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PD-L1 ChIP-seq in MDA-MB-231 cells

ABSTRACT: PD-L1 ChIP-seq in MDA-MB-231 cells

PROVIDER: PRJNA611577 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11272700_1.fastq.gz	Fastqsanger.gz
	SRR11272700_2.fastq.gz	Fastqsanger.gz
	SRR11272701.fastq.gz	Fastqsanger.gz
	SRR11272702.fastq.gz	Fastqsanger.gz
	SRR11272699_1.fastq.gz	Fastqsanger.gz

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PD-L1 binding RNAs: MDA-MB-231

Project description:Analysis of PD-L1 binding RNAs using crosslinked RNA immunoprecipitation in MDA-MB-231.

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PD-L1 ChIP-seq in MDA-MB-231 cells

Project description:Although membrane-anchored PD-L1 has been well-studied for its engagement with PD-1 on T cells to evade anti-tumor immunity, whether PD-L1 regulate oncogenic signaling pathways in tumor cells remains elusive. We found that a portion of PD-L1 could translocate to the nucleus and knockout of PD-L1 changed RNA profiles in MDA-MB-231 cells. To further explore potential role of PD-L1 in regulating gene expression in tumor cells, we performed ChIP-seq in HA tag-inserted (after the signal peptide) PD-L1 re-expressed MBA-MB-231cells (endogenous PD-L1 knockout background). Methods: HA-insert-PD-L1 was re-introduced into MDA-MB-231 PD-L1 knockout cells using lentivirus and then the infected cells were selected with puromycin to make stable subclones. ChIP experiments were performed using HA antibody (Abcam, ab9110). Conclusions: Our study indicates that nuclear PD-L1 has potential roles in regulating gene transcription. More efforts are needed to further dissect the exact working model.

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Project description:Although membrane-anchored PD-L1 has been well-studied for its engagement with PD-1 on T cells to evade anti-tumor immunity, whether PD-L1 regulate oncogenic signaling pathways in tumor cells remains elusive. Methods: Total RNA from MDA-MB-231 wild type (WT) , MDA-MB-231 PD-L1 knock-out (KO), CT26 WT or CT26 Pd-l1 KO cells was purified using Qiagen RNeasy mini kit (Qiagen) according to the manufacturer’s instructions. Library preparation and sequenceing analysis were performed at the Molecular Genetics Core facility in Dana-Farber Cancer institute. RNA-seq data was aligned using STAR v2.5.4a. Conclusions: Our study indicates that PD-L1 may regulate genes that are involved in immune response regulating pathways.

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The miRNA profiling in MDA-MB-231, MDA-MB-231/E1A and knockdown of Dicer in MDA-MB-231/E1A cells

Project description:Dicer, RNase III endonuclease, is an essential enzyme in miRNA biogenesis that regulates target gene expression, and it has been reported that aberrant expressions of Dicer associate with the clinical outcomes of patients in various cancers. To explore the miRNA differencial expression regulated by Dicer in MDA-MB-231/E1A cells, the microarray profiling analysis was employed to conduct differentially expressed miRNAs in stable MDA-MB-231/vector, MDA-MB-231/E1A, and MDA-MB-231/E1A/shDicer cells.

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