Preferential Killing of Breast Tumor Initiating Cells by <i>N,N</i>-Diethyl-2-[4-(Phenylmethyl)Phenoxy]Ethanamine/Tesmilifene

Deng, Tao; Liu, Jeff C.; Pritchard, Kathleen I.; Eisen, Andrea; Zacksenhaus, Eldad

doi:10.1158/1078-0432.ccr-08-1708

Cited by 19 publications

(7 citation statements)

References 52 publications

(33 reference statements)

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“…In our study, culturing tumor cells from unsorted pleural effusions under non-adherent culture conditions was successful in 77.8% of the patient samples. This success rate is similar to other studies, where 42–73% of spheroid cultures from pleural effusion aspirates could be initiated [32–34]. However, after sorting for putative breast cancer stem cell markers, the spheroid formation efficiency in these subpopulations was diminished.…”

Section: Discussionsupporting

confidence: 88%

Genetic profiling of putative breast cancer stem cells from malignant pleural effusions

et al. 2017

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A common symptom during late stage breast cancer disease is pleural effusion, which is related to poor prognosis. Malignant cells can be detected in pleural effusions indicating metastatic spread from the primary tumor site. Pleural effusions have been shown to be a useful source for studying metastasis and for isolating cells with putative cancer stem cell (CSC) properties. For the present study, pleural effusion aspirates from 17 metastatic breast cancer patients were processed to propagate CSCs in vitro. Patient-derived aspirates were cultured under sphere forming conditions and isolated primary cultures were further sorted for cancer stem cell subpopulations ALDH1+ and CD44+CD24-/low. Additionally, sphere forming efficiency of CSC and non-CSC subpopulations was determined. In order to genetically characterize the different tumor subpopulations, DNA was isolated from pleural effusions before and after cell sorting, and compared with corresponding DNA copy number profiles from primary tumors or bone metastasis using low-coverage whole genome sequencing (SCNA-seq). In general, unsorted cells had a higher potential to form spheres when compared to CSC subpopulations. In most cases, cell sorting did not yield sufficient cells for copy number analysis. A total of five from nine analyzed unsorted pleura samples (55%) showed aberrant copy number profiles similar to the respective primary tumor. However, most sorted subpopulations showed a balanced profile indicating an insufficient amount of tumor cells and low sensitivity of the sequencing method. Finally, we were able to establish a long term cell culture from one pleural effusion sample, which was characterized in detail. In conclusion, we confirm that pleural effusions are a suitable source for enrichment of putative CSC. However, sequencing based molecular characterization is impeded due to insufficient sensitivity along with a high number of normal contaminating cells, which are masking genetic alterations of rare cancer (stem) cells.

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Section: Discussionsupporting

confidence: 88%

Genetic profiling of putative breast cancer stem cells from malignant pleural effusions

et al. 2017

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“…Of note, 0.5-to 1-cm mammary tumors were minced and digested with 100 U/mL collagenase/hyaluronidase (STEMCELL Technologies) for 60 minutes at 37 C. Following negative selection with the EasySep Kit (STEMCELL Technologies), enriched Lin À mammary epithelial cells were plated onto ultra-low attachment 24-well plates (Corning, Costar) in Dulbecco's Modified Eagle Medium (DMEM)/F-12 HAM medium containing basic fibroblast growth factor (bFGF), EGF, and B-27 supplement as described previously (9,20) Mouse short hairpin RNA kinase library (TRC1) and Tbk1 shRNA clones A subset of the Broad Institute RNAi Consortium (TRC) pLKO.1 short hairpin RNA (shRNA) library targeting 520 mouse kinase genes with 2,567 lentiviral clones (4-5 shRNAs/gene) were used for screens in a total of 28-Â 96-well plates (21). Each plate included 3 to 5 wells with empty vector (Lenti-GFP) as negative controls.…”

Section: Methodsmentioning

confidence: 99%

“…On day 3, medium was gently aspirated and replaced with 200 mL fresh medium. On day 7, tumorsphere counts and MTT assays were carried out (20).…”

Section: Lentiviral Shrna Screensmentioning

confidence: 99%

shRNA Kinome Screen Identifies TBK1 as a Therapeutic Target for HER2+ Breast Cancer

et al. 2014

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“…Cell viability and tumor sphere formation were measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, as previously described [16]. …”

Section: Methodsmentioning

confidence: 99%

PTEN Loss-Mediated Akt Activation Increases the Properties of Cancer Stem-Like Cell Populations in Prostate Cancer

Kim

Bae

Hong³

et al. 2014

Oncology

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Objective: To demonstrate that the PTEN/PI3K/Akt/NF-κB pathway plays an important role in regulating the prostate cancer stem-like cell population by upregulating ABCG2. Methods: Targeted PTEN knockdown in human prostate DU145 and 22Rv1 cells using a small interfering RNA were confirmed by immunoblot analysis using antibodies of PTEN, phospho-Akt, Akt, and a-tubulin. Knockdown PTEN DU145 and 22Rv1 cells were augmented, and the stem cell-like properties were examined by cell viability and tumor sphere formation and treated by Akt IV inhibitor to provide the signal transduction pathway. Luciferase activity assays were performed. Results: The knockdown of PTEN in prostate cancer cell lines increased the stem-like properties of the cells, including their sphere-forming ability, stem cell population number, epithelial-mesenchymal transition-related gene expression, and ABCG2 expression. Additionally, PTEN expression was highly associated with elevated expression of phospho-Akt. Treatment with an Akt inhibitor suppressed the PTEN-mediated effects on the properties of these stem-like cells as well as drug resistance, ABCG2 expression, and the NF-κB pathway. Conclusion: The loss of PTEN in prostate cancer cells resulted in an increased PI3K/Akt pathway. Due to the Akt activation, PTEN loss may play an important role in prostate cancer by promoting cancer stemness through a mechanism that involves enhanced NF-κB signaling.

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Preferential Killing of Breast Tumor Initiating Cells by N,N-Diethyl-2-[4-(Phenylmethyl)Phenoxy]Ethanamine/Tesmilifene

Cited by 19 publications

References 52 publications

Genetic profiling of putative breast cancer stem cells from malignant pleural effusions

Genetic profiling of putative breast cancer stem cells from malignant pleural effusions

shRNA Kinome Screen Identifies TBK1 as a Therapeutic Target for HER2+ Breast Cancer

PTEN Loss-Mediated Akt Activation Increases the Properties of Cancer Stem-Like Cell Populations in Prostate Cancer

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