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Differential gene expression after ingenol mebutate (IM) treatment

ABSTRACT: We treated primary keratinocytes and SCC cells with IM for 24h to identify differentially expressed genes after treatment compared to DMSO contorl

ORGANISM(S): Homo sapiens

PROVIDER: GSE63308 | GEO | 2015/05/03

SECONDARY ACCESSION(S): PRJNA267521

REPOSITORIES: GEO

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Differential gene expression after ingenol mebutate (IM) treatment

Project description:We treated primary keratinocytes and SCC cells with IM for 24h to identify differentially expressed genes after treatment compared to DMSO contorl 3x DMSO keratinocytes, 3x IM 100 nM IM keratinocytes, 3x 10 uM IM keratinocytes, 3x DMSO SCC cells, 3x 1 nM IM SCC cells, 3x 100 nM IM SCC cells, 3x 10 uM IM SCC cells

2015-05-03 | E-GEOD-63308 | biostudies-arrayexpress

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Project description:BACKGROUND & AIMS: TLR4 is an innate immune receptor with expression in human skin, keratinocytes as well as squamous cell carcinoma (SCC) of the skin. In the present study we investigate the role of TLR4 as a negative regulator of keratinocyte proliferation. We present here that the expression of TLR4 increased with the differentiation of cultured keratinocytes in a passage-dependent manner or under calcium-rich conditions. Moreover, the down-regulation of TLR4 by specific knockdown increased the proliferation of primary normal, SCC-derived and HaCaT keratinocytes in vitro. In addition, subcutaneously injected HaCaT keratinocytes with shTLR4 formed growing tumors in nude mice. In contrast, we observed lower proliferation and increased migration in vitro of the SCC13 cell line stably overexpressing TLR4 in comparison to SCC13 TLR4 negative cells. In vivo, SCC13 TLR4-overexpressing tumors showed delayed growth in comparison to TLR4 negative tumors. The overexpression of TLR4 in SCC13 tumor cells was followed by phosphorylation of ERK1/2 and JNK and increased expression of ATF3. In gene expression arrays, the overexpression of TLR4 in tumor cells correlated with gene expression of ATF-3, IL-6, CDH13, CXCL-1, and TFPI. In summary, TLR4 negatively regulates the proliferation of primary keratinocytes and its overexpression reduces tumor growth of SCC cells.

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Skin cancer-associated S. aureus downregulates DNA repair mechanisms, and promotes oxidative stress and DNA damage in human keratinocytes

Project description:Actinic keratoses (AK) are premalignant lesions common on photo-damaged skin that, over time, can progress to squamous cell carcinoma (SCC). A high bacterial load of Staphylococcus aureus is associated with AK and SCC but it is unknown whether this has a direct impact on skin cancer development. To determine whether S. aureus is able to trigger pro-tumorigenic skin responses, we performed RNAseq and shotgun proteomics on primary human keratinocytes after challenge with sterile culture supernatant (‘secretome’) from S. aureus clinical strains isolated from AK and SCC. Certain S. aureus secretomes induced keratinocytes to overexpress SCC biomarkers that have been associated with skin carcinogenesis, and upregulate the expression of enzymes linked with reduced skin barrier function. Further, S. aureus secretomes downregulated DNA repair mechanisms and induced oxidative stress markers. Subsequent experiments confirmed that exposure to SCC-derived S. aureus secretomes lead to increased intracellular ROS levels and DNA damage in primary human keratinocytes. Altogether, this study reveal a novel mechanism for the pro-tumorigenic activity of S. aureus. Further studies are required to determine whether S. aureus products promote SCC development in vivo, which would have important implications for the treatment of AK and prevention of SCC.

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