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Expression analysis of KHR and KHR-NICD oral tumors

ABSTRACT: KHR-NICD oral tumors grow faster than KHR oral tumors. The hypothesis was to test for differences in gene expression between KHR and KHR-NICD oral tumors.

ORGANISM(S): Mus musculus

PROVIDER: GSE59950 | GEO | 2015/09/09

SECONDARY ACCESSION(S): PRJNA257156

REPOSITORIES: GEO

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Expression analysis of KHR and KHR-NICD oral tumors

Project description:KHR-NICD oral tumors grow faster than KHR oral tumors. The hypothesis was to test for differences in gene expression between KHR and KHR-NICD oral tumors. Total RNA was isolated for KHR (n=6) and KHR-NICD (n=6) oral tumors. Expression profiling was performed using the Illumina MouseRef-8 v2 (GPL6885) array. Studies were conducted using tumors from FVB x C57BL/6 F1 mice.

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Expression analysis of KHR and KR oral tumors with intact or deficient Notch1

Project description:KHR and KR tumors grow faster when deficient for Notch1. The hypothesis was to test for differences in gene expression between KHR and KHR oral tumors with intact or deficient Notch1 haplotypes. Total RNA was isolated for KHR (n=5), KHR-N+/- (n=4), KHR-N-/- (n=5), KR-N+/- (n=5), KR-N-/- (n=4) oral tumors. Expression profiling was performed using the Illumina MouseRef-8 v2 (GPL6885) array. Studies were conducted using tumors from C57BL/6 mice.

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Project description:Mutational landscape of Oral Squamous Cell Carcinoma (OSCC) is predominated by frequent inactivating mutations in EGF- like ligand binding domain of NOTCH1. However role of NOTCH1 signalling in tumorigenesis is highly context and cell type dependent. In this study, we investigated phenotypic effect of NOTCH1 mutations in primary line derived from OSCC biopsy. Based on gain-of-function assays in NOTCH1 mutant line we demonstrated regulation of cell behaviour and morphology, by NOTCH1 intracellular domain (NICD) leading to changes in cell proliferation, migration, clonal growth and differentiation. Importantly, overexpression of NICD results in upregulation of ETV7/TEL2 which negatively regulates SERPINE1 expression and confers malignant cells with less aggressive phenotype. Further knockdown of SERPINE1 expression simulated the phenotypes observed upon rescue of NICD. In accord with our cell-based model we observed strong correlation between NOTCH1, ETV7 and SERPINE1 expression in OSCC primary tumours indicating similar deregulated mechanisms during oral carcinogenesis. Overall this study suggests a tumour suppressive role of NOTCH1 in OSCC and highlights possible mechanisms leading to changes in cell behaviour and morphology in absence of NICD potentially by regulating TEL2 and SERPINE1 expression.

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Gene expression profiling of prostate tissues and seminal vesicles from PB-Pten-NICD mice

Project description:To investigate the identity of the tissue origin for metastases, we compared gene expression profiles of the prostate tissues and seminal vesicles from PB-Pten-NICD mice and those of 7 lung metastases from different mice. Three respective cell lines established from primary seminal vesicle tumors, primary prostate tumors, and lung metastases in PB-Pten-NICD mice were also included in the microarray analysis. We hypothesized that the gene expression profiles of primary and metastatic tumors should resemble those of their tissue origins. We identified genes differentially expressed between the seminal vesicle and prostate tumors. The expression profiles of the two cell lines established from primary tumors resembled those of their respective tissue origins, which supports our hypothesis. The expression profile of the metastatic cell line resembled those of the prostate tumor tissues, indicating the prostate as its tissue of origin. The expression profiles of 2 metastatic specimens each resembled those of the seminal vesicle tumors and the prostate tumors, respectively, while the expression profiles of the remaining 3 samples displayed a mixed pattern. This study suggests that metastatic tumors in this model may have originated from either the prostate, or the seminal vesicles, or both.

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Gene expression profile upon TNF-a stimulation, forced expression of NICD, or TNF-a stimulation under forced expression of NICD, in human colon carcinoma-derived LS174T cells

Project description:A human colon carcinoma-derived cell line LS174T was modified to overexpress NICD, intracellular domain of Notch1, upon doxycycline addition (designated as LS174T-tetON-NICD cells), using the T-rex system (Invitrogen). We have previously shown that these cells can overexpress NICD under the control of CMV promoter (Okamoto R et al, Am J Physiol, 296(1):G23-35, 2009), and the amont of the overexpressed NICD protein reaches to the maximal level in early as 3 hours from doxycycline addition (100ng/ml), which persists for up to 24 hours. In the present experiment, LS174T-tetON-NICD cells were treated with recombinant human TNF-a (50ng/ml) alone, doxycycline alone (100ng/ml), co-treated by both TNF-a and doxycycline, or left untreated (Control) for 24 hours, and subjected for analysis. Experiment was done using a modified sub-line of LS174T cell (LS174T-tetON-NICD cells), in which overexpression of NICD can be induced by a Doxycycline dependent manner.

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