Project description:Carbonic anhydrase IX (CA 9) is a transmembrane isoform of carbonic anhydrase (CA) that contributes to an acidification of tumor microenvironment. The expression of CA 9 in cervical tumors was shown to be strongly involved in high incidence of metastasis and poor prognosis. To search for the key regulators of metastasis related to ectopic expression of CA 9, we investigated differentially expressed gene profiles in CA 9- transfected cervix carcinoma cell line C33-A (CF) compared with mock-transfected (CP) cell line, using Affymetrix Human Genome U133 Plus 2.0 Array.
Project description:In order to explore the functions of carbonic anhydrase VI (CAVI) more fully, we examined the transcriptomic responses to CAVI deficiency in the submandibular gland, stomach, and duodenum of Car6-/- mice by cDNA microarray. 94, 56, and 127 genes were up- or down-regulated in the above-mentioned tissues of Car6-/- mice, respectively. The functional clustering of differentially expressed genes revealed a number of altered biological processes. In the duodenum, the significantly affected biological pathways included immune system process and retinol metabolic process. Response to oxidative stress and brown fat cell differentiation changed remarkably in the submandibular gland. Notably, the submandibular gland, stomach, and duodenum shared one prominent transcriptional susceptibility pathway-catabolic process. Submandibular gland, stomach, and duodenum samples were collected from three wild-type and three Car6-/- female mice, respectively, at the age of two months. Total RNAs were purified and used for cDNA microarray.
Project description:In order to explore the functions of carbonic anhydrase VI (CAVI) more fully, we examined the transcriptomic responses to CAVI deficiency in the submandibular gland, stomach, and duodenum of Car6-/- mice by cDNA microarray. 94, 56, and 127 genes were up- or down-regulated in the above-mentioned tissues of Car6-/- mice, respectively. The functional clustering of differentially expressed genes revealed a number of altered biological processes. In the duodenum, the significantly affected biological pathways included immune system process and retinol metabolic process. Response to oxidative stress and brown fat cell differentiation changed remarkably in the submandibular gland. Notably, the submandibular gland, stomach, and duodenum shared one prominent transcriptional susceptibility pathway-catabolic process.
Project description:Carbonic anhydrase IX (CA 9) is a transmembrane isoform of carbonic anhydrase (CA) that contributes to an acidification of tumor microenvironment. The expression of CA 9 in cervical tumors was shown to be strongly involved in high incidence of metastasis and poor prognosis. To search for the key regulators of metastasis related to ectopic expression of CA 9, we investigated differentially expressed gene profiles in CA 9- transfected cervix carcinoma cell line C33-A (CF) compared with mock-transfected (CP) cell line, using Affymetrix Human Genome U133 Plus 2.0 Array. CF and CP stable cell-lines trasfected respectively with full-length human CA 9 cDNA cloned into the vector pcDNA3.0 and empty vector control were used for RNA extraction and hybridization on affymetrix microarrays.
Project description:Background Carbonic anhydrases (CAs) are a family of enzymes that regulate pH homeostasis in various tissues. CA IX is an exceptional member of this family because in addition to the basic CA function, it has been implicated in several other physiological and pathological processes. Functions suggested for CA IX include roles in cell adhesion and malignant cell invasion. In addition, CA IX likely regulates cell proliferation and differentiation, which was demonstrated in Car9‾/‾ mice. These mice had gastric pit cell hyperplasia and depletion of chief cells; however, the specific molecular mechanisms behind the observed phenotypes remain unknown. Therefore, we wanted to study the effect of CA IX deficiency on whole-genome gene expression in gastric mucosa. This was done using Illumina Sentrix Mouse-6 Expression BeadChip arrays. The expression of several genes with notable fold-change values was confirmed by QRT-PCR. Results CA IX deficiency caused the induction of 86 genes and repression of 46 genes in the gastric mucosa. There was 92.9% concordance between the results obtained by microarray analysis and QRT-PCR. The differentially expressed genes included those involved in developmental processes and cell differentiation. In addition, CA IX deficiency altered the expression of genes responsible for immune responses and downregulated the expression of several digestive enzymes. Conclusions Microarray analysis identified several potential genes whose altered expression could explain the disturbed cell lineage phenotype in the Car9‾/‾ gastric mucosa. The results also indicated a novel role for CA IX in the regulation of immunologic processes and digestion. These findings reinforce the concept that the main role of CA IX is not the regulation of pH in the stomach mucosa. Instead, it is needed for proper function of several physiological processes.
Project description:In Nannochloropsis oceanica IMET1, transcript knockdown of a cytosolic carbonic anhydrase (CA2; g2018) specifically inhibited by HC resulted in ~45%, ~30% and ~40% elevation of photosynthetic oxygen evolution rate, growth rate and biomass accumulation rate under high CO2 (5% ), respectively. This CA2-knockdown mutant is demonated as M2. To probe mechanistic links underlying the mutant (M2; RNAi-knockdown line of carbonic anhydrase (CA2)) phenotypes, temporal transcriptomic profiles are compared between RNAi-knockdown line of carbonic anhydrase (CA2) and WT, at 12 h and 24h under high CO2 (5%).
Project description:Hypoxia is an important clinicopathological feature of solid tumours that limits response to therapy. Carbonic Anhydrase IX (CAIX) is a hypoxia-induced, pH regulator that has emerged as a viable therapeutic target. Targeting CAIX has demonstrated promise in difficult to treat preclinical models of breast pancreatic, melanoma and brain cancers when combined with chemotherapy and immunotherapy, however resistance ultimately develops and this mechanism is unclear. Here we used an unbiased genome-wide synthetic lethal CRISPR knock-out screen in basal breast cancer cells to identify co-vulnerabilities and mechanisms of compensation for CA9 loss. We identified a redox homeostasis network containing thioredoxin and the iron-sulfur cluster enzyme, cysteine desulfurase, NFS1, indicative of the critical importance of CAIX in this role.
Project description:cea05-01_carbonic-anhydrase - carbonic anhydrase (1 or 2) simple or double mutants - Identification of genes differentially expressed in leaves of single CA1 and CA2 T-DNA insertional mutants and in the corresponding double mutant vs wild type - CA1 (At3g01500) and CA2 (At5g14740) T-DNA insertional mutant lines, the double (CA1+CA2) mutant and wild type Arabidopsis seeds were sown in soil in a phytotron. Leaves were harvested 40 days later for RNA extraction
Project description:cea05-01_carbonic-anhydrase - carbonic anhydrase (1 or 2) simple or double mutants - Identification of genes differentially expressed in leaves of single CA1 and CA2 T-DNA insertional mutants and in the corresponding double mutant vs wild type - CA1 (At3g01500) and CA2 (At5g14740) T-DNA insertional mutant lines, the double (CA1+CA2) mutant and wild type Arabidopsis seeds were sown in soil in a phytotron. Leaves were harvested 40 days later for RNA extraction 6 dye-swap - gene knock out
Project description:In order to explore the functions of carbonic anhydrase VI (CA VI) more fully, we examined the transcriptomic responses to CA VI deficiency in the trachea, and lung of Car6-/- mice by cDNA microarray. 44 and 2 genes were up- or down-regulated in the above-mentioned tissues of Car6-/- mice, respectively. The functional clustering of differentially expressed genes revealed a number of altered biological processes. In the trachea, the affected biological pathways included metabolic process, biological regulation, single-organism process, and immune response in mucosal-associated lymphoid tissue. Trachea, and lung samples were collected from eight wild-type and six Car6-/- male mice, respectively, at the age of two months. Total RNAs were purified and used for cDNA microarray.