Journal: Cell & Bioscience

Article Title: MiR-181a targets STING to drive PARP inhibitor resistance in BRCA - mutated triple-negative breast cancer and ovarian cancer

doi: 10.1186/s13578-023-01151-y

Figure Lengend Snippet: MiR-181a overexpression leads to Olaparib resistance and downregulates STING. A – C Quantification by RT-qPCR of miR-181a levels in miR-181a-OV and empty vector (CTRL) MDA-MB-436 ( A ), HCC1395 ( B ), and HCC1937 ( C ) cell lines (Student’s t-test). D Western blotting analysis for STING and β-actin (loading control) comparing miR-181a-OV and empty vector (CTRL) in MDA-MB-436, HCC1395, and HCC1937 cell lines. E – G Drug sensitivity assays comparing miR-181a-OV and empty vector (CTRL) in MDA-MB-436 ( E ), HCC1395 ( F ), and HCC1937 ( G ) cell lines, treated with different concentrations of olaparib (Two-way ANOVA and Sidak’s multiple comparisons test). H Western blotting analysis for STING, TBK1, cGAS, and β-actin (loading control), comparing miR-181a-OV and empty vector (CTRL) HCC1937 cell lines with or without olaparib treatment. I Western blotting analysis for STING and β-actin (loading control) comparing parental and OlaR in MDA-MB-436, HCC1395, and HCC1937 cell lines. J Western blotting analysis for STING and β-actin (loading control) comparing parental and OlaR HCC1937 cell line in resting and olaparib-treated (24 and 48 h) conditions. K Western blotting analysis for STING and β-actin (loading control) comparing parental and OlaR HCC1937 cell line in resting and cisplatin-treated (12 and 24 h) conditions. L , M Representative images of STING IHC ( L ) and miR-181a ISH ( M ) for TNBC tumor tissues in the TMA. Images of cases 1 and 2 (STING low, miR-181a high) and cases 3 and 4 (STING high, miR-181a low). N Comparison of STING protein levels in low versus high miR-181a TNBC tumor tissues in the TMA (Student’s t-test). * p < 0.05, ** p < 0.01, *** p < 0.001. Cell viability assays were performed in triplicates

Article Snippet: Furthermore, a clinically annotated tissue microarray (TMA) for TNBC (#BR1301) was obtained from US Biomax (Derwood, MD).

Techniques: Over Expression, Quantitative RT-PCR, Plasmid Preparation, Western Blot, Comparison

Journal: Frontiers in Pharmacology

Article Title: Gallic acid suppresses the progression of triple-negative breast cancer HCC1806 cells via modulating PI3K/AKT/EGFR and MAPK signaling pathways

doi: 10.3389/fphar.2022.1049117

Figure Lengend Snippet: GA inhibited HCC1806 cells viability and affected cells morphology, and affected HCC1806 cells proliferation and cycle distribution. (A) The MDA-MB-468 cells viability was detected by MTT assay. (B) The HCC1806 cell inhibitory rate was detected by MTT assay. (C) The morphology changes of HCC1806 cells treated with different concentrations of GA were observed under an inverted microscope. (D,E) Effect of GA on the colony formation rate of HCC1806 cells. (F,G) The cell cycle distribution of HCC1806 were assessed by flow cytometry. Date were expressed as means ± SD. Compared with the control group, ** p < 0.01.

Article Snippet: The human TNBC HCC1806 and MDA-MB-468 cells line were purchased form Jiangsu KeyGEN BioTECH Corp., Ltd. (Nanjing, China).

Techniques: MTT Assay, Inverted Microscopy, Flow Cytometry, Control

Journal: Frontiers in Pharmacology

Article Title: Gallic acid suppresses the progression of triple-negative breast cancer HCC1806 cells via modulating PI3K/AKT/EGFR and MAPK signaling pathways

doi: 10.3389/fphar.2022.1049117

Figure Lengend Snippet: GA induced HCC1806 cells apoptosis accompanied by ROS accumulation and MMP depolarization. (A–C) Hoechst 33258 staining and flow cytometry were performed to assess the effect of GA on HCC1806 cells apoptosis. (D,E) The depolarization ratio of HCC1806 cells were detected by flow cytometry. (F,G) The ROS level of HCC1806 cells were assessed by a fluorescence microscope. Date were expressed as means ± SD. Compared with the control group, ** p < 0.01.

Article Snippet: The human TNBC HCC1806 and MDA-MB-468 cells line were purchased form Jiangsu KeyGEN BioTECH Corp., Ltd. (Nanjing, China).

Techniques: Staining, Flow Cytometry, Fluorescence, Microscopy, Control

Journal: Frontiers in Pharmacology

Article Title: Gallic acid suppresses the progression of triple-negative breast cancer HCC1806 cells via modulating PI3K/AKT/EGFR and MAPK signaling pathways

doi: 10.3389/fphar.2022.1049117

Figure Lengend Snippet: GA induces HCC1806 cells apoptosis via the mitochondrial pathway. (A–D) Effect of GA on the expression of Bax, Bcl-2, cleaved-Caspase-3, cleaved-Caspase-9 and P53 proteins in HCC1806 cells. (E) Effect of GA on the expression of Bax, Bcl-2, Caspase-3, Caspase-9 and P53 mRNA in HCC1806 cells. Date were expressed as means ± SD. Compared with the control group, * p < 0.05, ** p < 0.01.

Article Snippet: The human TNBC HCC1806 and MDA-MB-468 cells line were purchased form Jiangsu KeyGEN BioTECH Corp., Ltd. (Nanjing, China).

Techniques: Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Gallic acid suppresses the progression of triple-negative breast cancer HCC1806 cells via modulating PI3K/AKT/EGFR and MAPK signaling pathways

doi: 10.3389/fphar.2022.1049117

Figure Lengend Snippet: GA induces HCC1806 cells apoptosis via the PI3K/Akt/EGFR and MAPK signaling pathways. (A–D) Effect of GA on the expression of P-PI3K, P-AKT, P-EGFR, P-ERK1/2, P-JNK, and P-P38 proteins in HCC1806 cells. (E,F) Effect of GA on the expression of PI3K, AKT, EGFR, ERK1/2, JNK and P38 mRNA in HCC1806 cells. Date were expressed as means ± SD. Compared with the control group, * p < 0.05, ** p < 0.01.

Article Snippet: The human TNBC HCC1806 and MDA-MB-468 cells line were purchased form Jiangsu KeyGEN BioTECH Corp., Ltd. (Nanjing, China).

Techniques: Protein-Protein interactions, Expressing, Control

Journal: Molecular cancer therapeutics

Article Title: Nuclear Epidermal Growth Factor Receptor is a Functional Molecular Target in Triple-Negative Breast Cancer

doi: 10.1158/1535-7163.MCT-13-1021

Figure Lengend Snippet: (A) TNBC cells express total and activated forms of EGFR. Whole cell lysate was harvested from six TNBC cell lines and two HER2 positive cell lines. α-Tubulin was used as a loading control. (B) TNBC cells express nEGFR. Cell lines were harvested for nuclear proteins. α-Tubulin, calnexin, and Histone H3 were used as loading and purity controls, respectively. Confocal IF microscopy depicts nEGFR expression. Merged images were magnified to depict nEGFR (Confocal zoom, white arrows). A single Z-Slice image depicts overlap between blue and red signal (white dashed-line boxes). Magnification 600X. EGFR primary antibody specificity was validated with siEGFR and blocking peptides. (C) Immunogold labeling of nEGFR. TNBC cells were fixed and processed for transmission EM. CY, cytoplasm; NE, nuclear envelope; NUC, nucleus; NOS, nucleolus. Images were digitally zoomed to highlight gold particles in the nucleus (black arrows). (D) Human TNBC tumors express nEGFR. IHC staining for EGFR was performed on a total of 74 TNBC patient tumor sections. Representative cases demonstrating nEGFR expression are depicted (black arrows).

Article Snippet: Two human TNBC TMAs (#695711112B and #69572306) were purchased from TriStar Technology Group (Rockville, MD, USA).

Techniques: Control, Microscopy, Expressing, Blocking Assay, Labeling, Transmission Assay, Immunohistochemistry

Journal: Molecular cancer therapeutics

Article Title: Nuclear Epidermal Growth Factor Receptor is a Functional Molecular Target in Triple-Negative Breast Cancer

doi: 10.1158/1535-7163.MCT-13-1021

Figure Lengend Snippet: (A) Constitutively active Src (caSrc) enhances nEGFR translocation in TNBC cell lines. Cells were transfected with caSrc or an empty vector control for 48 hr prior to stimulation with EGF (5 nM, 45 min) to induce nEGFR translocation. Non-nuclear and nuclear proteins were harvested. nEGFR expression was quantitated using ImageJ software (B) A negative regulator of Src, src-like adaptor protein (SLAP), blocks nEGFR translocation in TNBC cell lines. Cells were transfected with SLAP-FLAG or an empty vector control for 48 hr prior to harvesting non-nuclear and nuclear proteins. nEGFR expression was analyzed. (C) SLAP can interact with EGFR and decrease EGFR activation at Tyrosine 1101. Cells were transfected with SLAP-FLAG or an empty vector control for 48 hr prior to harvesting whole cell lysate. 250 ug of cell lysate was immunoprecipitated with an anti-SLAP antibody. The same lysate was subjected to immunoblot analysis for activation of EGFR at Tyrosine 1101. pEGFR-Y1101 activity was quantitated using ImageJ software. Inset 1: EGFR mutated at Tyrosine 1101 is deficient in nuclear localization. Vector, EGFR-WT, and EGFR-Y1101F were transfected into CHOK1 cells for 48 hr prior to stimulation with EGF (5 nM, 45 min). Non-nuclear and nuclear proteins were harvested, and nEGFR expression was analyzed.

Article Snippet: Two human TNBC TMAs (#695711112B and #69572306) were purchased from TriStar Technology Group (Rockville, MD, USA).

Techniques: Translocation Assay, Transfection, Plasmid Preparation, Control, Expressing, Software, Activation Assay, Immunoprecipitation, Western Blot, Activity Assay

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Long Non-Coding RNA THOR Enhances the Stem Cell-Like Traits of Triple-Negative Breast Cancer Cells Through Activating β-Catenin Signaling

doi: 10.12659/MSM.923507

Figure Lengend Snippet: THOR directly binds to β-catenin mRNA, enhances its mRNA stability and expression. ( A–C ) The mRNA and protein levels of Sox9 and β-catenin were examined in TNBC cells with THOR knockdown or not. ( D ) The in vitro RNA-RNA interaction was performed to examine the THOR-β-catenin RNA interaction in TNBC cells. ( E ) RIP assay on THOR level in RNA pulled down by anti-β-catenin. ( F, G ) Analysis on the mRNA stability of β-catenin was performed in TNBC cells with THOR knockdown or not. ( H ) Luciferase reporter analysis on the TOP flash and FOP flash in MDA-MB-453 cells with THOR knockdown or not. ( I ) The expression of the downstream effectors of β-catenin was detected in TNBC cells with THOR knockdown or not. ** P <0.01. mRNA – messenger RNA; TNBC – triple negative breast cancer; RIP – RNA immunoprecipitation.

Article Snippet: Human TNBC lines MDA-MB-453 and MDA-MB-231, luminal B type of breast cancer cell line T-47D, luminal A type of breast cancer cell line MCF-7, and human mammary epithelial cell line MCF-10A cells were purchased from Sino Biological (Beijing, China).

Techniques: Expressing, In Vitro, Luciferase, Immunoprecipitation