Activity of lapatinib is independent of EGFR expression level in HER2-overexpressing breast cancer cells

Abstract Background : MiR-218-5p is a small non-coding RNA acting as either oncogenes or tumor suppressor genes in human cancer. The expression levels of some miRNAs in human breast cancer plays a potential role in disease pathogenesis. Methods : Thirty pairs of invasive ductal carcinoma and adjacent specimens were included in the study. Breast tissues cell lines MCF-7 and MDA-MB-231 were identified as a breast cancer research cell line. MiR-218-5p mimics, miR-218-5p inhibitor, or negative controls were transfected. Specific antibodies were probed with LRIG1, ErbB2, and EGFR. Proliferation, migration, cell cycle and apoptosis, dual-luciferase reporter assay and immunohistochemistry were used to analyze miR-218-5p、LRIG1 and so on. Results : It was shown that miR-218-5p expression was higher in 30 breast cancer specimens than adjacent normal breast tissues. In human breast cancer cells MCF-7 and MDA-MB-231, restoring miR-218-5p promoted cell proliferation and migration and inhibited cell apoptosis and cell cycle arrest in the G1 stage. Luciferase assays indicated miR-218-5p could bind with its putative target site in the 3'-untranslated region (3'-UTR) of LRIG1. RT-qPCR, western blot, and immunocytochemistry analyses all indicated miR-218-5p overexpression results in LRIG1 downregulation at the mRNA and protein levels. ErbB2 and EGFR were found to be downstream effectors of miR-218-5p. Conclusion : MiR-218-5p promotes ErbB2 and EGFR expression by inhibiting LRIG1 in breast cancer cells, which suggests miR-218-5p and LRIG1 may act as an oncogene in breast cancer and it could be used as a therapeutic target for breast cancer treatments. Keywords: Breast cancer; miR-218-5p; LRIG1; Oncogene

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Expression of genes modulated by epigallocatechin-3-gallate in breast cancer cells

Herba Polonica ◽

10.2478/hepo-2018-0016 ◽

2018 ◽

Vol 64 (3) ◽

pp. 31-37

Author(s):

Anna Bogacz ◽

Marlena Wolek ◽

Bogna Juskowiak ◽

Monika Karasiewicz ◽

Adam Kamiński ◽

...

Keyword(s):

Breast Cancer ◽

Adjuvant Therapy ◽

Mrna Expression ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Molecular Mechanisms ◽

Mrna Level ◽

Expression Level ◽

Mrna Levels ◽

Mrna Expression Level

Summary Introduction: Breast cancer is the most common malignant cancer among women. Both drug resistance and metastasis are major problems in the treatment of breast cancer. Therefore, adjuvant therapy may improve patients’ survival and affect their quality of life. It is suggested that epigallocatechin gallate (EGCG) which is well known for its chemopreventive activity and acts on numerous molecular targets may inhibit the growth and metastasis of some cancers. Hence, discovering the metastatic molecular mechanisms for breast cancer may be useful for therapy. Objective: The aim of the study was to determine the effect of EGGC on the mRNA expression level of genes such as ZEB1, ABCB1, MDM2, TWIST1 and PTEN in MCF-7 breast cancer cells. Methods: MCF7/DOX were cultured in the presence of 0.2 μM DOX and EGCG (20-50 μM). The mRNA expression level was determined by real-time quantitative PCR using RealTime ready Custom Panel 96 kit. Results: Our results showed an important increase (about 2-fold for 20 μM EGCG + 0.2 μM DOX and 2.5-fold for 50 μM EGCG + 0.2 μM DOX, p<0.05) in ZEB1 expression levels. In case of ABCB1 gene lack of influence on the mRNA level was observed (p>0.05). We also observed significant decrease of ZEB1 expression in MCF7 cells with 20 μM and 50 μM EGCG (p<0.05). In addition, EGCG (20 μM) caused an increase of MDM2 and PTEN mRNA levels in almost 100% (p<0.05) and 40% (p>0.05), respectively. Lack of the influence of EGCG was noted for the TWIST1 gene expression. In case of MCF7/DOX we showed an increase of mRNA level of PTEN gene about 50% (p<0.05). Conclusions: These results suggest that EGCG may be potentially used in adjuvant therapy in the breast cancer treatment.

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Anti-proliferative and apoptotic effect of tetrahydrobenzo[h]quinoline on MCF-7 human breast cancer cell

Pharmaceutical Sciences ◽

10.34172/ps.2021.58 ◽

2021 ◽

Author(s):

Maryam Ghaffari ◽

Dariush Shanehbandi ◽

Solmaz Sarhadi ◽

Mina Hanifeh Ahagh ◽

Mahsa Maleki Moghaddam ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Expression Level ◽

Caspase 8 ◽

Human Breast ◽

Caspase 9 ◽

Anti Cancer ◽

Apoptotic Effect ◽

Mcf 7

Background: Quinoline and its derivatives display various biological activities based on versatility in designing a new drug class for medicinal applications. Hence, synthesizing innovative and varied derivatives of quinoline has gained considerable attention among chemists and biologists. This study evaluated the anti-proliferative and apoptotic effect of tetrahydrobenzo[h]quinoline on Michigan Cancer Foundation-7 (MCF-7) human breast cancer cells. Methods: The anti-proliferative effect of tetrahydrobenzo[h]quinoline was studied via MTT [3 0-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] assays. A quantitative and qualitative study of apoptosis was carried out via flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Quantitative real-time PCR (qPCR) and immunoblotting analysis were employed to identify the expression level of genes and proteins involved in the apoptosis signaling pathway. Results: The synthesized compound reduced 50% of cell growth at concentrations of 10 and 7.5 µM during 24 and 48h, respectively, and induced apoptosis up to 30% in MCF-7 cancer cells. Regarding the gene expression level, Bcl-2 displayed considerable alleviation, whereas Bax expression increased significantly. Despite the remarkable increase in caspase 9 expression, there was no noticeable difference in the caspase 8 expression in treated cells compared to the control group. Western blotting data showed that the protein expression level of Bcl-2, pro-caspase 8, and 9 reduced. The protein content of Bax, cleaved-caspase 8, and 9 increased significantly, of which the protein level of cleaved-caspase 9 exhibited a tremendous rise in the treated group. Conclusion: The newly synthesized tetrahydrobenzo[h]quinoline can be a promising organic compound for cancer treatment if its anti-cancer effect investigates by other types of breast cancer cells. In vivo studies should be used to investigate the anti-cancer efficiency of this compound.

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FRA-1 Expression Level Modulates Regulation of Activator Protein-1 Activity by Estradiol in Breast Cancer Cells

Molecular Endocrinology ◽

10.1210/mend.12.7.0133 ◽

1998 ◽

Vol 12 (7) ◽

pp. 973-985 ◽

Cited By ~ 34

Author(s):

Alexandre Philips ◽

Catherine Teyssier ◽

Florence Galtier ◽

Corinne Rivier-Covas ◽

Jean-Marc Rey ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Expression Level ◽

Activator Protein 1 ◽

Activator Protein

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EGFR Expression in HER2-Driven Breast Cancer Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21239008 ◽

2020 ◽

Vol 21 (23) ◽

pp. 9008

Author(s):

Florian Weinberg ◽

Diana B. Peckys ◽

Niels de Jonge

Keyword(s):

Breast Cancer ◽

Electron Microscopy ◽

Plasma Membrane ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Simultaneous Detection ◽

Orphan Receptor ◽

Membrane Expression ◽

Intact Cells ◽

Egfr Expression

The epidermal growth factor receptor HER2 is overexpressed in 20% of breast cancer cases. HER2 is an orphan receptor that is activated ligand-independently by homodimerization. In addition, HER2 is able to heterodimerize with EGFR, HER3, and HER4. Heterodimerization has been proposed as a mechanism of resistance to therapy for HER2 overexpressing breast cancer. Here, a method is presented for the simultaneous detection of individual EGFR and HER2 receptors in the plasma membrane of breast cancer cells via specific labeling with quantum dot nanoparticles (QDs). Correlative fluorescence microscopy and liquid phase electron microscopy were used to analyze the plasma membrane expression levels of both receptors in individual intact cells. Fluorescent single-cell analysis of SKBR3 breast cancer cells dual-labeled for EGFR and HER2 revealed a heterogeneous expression for receptors within both the cell population as well as within individual cells. Subsequent electron microscopy of individual cells allowed the determination of individual receptors label distributions. QD-labeled EGFR was observed with a surface density of (0.5–5) × 101 QDs/µm2, whereas labeled HER2 expression was higher ranging from (2–10) × 102 QDs/µm2. Although most SKBR3 cells expressed low levels of EGFR, an enrichment was observed at large plasma membrane protrusions, and amongst a newly discovered cellular subpopulation termed EGFR-enriched cells.

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Selective Growth Inhibition of Human Breast Cancer Cells by Graviola Fruit Extract In Vitro and In Vivo Involving Downregulation of EGFR Expression

Nutrition and Cancer ◽

10.1080/01635581.2011.563027 ◽

2011 ◽

Vol 63 (5) ◽

pp. 795-801 ◽

Cited By ~ 45

Author(s):

Yumin Dai ◽

Shelly Hogan ◽

Eva M. Schmelz ◽

Young H. Ju ◽

Corene Canning ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Human Breast Cancer ◽

Breast Cancer Cells ◽

Human Breast Cancer Cells ◽

Human Breast ◽

Fruit Extract ◽

Egfr Expression

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Tim-3 promotes cell aggressiveness and paclitaxel resistance through the NF-κB /STAT3 signaling pathway in breast cancer

10.21203/rs.2.22315/v1 ◽

2020 ◽

Author(s):

Yizi Cong ◽

Yuxin Cui ◽

Shiguang Zhu ◽

Jianqiao Cao ◽

Guangdong Qiao ◽

...

Keyword(s):

Breast Cancer ◽

Gene Expression ◽

Tight Junction ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Prognostic Significance ◽

Gene Expression Level ◽

Expression Level ◽

Cancer Tissue ◽

Junction Molecules

Abstract Background: T-cell immunoglobulin and mucin-domain containing molecule-3 (Tim-3) has been recognized as a promising target for cancer immunotherapy, but its exact role in breast cancer has not been fully elucidated. Methods: The gene expression level of Tim-3 in breast cancer and its prognostic significance were analysed. In vitro functions and associated mechanisms were then explored through establishing Tim-3 overexpressing breast cancer cells. Results: The gene expression level of Tim-3 was significantly higher (p<0.001) in breast cancer tissue compared to normal tissue following pooled analysis of the TCGA database. Tim-3 was a prognosis indicator in breast cancer patients as shown by KM-plotter (RFS: p=0.004; OS: p=0.099). Overexpression of the Tim-3 in Tim-3 low breast cancer cells promoted aggressiveness of breast cancer cells including proliferation, migration, invasion, tight junction deterioration and tumour-associated tubal formation. Furthermore, Tim-3 enhanced cellular resistance to paclitaxel. Tim-3 exerted its function by activating the NF-κB/STAT3 signalling pathway, and mediating gene regulation (upregulating CCND1, C-Myc, MMP1, TWIST, VEGF while downregulating E-cadherin). Additionally, Tim-3 downregulated tight junction molecules: ZO-2, ZO-1 and Occludin, which might further facilitate the tumour progression. Conclusions: Tim-3 plays a tumour-promoting role in breast cancer, suggesting that targeting Tim-3 may acquire a clinical benefit in antitumor therapy. Keywords: breast cancer, Tim-3, tight junction, chemoresistance, aggressiveness.

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