Isolated Small Rat Hepatocytes Express both Annexin III and Terminal Differentiated Hepatocyte Markers, Tyrosine Aminotransferase and Tryptophan Oxygenase, at the mRNA Level

Niimi, Shingo; Hyuga, Masashi; Harashima, Mizuho; Seki, Taiichiro; Ariga, Toshio; Kawanishi, Toru; Hayakawa, Tetsuo

doi:10.1248/bpb.27.1864

Cited by 5 publications

(1 citation statement)

References 23 publications

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“…15) Recently, we found that AnxA3 protein and its mRNA are not expressed in isolated parenchymal rat hepatocytes. 16,17) Consistent with these findings, AnxA3 protein and its mRNA are not detectable by Western blot analysis and Northern blot analysis in rat liver. [18][19][20][21] However, there have been no reports on the behavior of AnxA3 in primary cultured parenchymal rat hepatocytes.…”

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confidence: 54%

Expression of Annexin A3 in Primary Cultured Parenchymal Rat Hepatocytes and Inhibition of DNA Synthesis by Suppression of Annexin A3 Expression Using RNA Interference

Niimi

Harashima

Gamou

et al. 2005

Biological & Pharmaceutical Bulletin

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Annexin A3 is a member of the lipocortin/annexin family, which binds to phospholipids and membranes in a Ca 2؉ -dependent manner. Although annexin A3 has various functions in vitro, its cellular significance is completely unknown. Annexin A3 is not found in rat liver in vivo. In the present study, we investigated the expression of annexin A3 in primary cultured parenchymal rat hepatocytes. Annexin A3 protein was detected in 48-h, but not 2.5-h, cultured hepatocytes using Western blot analysis. The annexin A3 level further increased after an additional 24 h of culture. Annexin A3 mRNA was not detected in 2.5-h cultured hepatocytes but was detected 22 h after the start of culture by RT-PCR analysis, reaching a maximum value after 48 h of culture. To define the role of Annexin A3 in DNA synthesis, RNA interference was used to reduce annexin III gene expression in hepatocytes. The transfection of small interfering RNAs targeting annexin A3 in the hepatocytes reduced the corresponding mRNA and protein expression by approximately 80% and more than 90%, respectively, at 24 h after transfection. In the annexin A3 small interfering RNAs-transfected cells, DNA synthesis, as assessed by [ 3 H]thymidine incorporation, decreased by approximately 70% not only in the control cultures, but also in the hepatocyte growth factor-or epidermal growth factor-treated cells. These findings show that annexin A3 is expressed in primary cultured parenchymal rat hepatocytes and that the suppression of annexin A3 expression using RNA interference inhibits DNA synthesis.

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Section: ϩsupporting

confidence: 54%

Expression of Annexin A3 in Primary Cultured Parenchymal Rat Hepatocytes and Inhibition of DNA Synthesis by Suppression of Annexin A3 Expression Using RNA Interference

Niimi

Harashima

Gamou

et al. 2005

Biological & Pharmaceutical Bulletin

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Proteome alteration of early‐stage differentiation of mouse embryonic stem cells into hepatocyte‐like cells

Kang

Guo

et al. 2009

Electrophoresis

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To explore the molecular basis of inducible differentiation of embryonic stem cells into hepatocyte-like cells, a proteomic strategy was utilized to examine the global protein expression alterations after early-stage differentiation of a mouse D3 embryonic stem (ES) cell line along hepatic lineage. The undifferentiated D3 cells were treated stepwise with combinations of defined chemicals and growth factors. The differentiated cells were identified by hepatocyte-like morphology, expressed liver-specific markers as well as the evidence of glycogen storage. The subsequent proteomic separation and identification were performed with 2-DE followed by MALDI-TOF-MS/MS analysis. Of the 119 differentially displayed protein spots analyzed, 90 spots presenting 64 distinct proteins were finally identified. The interested protein expressions were validated by Western blotting such as albumin and cytokeratin-8. Bioinformatic annotations indicated that this set of proteins was enriched with transcription, translation regulation and protein processing, energy/metabolism and chaperone functions. A part of them had been found to be involved in the differentiation of mouse ES cells. Interestingly, approximately 40% of these proteins had been previously reported as being dysregulated in hepatocellular carcinoma. It suggested that these changed proteins may be candidate regulators of ES cell differentiation, some of them may be specific to hepatic differentiation.

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Differential radioactive quantification of protein abundance ratios between benign and malignant prostate tissues: Cancer association of annexin A3

et al. 2007

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A differential quantitative protein expression study, comparing matched prostate cancerous and benign tissues from 31 patients, revealed proteins newly associated with prostate cancer. Average effects for 17 proteins whose abundance was significantly different (p<0.01) across patients ranged from 1.5- to 6.1-fold, and included a number of known cancer markers. The most differentially abundant proteins between cancer and benign samples were isopeptidase T, serum amyloid P (SAP), annexin A3 (ANXA3) and mitochondrial enoyl coenzyme-A hydratase. SAP is restricted to stroma in healthy tissue, and the lower abundance in tumours may be explained by the reduced stromal content. ANXA3 is present in healthy epithelial cells, exhibits strong staining in precancerous prostatic intraepithelial neoplasia, and is relatively less abundant in individual tumour cells of increasing Gleason pattern (GP), despite exhibiting higher overall tissue abundance in tumours. ANXA3 staining was predominantly cytoplasmic, yet nuclear localization was also observed. Strongly staining single cells, possibly phagocytes, were interspersed in highly dedifferentiated GP5 tumour areas among tumour cells without measurable ANXA3. Local recurrent androgen ablation therapy-resistant tumours exhibit heterogenous low levels of ANXA3 staining. Results are discussed focussing on the potential implications for tumour tissues.

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Isolated Small Rat Hepatocytes Express both Annexin III and Terminal Differentiated Hepatocyte Markers, Tyrosine Aminotransferase and Tryptophan Oxygenase, at the mRNA Level

Cited by 5 publications

References 23 publications

Expression of Annexin A3 in Primary Cultured Parenchymal Rat Hepatocytes and Inhibition of DNA Synthesis by Suppression of Annexin A3 Expression Using RNA Interference

Expression of Annexin A3 in Primary Cultured Parenchymal Rat Hepatocytes and Inhibition of DNA Synthesis by Suppression of Annexin A3 Expression Using RNA Interference

Proteome alteration of early‐stage differentiation of mouse embryonic stem cells into hepatocyte‐like cells

Differential radioactive quantification of protein abundance ratios between benign and malignant prostate tissues: Cancer association of annexin A3

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