Gene expression alteration by different cell lines derived from lung SCC of IKKM-NM-1 mutant mice
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ABSTRACT: In this study, we found that kinase-dead IKKM-NM-1 knockin (KAL) mice. develop spontaneous lung squamous cell carcinomas (SCCs) associated with IKKM-NM-1 downregulation and marked pulmonary inflammation.KKM-NM-1 downregulation dysregulates the expression of multiple oncogenes and tumor suppressors in K5+ lung epithelial cells. The mutant macrophages increase inflammatory responses and oxidative stress to promote DNA damage in IKKM-NM-1-mutant K5+ lung epithelial cells, which further dysregulate the levels of multiple oncogenes, tumor suppressors, and stem cell genes, thereby promoting the IKKM-NM-1lowK5+p63hi cell transition to tumor cells in L-IkkM-NM-1KA/KA lungs. To further investigate the underlying mechanisms by which the lung SCC development, we generated two cell lines, named as S1 and S2 individually, which were derived from KAL lung SCC. The S1 cells express high level of Sca1 and exhibit tumorigenic phenotype, while the S2 cells express low level of Sca1 and exhibit less tumorigenic phenotype. The aim of this microarray assay is to identify differentially expressed genes between S1 and S2 cells, which may highlight the important genes or pathways involved in inflammation-associated lung SCC carcinogenesis. There were two cell lines were used to perform Gene chip assay with Affymetrix. The two cell lines, named as S1 and S2 individually, were derived from KAL lung SCC. The S1 cells express high level of Sca1 and exhibit tumorigenic phenotype, while the S2 cells express low level of Sca1 and exhibit less tumorigenic phenotype.
Project description:In this study, we found that kinase-dead IKKα knockin (KAL) mice. develop spontaneous lung squamous cell carcinomas (SCCs) associated with IKKα downregulation and marked pulmonary inflammation.KKα downregulation dysregulates the expression of multiple oncogenes and tumor suppressors in K5+ lung epithelial cells. The mutant macrophages increase inflammatory responses and oxidative stress to promote DNA damage in IKKα-mutant K5+ lung epithelial cells, which further dysregulate the levels of multiple oncogenes, tumor suppressors, and stem cell genes, thereby promoting the IKKαlowK5+p63hi cell transition to tumor cells in L-IkkαKA/KA lungs. To further investigate the underlying mechanisms by which the lung SCC development, we generated two cell lines, named as S1 and S2 individually, which were derived from KAL lung SCC. The S1 cells express high level of Sca1 and exhibit tumorigenic phenotype, while the S2 cells express low level of Sca1 and exhibit less tumorigenic phenotype. The aim of this microarray assay is to identify differentially expressed genes between S1 and S2 cells, which may highlight the important genes or pathways involved in inflammation-associated lung SCC carcinogenesis.
Project description:Expression analysis of F1 haploid segregants from a cross between BY4716 (isogenic to S288c) and a wild isolate collected by R. Mortimer. Each segregant sample was subjected to a dye-swap pair of arrays. All arrays used the same pool of reference BY4716 sample. In sample titles, "BY" alone signifies the reference sample and all other strings represent segregants. All sample titles are of the form S1-S2. S1 indicates the sample whose signal was read as channel 1 (Cy5 labeling, fluorescence at 635 nm) and S2 as channel 2 (Cy3, 532 nm). Keywords: repeat sample
Project description:Expression analysis of BY4716(isogenic to S288c) and a wild isolate collected by R. Mortimer. Each strain was grown in culture 6 independent times and RNA from each culture was isolated. Each of these RNA samples was subjected to a dye-swap pair of arrays (except the "RM11" sample, which only got one array). All arrays used the same pool of reference BY4716 sample. In sample titles, "BY" alone signifies the reference sample and all other strings represent independent cultures. All sample titles are of the form S1-S2. S1 indicates the sample whose signal was read as channel 1 (Cy5 labeling, fluorescence at 635 nm) and S2 as channel 2 (Cy3, 532 nm). Keywords: repeat sample
Project description:Molecular Features of the Serological IgG Repertoire Elicited by Egg-based, Cell-based, or Recombinant HA Seasonal Influenza Vaccines. __sample information__ Donorname day0S1 or day28S2 Elu or FT Pf number A1 S1 FT 7649_JL_5a.raw A1 S1 FT 7649_JL_5b.raw A1 S1 FT 7649_JL_5c.raw A1 S2 FT 7649_JL_7a.raw A1 S2 FT 7649_JL_7b.raw A1 S2 FT 7649_JL_7c.raw A1 S1 Elu 7649_JL_6a.raw A1 S1 Elu 7649_JL_6b.raw A1 S1 Elu 7649_JL_6c.raw A1 S2 Elu 7649_JL_8a.raw A1 S2 Elu 7649_JL_8b.raw A1 S2 Elu 7649_JL_8c.raw A2 S1 FT 8078_JP_1a.raw A2 S1 FT 8078_JP_1b.raw A2 S1 FT 8078_JP_1c.raw A2 S2 FT 8078_JP_2a.raw A2 S2 FT 8078_JP_2b.raw A2 S2 FT 8078_JP_2c.raw A2 S1 Elu 8078_JP_3a.raw A2 S1 Elu 8078_JP_3b.raw A2 S1 Elu 8078_JP_3c.raw A2 S2 Elu 8078_JP_4a.raw A2 S2 Elu 8078_JP_4b.raw A2 S2 Elu 8078_JP_4c.raw A3 S1 FT 8116_JP_1a.raw A3 S1 FT 8116_JP_1b.raw A3 S1 FT 8116_JP_1c.raw A3 S2 FT 8116_JP_2a.raw A3 S2 FT 8116_JP_2b.raw A3 S2 FT 8116_JP_2c.raw A3 S1 Elu 8116_JP_3a.raw A3 S1 Elu 8116_JP_3b.raw A3 S1 Elu 8116_JP_3c.raw A3 S2 Elu 8116_JP_4a.raw A3 S2 Elu 8116_JP_4b.raw A3 S2 Elu 8116_JP_4c.raw A4 S1 FT 8149_JP_1a.raw A4 S1 FT 8149_JP_1b.raw A4 S1 FT 8149_JP_1c.raw A4 S2 FT 8149_JP_2a.raw A4 S2 FT 8149_JP_2b.raw A4 S2 FT 8149_JP_2c.raw A4 S1 Elu 8149_JP_3a.raw A4 S1 Elu 8149_JP_3b.raw A4 S1 Elu 8149_JP_3c.raw A4 S2 Elu 8149_JP_4a.raw A4 S2 Elu 8149_JP_4b.raw A4 S2 Elu 8149_JP_4c.raw A5 S1 FT 8209_JP_1a.raw A5 S1 FT 8209_JP_1b.raw A5 S1 FT 8209_JP_1c.raw A5 S2 FT 8209_JP_2a.raw A5 S2 FT 8209_JP_2b.raw A5 S2 FT 8209_JP_2c.raw A5 S1 Elu 8209_JP_3a.raw A5 S1 Elu 8209_JP_3b.raw A5 S1 Elu 8209_JP_3c.raw A5 S2 Elu 8209_JP_4a.raw A5 S2 Elu 8209_JP_4b.raw A5 S2 Elu 8209_JP_4c.raw B1 S1 FT 7924_JL_1a.raw B1 S1 FT 7924_JL_1b.raw B1 S1 FT 7924_JL_1c.raw B1 S2 FT 7924_JL_2a.raw B1 S2 FT 7924_JL_2b.raw B1 S2 FT 7924_JL_2c.raw B1 S1 Elu 7924_JL_4a.raw B1 S1 Elu 7924_JL_4b.raw B1 S1 Elu 7924_JL_4c.raw B1 S2 Elu 7924_JL_5a.raw B1 S2 Elu 7924_JL_5b.raw B1 S2 Elu 7924_JL_5c.raw B2 S1 FT 7947_JL_1a.raw B2 S1 FT 7947_JL_1b.raw B2 S1 FT 7947_JL_1c.raw B2 S2 FT 7947_JL_2a.raw B2 S2 FT 7947_JL_2b.raw B2 S2 FT 7947_JL_2c.raw B2 S1 Elu 7947_JL_3a.raw B2 S1 Elu 7947_JL_3b.raw B2 S1 Elu 7947_JL_3c.raw B2 S2 Elu 7947_JL_4a.raw B2 S2 Elu 7947_JL_4b.raw B2 S2 Elu 7947_JL_4c.raw B3 S1 FT 8129_JL_1a.raw B3 S1 FT 8129_JL_1b.raw B3 S1 FT 8129_JL_1c.raw B3 S2 FT 8129_JL_2a.raw B3 S2 FT 8129_JL_2b.raw B3 S2 FT 8129_JL_2c.raw B3 S1 Elu 8129_JL_3a.raw B3 S1 Elu 8129_JL_3b.raw B3 S1 Elu 8129_JL_3c.raw B3 S2 Elu 8129_JL_4a.raw B3 S2 Elu 8129_JL_4b.raw B3 S2 Elu 8129_JL_4c.raw B4 S1 FT 8164_JP_1a.raw B4 S1 FT 8164_JP_1b.raw B4 S1 FT 8164_JP_1c.raw B4 S2 FT 8164_JP_2a.raw B4 S2 FT 8164_JP_2b.raw B4 S2 FT 8164_JP_2c.raw B4 S1 Elu 8164_JP_3a.raw B4 S1 Elu 8164_JP_3b.raw B4 S1 Elu 8164_JP_3c.raw B4 S2 Elu 8164_JP_4a.raw B4 S2 Elu 8164_JP_4b.raw B4 S2 Elu 8164_JP_4c.raw B5 S1 FT 8237_JP_1a.raw B5 S1 FT 8237_JP_1b.raw B5 S1 FT 8237_JP_1c.raw B5 S2 FT 8237_JP_2a.raw B5 S2 FT 8237_JP_2b.raw B5 S2 FT 8237_JP_2c.raw B5 S1 Elu 8237_JP_3a.raw B5 S1 Elu 8237_JP_3b.raw B5 S1 Elu 8237_JP_3c.raw B5 S2 Elu 8237_JP_4a.raw B5 S2 Elu 8237_JP_4b.raw B5 S2 Elu 8237_JP_4c.raw C1 S1 FT 8108_JP_1a.raw C1 S1 FT 8108_JP_1b.raw C1 S1 FT 8108_JP_1c.raw C1 S2 FT 8108_JP_2a.raw C1 S2 FT 8108_JP_2b.raw C1 S2 FT 8108_JP_2c.raw C1 S1 Elu 8108_JP_3a.raw C1 S1 Elu 8108_JP_3b.raw C1 S1 Elu 8108_JP_3c.raw C1 S2 Elu 8108_JP_4a.raw C1 S2 Elu 8108_JP_4b.raw C1 S2 Elu 8108_JP_4c.raw C2 S1 FT 8048_JL_1a.raw C2 S1 FT 8048_JL_1b.raw C2 S1 FT 8048_JL_1c.raw C2 S2 FT 8048_JL_2a.raw C2 S2 FT 8048_JL_2b.raw C2 S2 FT 8048_JL_2c.raw C2 S1 Elu 8048_JL_3a.raw C2 S1 Elu 8048_JL_3b.raw C2 S1 Elu 8048_JL_3c.raw C2 S2 Elu 8048_JL_4a.raw C2 S2 Elu 8048_JL_4b.raw C2 S2 Elu 8048_JL_4c.raw C3 S1 FT 8068_JP_1a.raw C3 S1 FT 8068_JP_1b.raw C3 S1 FT 8068_JP_1c.raw C3 S2 FT 8068_JP_2a.raw C3 S2 FT 8068_JP_2b.raw C3 S2 FT 8068_JP_2c.raw C3 S1 Elu 8068_JP_3a.raw C3 S1 Elu 8068_JP_3b.raw C3 S1 Elu 8068_JP_3c.raw C3 S2 Elu 8108_JP_5a.raw C3 S2 Elu 8108_JP_5b.raw C3 S2 Elu 8108_JP_5c.raw C4 S1 FT 8203_JP_1a.raw C4 S1 FT 8203_JP_1b.raw C4 S1 FT 8203_JP_1c.raw C4 S2 FT 8203_JP_2a.raw C4 S2 FT 8203_JP_2b.raw C4 S2 FT 8203_JP_2c.raw C4 S1 Elu 8203_JP_3a.raw C4 S1 Elu 8203_JP_3b.raw C4 S1 Elu 8203_JP_3c.raw C4 S2 Elu 8203_JP_4a.raw C4 S2 Elu 8203_JP_4b.raw C4 S2 Elu 8203_JP_4c.raw C5 S1 FT 8258_JP_1a.raw C5 S1 FT 8258_JP_1b.raw C5 S1 FT 8258_JP_1c.raw C5 S2 FT 8258_JP_2a.raw C5 S2 FT 8258_JP_2b.raw C5 S2 FT 8258_JP_2c.raw C5 S1 Elu 8258_JP_3a.raw C5 S1 Elu 8258_JP_3b.raw C5 S1 Elu 8258_JP_3c.raw C5 S2 Elu 8258_JP_4a.raw C5 S2 Elu 8258_JP_4b.raw C5 S2 Elu 8258_JP_4c.raw
Project description:Objectives: The contribution of circular RNAs (circRNAs) in the regulation of embryonic lung development is not yet clear. This study aims to uncover the differentially expressed circRNAs in mouse embryonic lung tissue. Methods: High-throughput sequencing (HTS) was used to determine the cirRNA expression profiles at four time points of mouse embryonic lung tissue [embryonic (E) Day 14.5 (E14.5/S1), (E16.5/S2), (E18.5/S3), and newborn (N) Day 7.5 (N7.5/S4)]. CircRNAs that were significantly differential were subjected to bioinformatic analysis and functional prediction for their host genes. Finally, quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) was conducted to examine their levels. Results: Dynamically expressed circRNAs in the mouse embryonic lung tissue were identified. After that, a total of 231 circRNAs differentially expressed between the three groups (S1 vs. S2, S1 vs. S3 and S1 vs. S4) in embryonic lung tissue (fold change ≥ 2.0; p < 0.05). These circRNAs were subjected to bioinformatic analysis to predict their potential biological functions. Finally, six circRNAs (circRNA3_Ttn, circRNA56_Galnt18, circRNA38_Alg12, circRNA42_Filip1l, circRNA43_Ptprm, and circRNA57_Ncoa3) that were further validated using qRT-PCR. Conclusions: These findings illustrate the vital functions of circRNAs in mouse embryonic lung development. Combined with our results and previous researches, circRNAs may serve as potential miRNA sponges in the embryonic lung development. Our research is conducive to further explore the development of embryonic lungs, and the dynamic expressed circRNAs may be novel molecules that regulate mouse lung development.
Project description:Lung squamous cell carcinoma (SCC) is a deadly disease for which current treatments are inadequate. We demonstrate that bi-allelic inactivation of Lkb1 and Pten in the mouse lung led to SCC that recapitulated the histology, gene expression and microenvironment found in human disease. Lkb1/Pten-null (LP) tumors expressed the squamous markers Krt5, p63 and Sox2, and transcriptionally resembled the basal subtype of human SCC. In contrast to mouse adenocarcinomas, the LP tumors contained immune populations enriched for tumor-associated neutrophils. Sca1+/Ngfr+ fractions were enriched for tumor propagating cells (TPCs) that could serially transplant the disease in orthotopic assays. TPCs in the LP model and Ngfr+ cells in human SCCs highly expressed Pdl1, suggesting a novel mechanism of immune escape for TPCs. We used microarrays to detail the gene expression profles among lung SCC tumor epitheial cell, lung ADC tumor epithelia cell and normal epithelial cells.
Project description:Lung squamous cell carcinoma (SCC) is a deadly disease for which current treatments are inadequate. We demonstrate that bi-allelic inactivation of Lkb1 and Pten in the mouse lung led to SCC that recapitulated the histology, gene expression and microenvironment found in human disease. Lkb1/Pten-null (LP) tumors expressed the squamous markers Krt5, p63 and Sox2, and transcriptionally resembled the basal subtype of human SCC. In contrast to mouse adenocarcinomas, the LP tumors contained immune populations enriched for tumor-associated neutrophils. Sca1+/Ngfr+ fractions were enriched for tumor propagating cells (TPCs) that could serially transplant the disease in orthotopic assays. TPCs in the LP model and Ngfr+ cells in human SCCs highly expressed Pdl1, suggesting a novel mechanism of immune escape for TPCs. We used microarrays to detail the gene expression profles among lung SCC tumor epitheial cell, lung ADC tumor epithelia cell and normal epithelial cells.
Project description:Lung squamous cell carcinoma (SCC) is a deadly disease for which current treatments are inadequate. We demonstrate that bi-allelic inactivation of Lkb1 and Pten in the mouse lung led to SCC that recapitulated the histology, gene expression and microenvironment found in human disease. Lkb1/Pten-null (LP) tumors expressed the squamous markers Krt5, p63 and Sox2, and transcriptionally resembled the basal subtype of human SCC. In contrast to mouse adenocarcinomas, the LP tumors contained immune populations enriched for tumor-associated neutrophils. Sca1+/Ngfr+ fractions were enriched for tumor propagating cells (TPCs) that could serially transplant the disease in orthotopic assays. TPCs in the LP model and Ngfr+ cells in human SCCs highly expressed Pdl1, suggesting a novel mechanism of immune escape for TPCs. We used microarrays to detail the gene expression profles among lung SCC tumor epitheial cell, lung ADC tumor epithelia cell and normal epithelial cells. Kras tumor stroma cells and LP tumor stroma cells were sorted by FACS, the cells were gated as EpCAM-/CD45+/CD31+
Project description:Lung squamous cell carcinoma (SCC) is a deadly disease for which current treatments are inadequate. We demonstrate that bi-allelic inactivation of Lkb1 and Pten in the mouse lung led to SCC that recapitulated the histology, gene expression and microenvironment found in human disease. Lkb1/Pten-null (LP) tumors expressed the squamous markers Krt5, p63 and Sox2, and transcriptionally resembled the basal subtype of human SCC. In contrast to mouse adenocarcinomas, the LP tumors contained immune populations enriched for tumor-associated neutrophils. Sca1+/Ngfr+ fractions were enriched for tumor propagating cells (TPCs) that could serially transplant the disease in orthotopic assays. TPCs in the LP model and Ngfr+ cells in human SCCs highly expressed Pdl1, suggesting a novel mechanism of immune escape for TPCs. We used microarrays to detail the gene expression profles among lung SCC tumor epitheial cell, lung ADC tumor epithelia cell and normal epithelial cells. Normal EpCAM+, Kras tumor EpCAM+ and LP tumor EpCAM+ were sorted by FACS, the cells were gating as EpCAM+/CD45-/CD31-
Project description:We have reported that pulmonary infiltrating macrophages, which highly express inducible nitric-oxide synthase (iNOS/NOS2), play a critical role for driving spontaneous lung SCC development in a new mouse model (L-Ikka KA/KA; KA/KA) that mimics human lung SCC development. However, the role of NOS2 in lung SCC development is unknown. Our data suggests deletion of the gene iNOS prevents tumorogenesis in this mouse model. We use microarray data to compare levels of expression of genes associated with lung tumor development and prevention.