{"database":"biostudies-arrayexpress","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown","Transcriptomics","Genomics","Proteomics"],"submitter":["Domenico Memoli"],"instrument_platform":["Illumina NovaSeq 6000"],"study_type":["RNA-seq of coding RNA"],"organism":["Homo sapiens"],"species":["Homo sapiens"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/E-MTAB-15334"],"description":["Background: cancer therapy is one of the challenging arguments in constant development. One of the principal problems connected with the tumor therapy arise form the potential side effects connected with the classical chemotherapeutic treatment but also with the molecular target therapy. The identification of novel molecules useful for the reduction of potential side effects but also as a new therapeutic opportunity is one of the hot topics. (2) Methods: we identified a Castalin from the chestnut shells from the NRM. We treated different cancer cell lines with Castalin alone or in combination with CHK1 inhibitor. Finally, we performed a RNaseq analysis of HeLa cells treated with Castalin. (3) Results: we demonstrated the ability of Castalin to induce DNA damage, probably by inducing an in-crease of ROS production. Consistently, the antioxidant treatment, with Ascorbic acid, re-duced the DNA damage induced with Castalin. Finally, we demonstrated the potential synergistic effect of Castalin with SRA737, a CHK1 inhibitor currently used in clinical trial (4) Conclusions: we demonstrated the ability of Castalin to induce DNA damage favoring a NHEJ repair. Moreover, the use of Castalin in combination with SRA737 increase the ef-ficacy of CHK1 inhibitor reducing its possible side effects."],"repository":["biostudies-arrayexpress"],"sample_protocol":["Sample Collection - Cervix adenocarcinoma cell line (HeLa), human triple-negative breast cancer (TNBC) model cell line (MDA-MB-231) and pleural effusion of hormone-dependent human breast carcinoma (MCF-7) were obtained the American Type Culture Collection (ATCC). HeLa and MBA-MD 231 were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Thermo Fisher Scientific, Waltham, MA, USA). MCF7 cell line was cultured in Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA). All media were supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific), 100 U/mL penicillin, 100 µg/mL streptomycin, and 2 mM L-glutamine. Cells were maintained at 37 °C in a humidified incubator with 5% CO₂.","Library Construction - The RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina using manufacturer’s instructions (New England Biolabs, Ipswich, MA, USA). Briefly, mRNAs were initially enriched with Oligod(T) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized. cDNA fragments were end repaired and adenylated at 3’ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by PCR with limited cycles. The sequencing library was validated on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by using Qubit 3.0 Fluorometer (ThermoFisher Scientific, Waltham, MA, USA) as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA).","Nucleic Acid Extraction - Total RNA was extracted from frozen cell samples using Qiagen RNeasy Plus Universal mini kit following manufacturer’s instructions (Qiagen, Hilden, Germany).     RNA samples were quantified using Qubit 3.0 Fluorometer (ThermoFisher Scientific, Waltham, MA, USA) and RNA integrity was checked with 4200 TapeStation (Agilent Technologies, Palo Alto, CA, USA).","Sequencing - The sequencing libraries were multiplexed and clustered onto a flowcell on the Illumina NovaSeq instrument according to manufacturer’s instructions. The samples were sequenced using a 2x150bp Paired End (PE) configuration. Image analysis and base calling were conducted by the NovaSeq Control Software (NCS). Raw sequence data (.bcl files) generated from Illumina NovaSeq was converted into fastq files and de-multiplexed using Illumina bcl2fastq 2.20 software. One mis-match was allowed for index sequence identification"],"figure_sub":["Organization","MINSEQE Score","Assays and Data","MAGE-TAB Files"],"pubmed_authors":["Domenico Memoli"],"additional_accession":[]},"is_claimable":false,"name":"Castalin induces ROS production, leading to DNA damage and increasing the activity of CHK1 inhibitor in cancer cell lines","description":"Background: cancer therapy is one of the challenging arguments in constant development. One of the principal problems connected with the tumor therapy arise form the potential side effects connected with the classical chemotherapeutic treatment but also with the molecular target therapy. The identification of novel molecules useful for the reduction of potential side effects but also as a new therapeutic opportunity is one of the hot topics. (2) Methods: we identified a Castalin from the chestnut shells from the NRM. We treated different cancer cell lines with Castalin alone or in combination with CHK1 inhibitor. Finally, we performed a RNaseq analysis of HeLa cells treated with Castalin. (3) Results: we demonstrated the ability of Castalin to induce DNA damage, probably by inducing an in-crease of ROS production. Consistently, the antioxidant treatment, with Ascorbic acid, re-duced the DNA damage induced with Castalin. Finally, we demonstrated the potential synergistic effect of Castalin with SRA737, a CHK1 inhibitor currently used in clinical trial (4) Conclusions: we demonstrated the ability of Castalin to induce DNA damage favoring a NHEJ repair. Moreover, the use of Castalin in combination with SRA737 increase the ef-ficacy of CHK1 inhibitor reducing its possible side effects.","dates":{"release":"2025-09-04T00:00:00Z","modification":"2025-09-04T10:35:00.666Z","creation":"2025-07-22T17:25:09.311Z"},"accession":"E-MTAB-15334","cross_references":{"ENA":["ERP177069"],"EFO":["EFO_0002944","EFO_0004170","EFO_0005518","EFO_0003738","EFO_0004184"]}}