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J A Moshier

Publications and source records attributed to J A Moshier.

31 records · Page 2Linked to original sources
PubMed1992

Polyamine levels, ornithine decarboxylase (ODC) activity, and ODC-mRNA expression in normal and cancerous human colonocytes.

Ornithine decarboxylase (ODC) and polyamines (putrescine, spermidine, and spermine) are crucial for cell proliferation. Recently, elevated ODC activity and polyamine levels have been suggested as biological markers for human colon cancer. In this study, we measured ODC activity and the levels of polyamines (putrescine, spermidine, spermine, and cadaverine) and acetyl-putrescine in human colonocytes isolated from cancerous areas compared to the adjacent normal colon tissue. In addition, ODC mRNA expression was compared between both groups. We found that colonocytes isolated from cancerous areas had significantly higher mean value of ODC activity, putrescine, spermidine, spermine, and cadaverine levels up to 1480%, 470%, 260%, 380%, and 510% respectively compared to colonocytes isolated from the adjacent normal colonic mucosa. No difference was found in acetyl-putrescine levels between cancerous and normal colonocytes. Steady-state levels of ODC mRNA were slightly elevated in cancerous colonocytes relative to normal colonocytes in two of three paired samples. However, the increase in ODC mRNA levels is not sufficient to account for the increase in ODC activity suggesting that colonocyte ODC activity is regulated post-transcriptionally.

Adult↗
PubMed1992

Detection of ornithine decarboxylase messenger RNA in human hepatocellular carcinoma by in situ hybridization.

Ornithine decarboxylase (ODC) has been shown by biochemical analysis, to be important for cell proliferation and carcinogenesis in a variety of tissues, including the liver. We detected messenger RNA (mRNA) specific for the enzyme ODC in 18 patients with hepatocellular carcinoma by an in situ hybridization technique using a radiolabelled ODC probe on formalin-fixed liver specimens. Adjacent uninvolved liver tissues were used as controls. Among the adjacent uninvolved liver tissues, five showed evidence of cirrhosis. Poorly differentiated hepatocellular carcinoma has significantly higher levels of ODC mRNA than does well-differentiated hepatocellular carcinoma, which in turn has a significantly higher ODC mRNA level than adjacent uninvolved liver tissues; tissues showing evidence of cirrhosis, on the other hand, had a significantly lower ODC mRNA level than adjacent uninvolved liver tissue. This pattern of ODC gene expression in hepatocellular carcinoma is similar to the pattern of expression of other oncogenes in liver tumours. The quantitative detection of ODC mRNA in hepatocellular carcinoma by in situ hybridization may help elucidate the potential role of ODC in hepatocarcinogenesis.

Carcinoma, Hepatocellular↗
PubMed1991

Effect of gastric mucosal injury on ornithine decarboxylase in young and aged rats.

The present investigation examines the changes in ornithine decarboxylase (ODC) activity, level of the enzyme, and the expression of its gene in gastric mucosa of young (4-month) and aged (24-month) Fischer-344 male rats 6 h after intragastric administration of either 2 M NaCl (1 ml/130 g b.w.) or an equivalent volume of water (controls). In addition, electronmicroscopy was performed to evaluate the ultrastructural changes in the gastric mucosa. Although administration of 2 M NaCl virtually eliminated the surface epithelium in both young and aged rats, the extent to injury in older animals extended beyond the surface epithelium. In aged rats, epithelial cells in the deeper parts of the gastric glands demonstrated severe swelling with vacuolization and disintegration of the cell organelles, with dying and dead cells. Basal gastric mucosal ODC activity (data from the controls) in aged rats was found to be 118% (p less than 0.001) above the young animals. This was also associated with similar increases in the concentration of ODC (as determined by Western-blot analysis) and a steady-state rise in ODC mRNA. Intragastric administration of 2 M NaCl (which caused gastric mucosal injury) resulted in a 625% increase in mucosal ODC activity in young rats, but in aged rats it produced a 112% increase when compared with the corresponding controls. In young rats, the increase in gastric mucosal ODC activity after injury was also associated with about a 2-fold rise in the enzyme protein concentration and a 4-fold increase in steady-state ODC mRNA levels. In contrast, gastric mucosal injury in aged rats, which resulted in a 112% increase in ODC activity, produced about a 30% reduction in the concentration of ODC and a 15-20% reduction in steady-state mRNA levels, when compared with the respective controls. The current data demonstrate that aging is associated with decreased responsiveness of gastric mucosal ODC to injury which may in part be responsible for diminished regenerative capacity of the gastric mucosa in aged animals. Furthermore, in aged rats the injury-induced stimulation of mucosal ODC activity is not associated with increased activation of its gene.

Aging↗
PubMed1990

Isolation and expression of a human ornithine decarboxylase gene.

We have isolated a human ornithine decarboxylase (ODC) gene from a leukocyte genomic DNA library in order to examine the mechanisms involved in the regulation of ODC gene expression in normal and neoplastic cell growth. Nucleotide sequence analysis shows that the human ODC gene in clone ODC709-A2 consists of 12 exons which encode a protein identical to that inferred from a human ODC cDNA sequence. The 5' end of the gene was determined by S1 nuclease and primer extension mapping. The high G + C content and small open reading frame found in exon 1 may be pertinent to translation regulation of ODC. Conserved sequences and potential promoter elements including a TATA box, a possible CCAAT element, SP1 and AP-2 transcription factor binding sites, and cAMP response elements were identified in the 5'-flanking region. Transfection of mouse LM (tk-) cells with ODC709-A2 DNA resulted in the production of human ODC mRNA approximately 2.25 kilobases in length. Evidence that the protein synthesized from the human gene is functional is provided by "rescue" transfection of a Chinese hamster ovary mutant cell line, C55.7, which is ODC-deficient. C55.7 cells transfected with ODC709-A2 DNA expressed ODC enzyme activity and proliferated without exogenous putrescine.

Animals↗
PubMed1990

Gastrin and epidermal growth factor induction of ornithine decarboxylase in rat colonic explants.

An organ culture system was utilized to examine the effect of gastrin (G-17-I) and epidermal growth factor (EGF) on colonic mucosal ornithine decarboxylase (ODC) activity, and the expression of the ODC gene. Exposure of colonic mucosal explants to either gastrin or EGF (50-500 ng/ml) for only 4 h resulted in a profound stimulation (150-600%) in ODC activity over the basal level. These increases were essentially abolished by difluoromethylornithine (DFMO; 2 nmol/ml) or CaCl2 (2 umol/ml). Gastrin also activated the ODC gene in the colonic mucosa as evidenced by increased steady-state ODC mRNA levels in the colonic mucosal explants after 4 h exposure to the hormone, when compared with the controls. It is concluded that colonic mucosal ODC is responsive to both gastrin and EGF.

Animals↗
PubMed1989

Biochemical changes in the gastric mucosa after injury in young and aged rats.

Changes in gastric mucosal thymidine kinase (TK) activity (an indicator of proliferative activity) were examined in young (4 month) and aged (24 month) Fischer-344 male rats 6 h after intragastric administration of either 2 M NaCl (1 ml/130 g b.w.) or an equivalent volume of water (control). These changes were related to the expression of c-myc gene, tyrosine kinase (Tyr-K) activity and tyrosine-specific phosphorylation of proteins in the gastric mucosa. Basal gastric mucosal TK activity (data from the controls) in the aged rats was found to be 75% (P less than 0.001) above the young animals. This was accompanied by increased expression of c-myc gene and a 67% (P less than 0.001) enhancement in Tyr-K activity. Intragastric administration of 2 M NaCl resulted in gastric mucosal damage (as evidenced by lesions index) in both age groups. However, in aged rats, the lesions index was found to be about 75% higher than in their younger counterparts. In young rats, mucosal injury resulted in a 95% rise in TK activity, whereas in aged rats it was increased by only 38%, when compared with corresponding controls. This 2-fold rise in TK activity in young rats was also associated with increased expression of the c-myc gene. In young rats, administration of hypertonic saline caused a 90% (P less than 0.001) increment in Tyr-K activity and significantly stimulated tyrosine-specific phosphorylation of five mucosal proteins with an apparent molecular mass of 170, 120, 100, 55 and 43 kDa. On the other hand, administration of hypertonic saline to the aged rats caused only a small 16% (P less than 0.025) increase in Tyr-K activity, and produced no apparent change in either expression of c-myc gene or tyrosine-specific phosphorylation of any of the proteins in the gastric mucosa, when compared with the corresponding controls. We conclude that aging increases the susceptibility of the gastric mucosa to damaging agents and diminishes its regenerative capacity. We also suggest that Tyr-K may play a role in determining these events.

Aging↗
PubMed1988

Ornithine decarboxylase as a marker for colorectal polyps and cancer.

There is as yet no specific chromosomal abnormality or gene marker identified for colorectal polyps and cancer. Thus available markers include only phenotypic markers. Tumor markers that have been studied include tetraploidy and increased colonic mucosal proliferation; and these markers have identified those patients that are at high risk for colon cancer. The current "gold standard" of colorectal cancer markers is the carcinoembryonic antigen (CEA). CEA is best used as a monitor of disease and recurrence, and not as a screening or diagnostic test. Newer carbohydrate markers include CA 19-9, incompatible A and B antigens, and T and Lewis antigens. These markers have not shown increased specificity or sensitivity compared to CEA. An interesting recently described marker is ornithine decarboxylase (ODC), which serves as a simple overall index of colonic mucosal proliferation. Ornithine decarboxylase levels have shown correlation with the progression from normal mucosa to adenoma and carcinoma, especially in hereditary polyposis syndromes. This enzyme may also serve as a potential therapeutic target. Many markers have been found useless in further clinical trials. Ornithine decarboxylase needs to be studied in greater detail to determine its sensitivity and specificity, in patients with hereditary colonic neoplasia and in patients without genetic syndromes.

Biomarkers, Tumor↗
PubMed1988

Biochemical markers in colorectal cancer: diagnostic and therapeutic implications.

Markers that are now in use, including CEA and CA-19-9, are not specific or sensitive enough to detect early colorectal cancer. Newer tumor markers such as polyamines, ornithine decarboxylase, and altered blood group carbohydrate antigens may have a potential as future tumor markers. Additional studies of these markers as well as the development of new biochemical markers are warranted in the future to enhance the sensitivity and specificity of diagnosis of early colorectal cancer and those at risk for developing cancer. Finally, understanding events involved in abnormal cell proliferation (that is, elevated polyamines and ODC in colorectal cancer) may help direct future chemotherapy and possibly chemoprevention in high-risk groups such as adenomatous polyposis coli.

Antibodies, Monoclonal↗
PubMed1987

Structure and in vitro transcription of a mouse B1 cluster containing a unique B1 dimer.

A highly repetitive DNA element located 950 bp upstream from a mouse U2 small nuclear RNA gene has been cloned and characterized. The repetitive element is composed of a simple sequence repeat and a cluster of three B1 sequences. Two of these B1 elements are arranged head-to-tail and are joined by an oligo(dA)-rich linker. This unique B1 dimer, comprised of 339 bp, resembles the dimeric structure of primate Alu-family sequences, particularly that of a prototypic human Alu element. The other B1 element within the mouse cluster is a typical monomeric unit. Transcription studies performed in HeLa cell extracts with deletion mutants of the B1 cluster reveal that the single B1 unit is expressed at least 50 times more efficiently than the B1 dimer region. Furthermore, the B1 dimer which contains mutations in the first polymerase III promoter region is not transcribed end-to-end. We conclude that this B1 dimer is unlikely to give rise to a new dimeric retroposon family in the mouse genome.

Animals↗
PubMed1986

mRNA from human colon tumor and mucosa related to the pol gene of an endogenous A-type retrovirus.

The expression of endogenous retroviral genes in mammals may be etiologically related to genetic diseases including cancer. Recently, A-type human endogenous retroviral genomes have been cloned using the reverse transcriptase (pol) genes of rodent intracisternal A-particles (IAP). In this report, RNA from human colon adenocarcinoma and surrounding mucosa was hybridized to mouse IAP pol and gag genes to examine the expression of human endogenous A-type retroviruses. Abundant, heterogeneous size, polyadenylated transcripts homologous to the mouse IAP pol gene were detected in both tissues. Transcripts homologous to the mouse IAP gag region were not found.

Adenocarcinoma↗
PubMedDate not supplied

Expression of protease genes in the gastric mucosa during aging.

The current investigation examines the changes in the expression of pepsinogen C and cathepsin D and E genes in the gastric mucosa during aging and following physiological stimuli of fasting and refeeding. Northern blot analysis of gastric mucosal RNA, isolated from overnight fasted 6-, 12-, and 24-month-old male Fischer 344 rats, revealed that although steady-state mRNA levels of each of these protease remained essentially unchanged between 6 and 12 months of age, in 24-month-old rats the levels were decreased by about 60%, when compared with their younger counterparts. Interestingly, the relative concentration of beta-actin mRNA--but not 18s rRNA--in 12- and 24-month-old rats was also decreased by 23% and 37%, respectively, when compared with 6-month-old animals. In the next set of experiments, groups of young (3 month) and aged (24 month) rats were either fed throughout (controls) or fasted for 48 h and then fed for 6 h and 24 h. Gastric mucosal RNA from each group was assayed for steady-state mRNA levels of pepsinogen C and cathepsin D. Results showed that whereas in young rats fasting decreased pepsinogen C and cathepsin D mRNAs by 80-85%, in aged rats only pepsinogen mRNA was significantly decreased (45%), when compared with the corresponding initial fed controls. In both age groups, refeeding increased pepsinogen C mRNA concentration essentially to the respective initial fed levels. In contrast, cathepsin D mRNA levels in the gastric mucosa of aged rats was affected neither by fasting nor by refeeding. Our current data show that aging not only diminishes the expression of protease genes in the gastric mucosa, but also the expression of one of its structural genes, beta-actin. In addition, responsiveness of these protease genes to the physiological stimuli of fasting and refeeding is also attenuated by aging. We postulate that these age-related changes may in part be due to diminished differentiation of gastric mucosal cells.

Actins↗
PubMedDate not supplied

Androgen regulation of the human ornithine decarboxylase promoter in prostate cancer cells.

We studied the response of the human ornithine decarboxylase (ODC) promoter to androgen in human prostate cancer cell lines. In the well-differentiated, androgen-sensitive human prostate cancer line LNCaP, a genomic ODC promoter fragment that includes putative androgen response elements was suppressed by androgen. In contrast, the androgen-regulated probasin promoter was induced by androgens. The ODC promoter was also induced by cotransfected androgen receptor in the poorly differentiated, androgen-insensitive human prostate cancer cell line PPC-1. We examined the effects of cotransfected mutant androgen receptors containing the LNCaP mutation or DNA-binding mutations. All cotransfected androgen receptors switched the ODC androgen response from suppression to induction in LNCaP cells. Gel-shift and DNA footprint assays demonstrated androgen receptor binding to an ODC sequence that does not contain a consensus androgen response element. Deletion of the sequence abolished androgen suppression of the ODC promoter. We propose a model of pleiotropic gene regulation by androgen that requires a regulatory balance between androgen receptor and a transcription factor binding to the nonconsensus androgen response element.

Androgens↗