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RNAseq experiment of non-exposed skin, Actinic Keratosis, Intraepdimeral Carcinoma and Squamous Cell Carcinoma

ABSTRACT: RNAseq experiment of non-exposed skin, Actinic Keratosis, Intraepdimeral Carcinoma and Squamous Cell Carcinoma

PROVIDER: PRJEB20787 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR1971040_1.fastq.gz	Fastqsanger.gz
	ERR1971040_2.fastq.gz	Fastqsanger.gz
	ERR1971050_1.fastq.gz	Fastqsanger.gz
	ERR1971050_2.fastq.gz	Fastqsanger.gz
	ERR1971060_1.fastq.gz	Fastqsanger.gz

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RNAseq experiment of non-exposed skin, actinic keratosis, intraepidermal carcinoma and squamous cell carcinoma

Project description:The goal of the study is to compare the transcriptomic profile of patient-derived skin samples, Actinic Keratosis, Intraepidermal Carcinoma and Squamous Cell Carcinoma samples using next generation RNA Sequencing. We aim to identify perturbed pathways and similarities between the 4 conditions. Please note that detailed patients' information, such as gender and age, was withheld from this submission to protect the patients' identity.

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Transcription profiling of human normal skin, actinic keratosis and squamous cell carcinoma

Project description:normal skin (no), actinic keratosis (ak), and squamous cell carcinoma (scc) of the skin were examined:; BACKGROUND: Carcinogenesis is a multi-step process indicated by several genes up- or down-regulated during tumor progression. This study examined and identified differentially expressed genes in cutaneous squamous cell carcinoma (SCC). RESULTS: Three different biopsies of 5 immunosuppressed organ-transplanted recipients each normal skin (all were pooled), actinic keratosis (AK) (two were pooled), and invasive SCC and additionally 5 normal skin tissues from immunocompetent patients were analyzed. Thus, total RNA of 15 specimens were used for hybridization with Affymetrix HG-U133A microarray technology containing 22,283 genes. Data analyses were performed by prediction analysis of microarrays using nearest shrunken centroids with the threshold 3.5 and ANOVA analysis was independently performed in order to identify differentially expressed genes (p < 0.05). Verification of 13 up- or down-regulated genes was performed by quantitative real-time reverse transcription (RT)-PCR and genes were additionally confirmed by sequencing. Broad coherent patterns in normal skin vs. AK and SCC were observed for 118 genes. CONCLUSION: The majority of identified differentially expressed genes in cutaneous SCC were previously not described.

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Progression from actinic keratosis to cutaneous squamous cell carcinoma involves miR-204 silencing and a switch towards STAT3 activation

Project description:Squamous cell carcinoma is the second most common skin cancer and frequently progress from an intraepithelial actinic keratosis. The role of microRNAs during the progression from actinic keratosis to cutaneous squamous cell carcinoma (cSCC) remains to be elicited. By using an Agilent microRNA expression microarray we found the expression of miR-204 to be markedly downregulated in cSCC when compared to actinic keratoses. DNA methylation of the TRPM3 promoter region upstream of miR-204-5p was identified as one of the repressive mechanisms that accounts for miR-204 silencing in cSCC. Functional studies on HaCaT cells revealed that this microRNA downregulates the Signal Transducer and Activator of Transcription 3 (STAT3) pathway and favours the MAPK signaling pathway, likely acting through PTPN11, a tyrosine phosphatase that is a direct miR-204 target. We found that activated STAT3, as detected by pY705-STAT3 immunofluorescence, is retained in the membrane and cytoplasm compartment in AK, whereas cSCC displayed STAT3 in the nuclei. Taken together, our data indicates that MiR-204 may act as a “rheostat” that controls the signaling towards the MAPK pathway or the STAT3 pathway.

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Project description:RNA-seq of human skin lesions diagnosed as actinic keratosis, intraepidermal carcinoma or squamous cell carcinoma

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Project description:Cutaneous squamous cell carcinoma (cSCC) is an important type of skin cancer. It is difficult to diagnose and treat, which seriously endanger people’s the physical and mental health. Actinic keratosis (AK) induced by ultraviolet radiation is the precancerous lesion of cSCC. The mechanism of occurrence and development of cSCC is still unclear, and the research on epigenetics is even less. In this project, we intend to study the mechanism of cSCC, especially the transformation from normal skin to AK, cSCC and the metastasis in late stage of cSCC, using multi-omics sequencing technology with experimental methods. Through the integrated analysis of multi-omics information such as DNA methylation, RNA methylation, chromatin accessibility and transcriptome, we obtained the epigenetic map of different stages of cSCC and identified the potential key epigenetic driving factors for cSCC. This project provided an important scientific basis for the clinical treatment and drug development of cSCC, as well as method reference for the study of other cancers.

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Single-cell Sequencing Highlights Heterogeneity and Malignant Progression in Actinic Keratosis and Cutaneous Squamous Cell Carcinoma

Project description:Cutaneous squamous cell carcinoma (cSCC) is the second most frequent of the keratinocyte-derived malignancies with actinic keratosis (AK) as a precancerous lesion. To comprehensively delineate the underlying mechanisms for the whole progression from normal skin to AK to invasive cSCC, we performed single-cell RNA-seq (scRNA-seq) to acquire the transcriptomes of 138,982 cells from 13 samples of six patients including AK, squamous cell carcinoma in situ (SCCIS), cSCC and their normal tissues, covering comprehensive clinical courses of cSCC. We identified diverse cell types, including important subtypes with different gene expression profiles and functions in major keratinocytes. In SCCIS, we discovered the malignant subtypes of basal cells with differential proliferative and migration potential. Differentially expressed genes (DEG) analysis screened out multiple key driver genes including transcription factors (TFs) along AK to cSCC progression. Immunohistochemistry (IHC) / immunofluorescence (IF) experiments and single-cell ATAC sequencing (scATAC-seq) data verified the expression changes of these genes. The functional experiments confirmed the important roles of these genes in regulating cell proliferation, apoptosis, migration and invasion in cSCC tumor. Furthermore, we comprehensively described the tumor microenvironment (TME) landscape and potential keratinocyte-TME crosstalk in cSCC providing theoretical basis for immunotherapy. Together, our findings provide a valuable resource for deciphering the progression from AK to cSCC and identifying potential targets for anticancer treatment of cSCC.

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