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Biomedical subjects

L H Augenlicht

Publications and source records attributed to L H Augenlicht.

At least 19 recordsLinked to original sources
PubMed1992

Modulation of gene expression as a biomarker in colon.

Computer-driven scanning and image processing methodology has demonstrated that genetic inheritance of risk for colorectal cancer in familial polyposis (FAP) and hereditary non-polyposis colorectal cancer (HNPCC) families is associated with highly pleiotropic effects on patterns of gene expression in the flat colonic mucosa. The mitochondrial (mt) gene encoding subunit 3 of cytochrome oxidase (COXIII) is one of a panel of cloned sequences which characterize genetic risk. Expression of COXIII decreased in progression of, and risk for, colonic tumors in vivo. Further, metabolizable, unbranched, short-chain fatty acids (SCFAs) elevated expression of mtCOXIII, as well as mtCOXI, in HT29 cells and also elevated mtCOX enzymatic activity. However, expression of nuclear encoded COX subunits were unaffected. These changes may be related to documented alterations in mitochondria structure and function in transformed colonic epithelial cells. SCFAs produced when colonic microflora causes fermentation of fiber are the principle energy source for normal colonic epithelial cells; SCFAs also induce a more differentiated phenotype both in vitro and in vivo. Therefore, a mechanistic link may exist between molecular events in inherited risk and a dietary factor (fiber) which may modulate such risk. In a preliminary intervention trial in collaboration with M. Lipkin, high risk HNPCC patients received daily supplements of 1500 mg CaCO3 per day, which may be protective for development of colorectal tumors. Elevations in COXIII expression were seen in 7 of 12 patients within the first 7 months, followed by complex changes in expression of this sequence.

Adenomatous Polyposis Coli↗
PubMed1991

Effects of fatty acids on expression of genes encoding subunits of cytochrome c oxidase and cytochrome c oxidase activity in HT29 human colonic adenocarcinoma cells.

As the primary and preferred energy source of normal colonic epithelial cells, fatty acids may play a unique role in the differentiation and physiology of these cells. We have shown that expression levels of COXIII, a mitochondrial gene encoding one of the 13 subunits of cytochrome c oxidase, are abnormally low in colon tumors and colonic tissue at genetic risk for developing tumors but increase following in vitro treatment of HT29 human colonic adenocarcinoma cells with the fatty acid butyrate. The present studies investigate the specificity of fatty acids in effecting cytochrome c oxidase subunit expression and enzymatic activity in HT29 cells. The data demonstrate that, depending upon their chain length, metabolizable unbranched fatty acids increase expression of two subunits encoded by mitochondrial genes (I and III) and enhance cytochrome c oxidase activity. However, none of the fatty acids had an effect on expression of two subunits encoded by nuclear genes (IV and Va). These findings suggest that the low levels of COXIII expression exhibited in colonic tumors may represent a limiting factor in the assembly of functional cytochrome c oxidase and contribute to the depressed enzyme activity reported in these tumors. By elevating expression of subunits I and III and enzymatic activity, fatty acids may enhance the potential for cellular respiration. The more differentiated phenotype which is reported in colorectal carcinoma cell lines treated with fatty acids in vitro may be, therefore, associated with correction of metabolic abnormalities in transformed cells.

Adenocarcinoma↗
PubMed1991

Patterns of gene expression that characterize the colonic mucosa in patients at genetic risk for colonic cancer.

We have used a computer-driven scanning and image-processing system to identify a panel of 30 cDNA clones whose pattern of expression in individual biopsy specimens distinguishes the flat, normal-appearing colonic mucosa of patients in two genetic groups at high risk for development of colorectal cancer from that of normal colonic mucosa in low-risk individuals. The two high-risk groups, familial adenomatous polyposis and hereditary nonpolyposis colon cancer, are indistinguishable based on the pattern of expression of the 30 selected clones. This suggests that the extensive pleiotropic effects of the inherited loci, which may play an important role in the mechanism of increased risk and early onset of the disease, are similar in these populations.

Adenomatous Polyposis Coli↗
PubMed1991

Aggressive subtypes of human colorectal tumors frequently exhibit amplification of the c-myc gene.

Expression of the c-myc gene is often elevated in human colorectal tumors, but reported amplification of the locus is rare. Here we demonstrate that modest amplification of c-myc is frequently found in aggressive subtypes of colorectal cancer. Careful quantitation of c-myc copy number has shown amplification in 53.8% (7/13) mucinous tumors, 42.3% (3/7) poorly differentiated tumors and a single poorly differentiated APUD tumor. This contrasts with amplification in 6.9% (2/29) moderately to well differentiated tumors, a value which is in agreement with that in previously published reports. Such changes in gene copy number may represent an important aspect of the genomic alterations which accumulate during the development of colorectal tumors.

Blotting, Southern↗
PubMed1990

Expression of mitochondrial cytochrome c oxidase in human colonic cell differentiation, transformation, and risk for colonic cancer.

In a panel of eight cloned complementary DNA sequences whose level of expression characterize colon cells as transformed in vivo and in vitro, one which may also serve as a marker of risk in familial polyposis and familial colon cancer flat mucosa has been identified as mitochondrial cytochrome c oxidase subunit 3. Mean level of expression of cytochrome c oxidase subunit 3 decreases progressively in colon adenomas and carcinomas relative to normal mucosa in vivo, and returns to higher levels present in biopsies of normal mucosa when the HT29 human colonic adenocarcinoma cell line is induced to differentiate with sodium butyrate. Quantitation of cytochrome c oxidase subunit 3 DNA by dot blots indicated that these changes in expression were not associated with alterations in the number of mitochondrial genomes.

Adenomatous Polyposis Coli↗
PubMed1990

Changes in the number of mitochondrial genomes during human development.

Using a cDNA probe for the mitochondrially encoded third subunit of cytochrome c oxidase (COIII) we found a progressive increase in the number of mitochondrial DNA molecules in specific human tissues during normal fetal development. The data indicate that the tissue, rather than the final number of mitochondrial genomes, apparently plays a dominant role in determining the gestational stage at which the adult complement of this DNA is established.

Blotting, Southern↗
PubMed1990

Absence of detectable deletions in the mitochondrial genome of human colon tumors.

Changes in structure, function, and gene expression of mitochondria have been demonstrated in colorectal tumors (Wong, J.R.; Chen, L.B. Adv. Cell Biol. 2:263-290; 1988). In this report, mitochondrial DNA from 30 primary human colorectal tumors was compared to that of adjacent histologically normal colonic mucosa and human placenta. No evidence for substantial deletions in the major or minor population of mitochondrial DNA was detected, despite the fact that there are extensive deletions throughout the nuclear genome of such tumors. Hence, alterations in mitochondria in colorectal tumors must be dependent upon other mechanisms.

Adenocarcinoma, Mucinous↗
PubMed1979

Protected nucleotide sequences in nuclear ribonucleoprotein.

The rapidly labeled nuclear ribonucleic acid in human carcinoma cells which is protected by protein from digestion by staphylococcal nuclease (EC 3.1.4.7) has been investigated. A simple and discrete sequence specificity was not found, but the protected RNA fragments are rich in G + C and were shown by fingerprinting to comprise a nonrandom subset of all heterogeneous nuclear ribonucleic acid (hnRNA) sequences enriched in the sequences AGC, GGC, AGGC, and GAGC. There was no detectable enrichment for dougble-stranded RNA in the protected fraction. These data provide the first evidence that the association of any protein with hnRNA is nonrandom with respect to nucleotide sequence.

Base Sequence↗
PubMed1977

Serum stimulation of human fibroblasts: effect on non-histones of nuclear ribonucleoprotein and chromatin.

In quiescent human fibroblasts stimulated to proliferate by fresh medium plus 15% serum, no changes were seen in the incorporation of 3H tryptophan into the protein of nuclear ribonucleoprotein during the first three hours following re-feeding. This was in contrast to non-histone chromosomal proteins where the incorporation increased by 90% within ten minutes. The density of the formaldehyde fixed nuclear ribonucleoprotein in CsCl was 1.43-1.44 g/ml and this also did not change following stimulation. The electrophoretic profile of the proteins of nuclear ribonucleoprotein on SDS gels exhibited a predominant band corresponding to a molecular weight of 44,000 closely trailed by a band at 47,000 and other bands at higher molecular weight. This pattern was not altered by serum stimulation and the same was true for the more complex electrophoretic profile of the chromatin proteins. Following a 10-minute pulse of 3H-tryptophan at ten minutes after stimulation, there was a selective increase in the labeling of non-histone chromosomal protein of molecular weight 59,000; no change was seen in the labeling of any protein of nuclear ribonucleoprotein.

Blood↗
PubMed1976

Digestion of RNA of chromatin and nuclear ribonucleoprotein by staphylococcal nuclease.

Following a 1-h [3H]uridine pulse of cells of a human colon carcinoma line, 15% of the radioactivity in heterogeneous nuclear RNA associated with both chromatin and nuclear ribonucleoprotein was not digested to acid soluble fragments during a 2-h incubation with staphylococcal nuclease (EC 3.1.4.7). These [3H]uridine-labeled oligonucleotides were approximately 26 nucleotides in length. An RNA containing structure which sedimented no faster than 2 S could be isolated from the digests. Major and mino peptide species, of molecular weights 40 000 and 66 000, respectively, were associated with this structure isolated from either chromatin or nuclear ribonucleoprotein. The results demonstrate that some protein of nuclear ribonucleoprotein is complexed with the transcript while it is still associated with chromatin.

Cell Line↗
PubMed1976

Appearance of rapidly labeled, high molecular weight RNA in nuclear ribonucleoprotein. Release from chromatin and association with protein.

Chromatin and nuclear ribonucleoprotein (nRNP) have been prepared from a human carcinoma cell line. Following a 1-hour (3H)uridine pulse, 60 to 70% of the nuclear radioactivity, after removal of nucleoli, was found in the chromatin, the balance in nRNP. This was true whether the chromatin and nRNP were separated by velocity centrifugation or by isopycnic centrifugation on Metrizamide gradients. Radioactivity in chromatin and nRNP was found in high molecular weight RNA, with mean sedimentation coefficients of 20 S and 15 S, respectively, as determined on sodium dodecyl sulfate-sucrose gradients. Experiments on the kinetics of appearance of radioactivity in the RNA of the two fractions suggest that some of the chromatin-associated RNA is precursor to nRNP-RNA. The proteins of nRNP are complex as revealed by sodium dodecyl sulfate gel electrophoresis. The contamination by chromatin protein was estimated to be 5%. Experiments involving short pulses of (3H)tryptophan, and pulse-chase, suggested that the rapidly turning over proteins of nRNP were not complexed with RNA while still associated with chromatin. However, it was also shown that the radioactivity in nRNP following short pulses of (3H)tryptophan did not correspond to the major bands seen on stained sodium dodecyl sulfate gels. It is therefore concluded that the protein of nRNP consists of two classes: species present in large amounts, possibly common to all RNA in nRNP, which are relatively stable and may be complexed to RNA still associated with chromatin; and a large number of rapidly turning over species, each present in small amounts and associated with nRNP only after its release from chromatin.

Cell Line↗
PubMed1975

Chromosomal proteins of rat brain: increased synthesis and affinity for DNA following a pulse of the carcinogen ethyinitrosourea in vivo.

A single pulse of ethlnitrosourea (EtNU), administered to 10-day-old BD IX-rats, specifically results in a high incidence of neuroectodermal tumors in the central and peripheral nervous system. At five days after an EtNU-pulse, analyses of protein-DNA interactions were performed using chromatin dissociation and re-association experiments, following incorporation of radioactive leucine into brain chromosomal proteins (CP) during short-term suspension culture. In comparison with 15-day-old control animals, the brain cells of EtNU-treated rats exhibited (i) an increased rate of CP synthesis, and (ii) an increased affinity of the newly-synthesized CP for brain DNA of both control and EtNU-treated animals.

Animals↗