Human breast cancer MCF-7 cells: SN38-selected cells
Ontology highlight
ABSTRACT: Transcriptional profiling of human breast cancer MCF-7 cells comparing control untreated cells with cells exposed to step-wise increasing doses of SN38, an active metabolite of irinotecan.
Project description:Transcriptional profiling of human breast cancer MCF-7 cells comparing control untreated cells with cells exposed to step-wise increasing doses of SN38, an active metabolite of irinotecan. MCF-7 cell clones were established by a limiting dilution method. SN38-sensitive MCF-7 clone (Clone 3) was used for further analyses. MCF-7/SNR cells were established by continuously exposing this clone to SN38 with stepwise increasing concentrations ranging from 0.5 ng/mL to 50 ng/mL over a period of six months. MCF-7/SNR cells were maintained in the presence of 50 ng/mL SN38 until 24 h prior to any experiment. Total RNA was extracted from MCF-7 and MCF-7/SNR cells for cDNA microarray analysis.
Project description:Human breast cancer cell line MCF-7 is usually sensitive to chemotherapy drug BMS-554417, an insulin receptor (IR) and insulin-like growth factor receptor (IGFR) inhibitor. However, through step-wise increase in BMS-554417 doses in culture media, we were able able to screen and select a single MCF-7 clone that is BMS-554417 resistant. It is cross resistant to BMS-536924. This new line of MCF-7 cells was named as MCF-7R4. The transcriptome profiling of both MCF-7 and MCF-7R4 was performed using Affymetrix HG-U133 plus2.0 GeneChip arrays.
Project description:The effects of several compounds on the MCF7 human adenocarcinoma mammary cell line were analysed by gene expression profiling. Tested compounds: HSP90 inhibitors: 17AAG (Tanespimycin), NVP-AUY922, NMS-E973 (cpd developed at NMS). CDK inhibitors: CDK-887 (cpd developed at NMS). Topoisomerase inhibitors: Doxorubicin, SN38 (active metabolite of Irinotecan). The MCF7 cell line was treated with the different compounds for 6 hours at a dose equal to 5 times the IC50. Untreated MCF7 cells were used as a control. Two replicates per treatment.
Project description:Human breast cancer cell line MCF-7 is usually sensitive to chemotherapy drug BMS-554417, an insulin receptor (IR) and insulin-like growth factor receptor (IGFR) inhibitor. However, through step-wise increase in BMS-554417 doses in culture media, we were able able to screen and select a single MCF-7 clone that is BMS-554417 resistant. It is cross resistant to BMS-536924. This new line of MCF-7 cells was named as MCF-7R4. The transcriptome profiling of both MCF-7 and MCF-7R4 was performed using Affymetrix HG-U133 plus2.0 GeneChip arrays. Five replicates of MCF-7 and five replicates of MCF-7R4 were profiled.
Project description:Tumors consist of heterogeneous cell population, containing cancer cell subpopulations with anticancer drug-resistant property, called “persister” cells. To reveal the character of the persister cells, we analyzed gene expression profile of patient-derived gastric cells and residual cancer cells after treatment with 5-FU or SN38, an active metabolite of irinotecan. In our study, we identified ALDH1A3 as a marker and a cell proliferation factor of persister cells. To examine molecular pathways regulated by ALDH1A3, we analyzed gene expression profile of patient-derived gastric JSC15-3 in which ALDH1A3 was knocked down by using shRNAs.
Project description:HCT116 p53 null cells were treated with 5nM SN38 (active metabolite of CPT-11) for 6, 12 and 24h. Following this RNA was harvested for transcriptional profiling.
Project description:Using 4 primary melanoma cell lines for which autologous tumor-infiltrating T lymphocytes (TILs) were available, a screen of clinically-relevant small-molecule inhibitors (SMIs) was performed to find SMIs that could synergistically enhance tumor cell killing by autologous TILs. Among positive results were SMIs targeting topoisomerase I (TOP1) or HSP90. Of note, as implied by the fact that these SMIs had synergistic effects, there was relatively little direct cytotoxicity of the SMIs when used alone. Gene expression profiling was undertaken to identify changes induced by SMIs of TOP1 or HSP90. SN38 is the active metabolite of irinotecan, a standard-of-care clinical TOP1 inhibitor. Ganetespib is a newer generation HSP90 inhibitor, reported to exhibit greater potency in preclinical tumor models and reduced toxicity in rodents, compared to other 1st and 2nd generation HSP90 inhibitors, consistent with its favorable safety profile in patients.
Project description:We have performed sucrose-gradient-based isolation of polysomal fractions from untreated and TGF-beta treated MCF-10A and MCF7 cells, subjected these fractions to RNA-seq, and also sequenced total mRNA from each cell line in the treated and untreated condition
Project description:Radiotherapy is a commonly used treatment modality for the local control of breast cancer. However, the clinical signs of radiotherapy response are often not apparent for several weeks post-treatment. We currently lack tools to predict and/or monitor tumor responses during treatment. The aim of this study was to identify tumor secreted biomarkers of radiotherapy response. Estrogen receptor positive (ER+) MCF-7 cells were serum-starved for 2 h, and were then exposed to various doses of radiation (0, 2, 4, 6, 8 or 10 Gy). Conditioned media (CM) was harvested at 1, 2, 4, 8 and 24 h post-radiation for each radiation dose. Samples underwent processing for liquid chromatography-mass spectrometry (LC-MS) following collection. 33 proteins at the 24 h time point were found to have significantly increased secretion levels (up to 12-fold) at all radiation doses compared to untreated cells. To validate the secretomic results and to further investigate the potential use of these proteins as biomarkers of radiosensitivity, the secreted protein levels of candidate biomarkers were assessed through western blot (WB). WB analysis was performed using CM samples to assess the secretion levels from parental and radioresistant (RR) cell lines (developed within our lab) 24 h after the cells had received a single radiation dose of 2 Gy. Secretion levels of our candidate biomarkers were found to be significantly increased from the radiosensitive MCF-7 cells treated with 2 Gy of radiation at 24 h compared to 24 h untreated controls. In comparison, candidate biomarker levels in the CM samples from untreated and radiation-treated MCF-7 RR cells remained low. Currently there are no validated predictive biomarkers to monitor radiotherapy responses during treatment. These biomarkers could have a clinical role in personalising radiotherapy dosing schedules and durations for solid tumours in the neoadjuvant and palliative setting.