout survivin gene expression using CRISPR/Cas9 technology for editing survivin gene in triple-negative breast cancer

doi: 10.52225/narra.v4i3.1177

Figure Lengend Snippet: Schematic diagram of survivin gene knockout using clustered regularly interspaced short palindromic repeats (CRISPR)-associated nuclease 9 (CRISPR/Cas9) in triple-negative breast cancer (TNBC) cell line BT549. EGFP: enhanced green fluorescent protein; PCR: polymerase chain reaction; sgRNA: single guide RNA

Article Snippet: TNBC cell line BT549 was obtained from the American Type Culture Collection (ATCC) by the Laboratory of Experimental Pathology at the University of Tsukuba, Tsukuba, Japan.

Techniques: Gene Knockout, CRISPR, Polymerase Chain Reaction

Journal: Narra J

Article Title: Dual sgRNA-directed knockout survivin gene expression using CRISPR/Cas9 technology for editing survivin gene in triple-negative breast cancer

doi: 10.52225/narra.v4i3.1177

Figure Lengend Snippet: Cleavage activity assay of single guide RNAs (sgRNAs) in BT549 cell line. Fluorescence intensity was quantified by using ImageJ software. Data was presented as mean±SD of fluorescence intensity ratio to cells without sgRNA transfection (control). *** Significantly different at p <0.001. A in the figure refers to sgRNA survin1/2 or survex A, while B refers to sgRNA survin1/2 or survex B.

Article Snippet: TNBC cell line BT549 was obtained from the American Type Culture Collection (ATCC) by the Laboratory of Experimental Pathology at the University of Tsukuba, Tsukuba, Japan.

Techniques: Activity Assay, Fluorescence, Software, Transfection, Control

Journal: Narra J

Article Title: Dual sgRNA-directed knockout survivin gene expression using CRISPR/Cas9 technology for editing survivin gene in triple-negative breast cancer

doi: 10.52225/narra.v4i3.1177

Figure Lengend Snippet: Establishment of survivin knockout in BT549 cells using single sgRNA. (A) PCR amplification of wild-type and 6 clones (#1–#6) in exon 2 of survivin gene targeted region. (B) Alignments of wild-type cells and all clones’ sequences using forward primer (top) and reverse primer (bottom). (C) Survivin protein expression level in wild-type cells and all clones. Data was presented as mean±SD. *** Significantly different at p <0.001, where each clone was compared to wild-type cells as control.

Article Snippet: TNBC cell line BT549 was obtained from the American Type Culture Collection (ATCC) by the Laboratory of Experimental Pathology at the University of Tsukuba, Tsukuba, Japan.

Techniques: Knock-Out, Amplification, Clone Assay, Expressing, Control

Journal: Narra J

Article Title: Dual sgRNA-directed knockout survivin gene expression using CRISPR/Cas9 technology for editing survivin gene in triple-negative breast cancer

doi: 10.52225/narra.v4i3.1177

Figure Lengend Snippet: Establishment of survivin knockout in BT549 cells using dual single guide RNAs (sgRNAs). (A) Schematic of primer design to detect wild-type and knockout of survivin gene. (B) Polymerase chain reaction (PCR) amplification of wild-type cells and six clones (#1–#6) in exon 2 of survivin gene targeted region. (C) Alignments of wild-type cells and all clones’ sequences show the success of survivin exon 2 deletion. A dash was used to symbolize the deletion of a single nucleotide. (D) Survivin protein expression level in wild-type cells and all clones. WT: wild type; KO: knockout; #1–#6: clone #1–#6.

Article Snippet: TNBC cell line BT549 was obtained from the American Type Culture Collection (ATCC) by the Laboratory of Experimental Pathology at the University of Tsukuba, Tsukuba, Japan.

Techniques: Knock-Out, Polymerase Chain Reaction, Amplification, Clone Assay, Expressing

Journal: Breast Cancer Research and Treatment

Article Title: Reciprocal regulation of forkhead box C1 and L1 cell adhesion molecule contributes to triple-negative breast cancer progression

doi: 10.1007/s10549-023-07177-7

Figure Lengend Snippet: FOXC1 is correlated with L1CAM, and its depletion leads to decreased L1CAM in TNBC cells. a Differences in L1CAM mRNA expression were examined between patients with a high ( n = 375) and low level ( n = 585) of FOXC1 mRNA. The mean was used for the cutoff value. b – c Two scatter plots were created to indicate the relation between FOXC1 and L1CAM mRNA expression in two public cohorts: TCGA and the Metastatic Breast Cancer Project, respectively (Pearson’s correlation coefficient r = 0.370, P < 0.001; r = 0.244, P = 0.002). d FOXC1 and L1CAM proteins were examined in four TNBC cell lines, with GAPDH serving as the loading control. e FOXC1 and L1CAM proteins were both down-regulated in BT549-shFOXC1 cells (by two different shFOXC1 sequences: A1 and A3) compared to BT549-shNC. β-Actin served as the loading control. f Real-time PCR detected decreased transcriptional levels of FOXC1 and L1CAM in two BT549-shFOXC1 cell clones: BT549-shFOXC1-A1 and BT549-shFOXC1-A3, compared to BT549-shNC. NC: BT549-shFOXC1-NC. A1 and A3: knockdown of FOXC1 with two shRNA sequences: RNA1 and RNA3 in BT549 cells, respectively. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The SUM149, HCC1937, MDA-MB-231 and BT549 TNBC cell lines were obtained from the ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction, Clone Assay, Knockdown, shRNA

Journal: Breast Cancer Research and Treatment

Article Title: Reciprocal regulation of forkhead box C1 and L1 cell adhesion molecule contributes to triple-negative breast cancer progression

doi: 10.1007/s10549-023-07177-7

Figure Lengend Snippet: Inhibition of L1CAM suppresses expression of FOXC1 proteins, cell proliferation, invasion and migration. a Down regulation of both L1CAM and FOXC1 proteins was detected in BT549-siL1CAM cells. GAPDH served as the loading control. b Real-time PCR detected a considerable decrease of L1CAM mRNA, but only a slight decrease of FOXC1 mRNA in BT549-siL1CAM cells. c CCK-8 assay showed L1CAM knockdown decreased the capacity of cell proliferation. d Cell migration and invasion were impaired after L1CAM knockdown in BT549 cells. e L1CAM knockdown suppressed BT549 cell migration in a wound healing assay. NC: negative control, siL1CAM: a small interfering RNA used to knockdown L1CAM mRNA in BT549 cells. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The SUM149, HCC1937, MDA-MB-231 and BT549 TNBC cell lines were obtained from the ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Inhibition, Expressing, Migration, Control, Real-time Polymerase Chain Reaction, CCK-8 Assay, Knockdown, Wound Healing Assay, Negative Control, Small Interfering RNA

Journal: Breast Cancer Research and Treatment

Article Title: Reciprocal regulation of forkhead box C1 and L1 cell adhesion molecule contributes to triple-negative breast cancer progression

doi: 10.1007/s10549-023-07177-7

Figure Lengend Snippet: Overexpression of L1CAM induces expression of FOXC1 proteins, cell proliferation, cell invasion and migration. a Western blotting showed L1CAM and FOXC1 proteins after overexpression of L1CAM in BT549-shFOXC1-A1 and BT549-shFOXC1-A3, i.e., BT549-shFOXC1-L1CAM-OA1 and BT549-shFOXC1-L1CAM-OA3. Their controls were represented as BT549-shFOXC1-NA1 and BT549-shFOXC1-NA3. β-Actin served as the loading control. b – c Real-time PCR showed the relative mRNA expression of L1CAM and FOXC1 after overexpressing L1CAM in BT549-shFOXC1 cells. d – e CCK-8 assays showed the effect of L1CAM overexpression on cell proliferation in BT549-shFOXC1-L1CAM. f – g Cell migration and invasion were evaluated after overexpressing L1CAM in BT549-shFOXC1 cells. h–i Upregulation of L1CAM in BT549-shFOXC1 cells enhanced cell migration in vitro wound healing assays. NA1: BT549-shFOXC1-A1 transfected with empty vector control. OA1: BT549-shFOXC1-A1 cells overexpressing L1CAM. NA3: BT549-shFOXC1-A3 transfected with empty vector control. OA3: BT549-shFOXC1-A3 cells overexpressing L1CAM. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The SUM149, HCC1937, MDA-MB-231 and BT549 TNBC cell lines were obtained from the ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Over Expression, Expressing, Migration, Western Blot, Control, Real-time Polymerase Chain Reaction, CCK-8 Assay, In Vitro, Transfection, Plasmid Preparation