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RNA-Seq expression profiling of clonal populations from the OCI-C5x patient-derived ovarian clear cell carcinoma cell line

ABSTRACT: RNA-Seq expression profiling of clonal populations from the OCI-C5x patient-derived ovarian clear cell carcinoma cell line

PROVIDER: PRJNA508736 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8285540_1.fastq.gz	Fastqsanger.gz
	SRR8285540_2.fastq.gz	Fastqsanger.gz
	SRR8285541_1.fastq.gz	Fastqsanger.gz
	SRR8285541_2.fastq.gz	Fastqsanger.gz
	SRR8285542_1.fastq.gz	Fastqsanger.gz

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RNA-Seq expression profiling of clonal populations from the OCI-C5x patient-derived ovarian clear cell carcinoma cell line

Project description:Intratumoral heterogeneity is well-recognized but its impact on tumor progression is unclear. Here, we derived unbiased clonal populations from OCI-C5x, a patient-derived ovarian clear cell carcinoma cell line, and assessed their tumorigenicity and tracked their fate during tumor progression. Results provide insight into intratumoral heterogeneity of clear cell carcinoma transcriptomes.

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H3K4me2 profiling in OCI-Ly1 and OCI-Ly7 cell lines.

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H3K4me2 profiling in OCI-Ly1 and OCI-Ly7 cell lines.

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Gene expression in clonogenic versus non-clonogenic in pooled early passage OCI/AML-4 single cells

Project description:Identification of genes that regulate clonogenicity of AML cells is hindered by the difficulty of isolating pure populations of cells with defined proliferative abilities. Using early passages of the OCI/AML-4 cell line we sought to determine genes differentially expressed between clonogenic and non-clonogenic cells. We previously shown that OCI/AML-4 clonal siblings display coordinated growth patterns, so that the behaviour of a single cell that is destroyed in the generation of global single cell cDNA may be predicted by the growth patterns of its clonal siblings. Cells were plated in 96 well plates at limiting dilutions. Localized clusters of four cells were identified and micromanipulated such that three of the constituent cells were placed separately into individual microtitre wells containing growth medium and a feeder layer of OP9 cells, while the fourth cell was lysed and processed for global RT-PCR. The cells in each culture well were counted at 2 -3 day intervals until growth stopped. In this manner cDNA was generated from individual clonogenic OCI/AML-4 cells (sibling cells able to generate between 9-100 cells) or from individual non-clonogenic OCI/AML-4 cells (sibling cells were not able to generate more than 8 cells). Respective single cell cDNA was then pooled and compared by hybridization to cDNA microarrays. Using early passages of the OCI/AML-4 cell line cDNA was obtained from 17 clonogenic single cells (sibling cells able to generate between 9-100 cells) and 20 non-clonogenic single cells (sibling cells were not able to generate more than 8 cells). These cDNAs were pooled and hybridized to the cDNA microarray was conducted as three replicates, including one replicate which was a dye-swap.

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Clonal lineage from normal endometrium to ovarian clear cell carcinoma through ovarian endometriosis.

Project description:Clear cell carcinoma of the ovary is thought to arise from endometriosis. In addition, retrograde menstruation of shed endometrium is considered the origin of endometriosis. However, little evidence supports cellular continuity from uterine endometrium to clear cell carcinoma through endometriosis at the genomic level. Here, we performed multiregional whole-exome sequencing to clarify clonal relationships among uterine endometrium, ovarian endometriosis and ovarian clear cell carcinoma in a 56-year-old patient. Many somatic mutations including cancer-associated gene mutations in ARID1A, ATM, CDH4, NRAS and PIK3CA were shared among epithelium samples from uterine endometrium, endometriotic lesions distant from and adjacent to the carcinoma, and the carcinoma. The mutant allele frequencies of shared mutations increased from uterine endometrium to distant endometriosis, adjacent endometriosis, and carcinoma. Although a splice site mutation of ARID1A was shared among the four epithelium samples, a frameshift insertion in ARID1A was shared by adjacent endometriosis and carcinoma samples, suggesting that the biallelic mutations triggered malignant transformation. Somatic copy number alterations, including loss of heterozygosity events at PIK3CA and ATM, were identified only in adjacent endometriosis and carcinoma, suggesting that mutant allele-specific imbalance is another key factor driving malignant transformation. By reconstructing a clonal evolution tree based on the somatic mutations, we showed that the epithelium samples were derived from a single ancestral clone. Although the study was limited to a single patient, the results from this illustrative case could suggest the possibility that epithelial cells of ovarian endometriosis and clear cell carcinoma were descendants of uterine endometrial epithelium.

| S-EPMC7419022 | biostudies-literature

Gene expression in clonogenic versus non-clonogenic in pooled early passage OCI/AML-4 single cells

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Project description:Nucleostemin (NS; product of the GNL3 gene) is a nucleolar/nucleoplasm shuttling GTPase whose levels are high in stem cells while rapidly decreasing upon differentiation. NS levels are also high in several solid and hematological neoplasms, including acute myeloid leukemia (AML). While a role in telomere maintenance, response to stress stimuli and in favoring DNA repair has been proposed in solid cancers, little or no information is available as to the role of nucleostemin in AML. Here we investigate this issue with a proteomics approach. We use as a model system the OCI-AML 3 cell line harboring a heterozygous mutation at the NPM1 gene, which is the most frequent driver mutation in AML (approximately 30% of total AML cases). We show that NS is highly expressed in this cell line but, contrary to what previously shown in other cancers, its presence is dispensable for cell growth and viability. However, proteomics analysis of OCI-AML 3 cell line before and after nu-cleostemin (NS) silencing showed several effects in different biological functions as highlighted by ingenuity pathway analysis (IPA). In particular, we report an effect in down-regulating DNA repair through homologous recombination and we confirmed higher DNA damage rate in OCI-AML 3 cells when NS is depleted, which considerably increases upon stress induced by the topoisomerase II inhibitor etoposide.

2022-08-11 | PXD034012 | Pride

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