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R M Niles

Publications and source records attributed to R M Niles.

At least 37 records · Page 2Linked to original sources
PubMed1994

Interactions between retinoic acid and protein kinase C in induction of melanoma differentiation.

Retinoic acid treatment of B16 mouse melanoma cells induces a differentiated phenotype. This is accompanied by a decrease in monolayer growth rate, loss of the ability to form colonies in soft agarose, increased production of melanin and other melanocyte-specific markers. In addition, retinoic acid treatment of these cells decreases their tumorigenicity when injected subcutaneously into mice. Our laboratory has found that an early biochemical change after the addition of retinoic acid is a large increase in PKC. PKC is an enzyme whose activity is activated by diacylglycerol and calcium and has been shown to be an important mediator of substances that stimulate growth or differentiation. Since PKC is a multi-gene family, it was important for us to determine which isotype(s) was expressed in B16 cells and which type was induced by retinoic acid. We found that only PKC-alpha is expressed in these cells, and this is the form that is induced by retinoic acid. The retinoic acid-induced increased in PKC-alpha is found at both the RNA and protein level. The mechanism of induction is not yet clear since there is only a small increase in the transcription rate and no change in the stability of the mRNA for PKC-alpha in treated cells. In addition, the induction of PKC by retinoic acid can be blocked by inhibitors of protein synthesis, suggesting that the induction requires the synthesis of new protein(s). In order to determine the role of increased PKC-alpha in the retinoic acid-induced differentiation, we transfected full-length PKC-alpha cDNA in mammalian expression vectors into B16 cells. Two clones that stably overexpressed PKC-alpha to different levels were isolated. The phenotype of these clones resembled WT cells treated with retinoic acid, i.e. they had longer doubling times, decreased ability to form colonies in soft agar, increased melanin production, and decreased tumorigenicity in mice. Recent data suggest a role for the RAR-beta in mediating the effect of retinoic acid on PKC induction. B16 cells express a very low amount of RAR-beta mRNA. The level is increased drastically by retinoic acid treatment without any requirement for protein synthesis. When B16 cells were transfected with and overexpressed RAR-beta, they also expressed more PKC-alpha mRNA and protein, and the induction of PKC by retinoic acid was not blocked by protein synthesis inhibitors. In summary, these finding suggest a key role for PKC-alpha in the pathway by which retinoic acid induces B16 mouse melanoma differentiation.

Animals↗
PubMed1994

Expression of TGF-beta during in vitro differentiation of hamster tracheal epithelial cells.

The control of growth and differentiation of tracheal epithelial cells is poorly understood. Retinoic acid seems to be essential for the growth and secretory cell differentiation of hamster tracheal epithelial (HTE) cells in culture. In this study, we tested the hypothesis that one way by which retinoic acid (RA) stimulates growth is by decreasing transforming growth factor beta (TGF beta) expression or activity or both. HTE cells were very sensitive to TGF beta-induced growth inhibition. TGF beta 1 was more potent than TGF beta 2 with 50% inhibition of growth achieved at a concentration less than 0.1 ng/ml. A single TGF beta 1 transcript of 2.4 kb was expressed in HTE cells, and the amount increased by fourfold as cell proliferation decreased and differentiation increased. No TGF beta 2 mRNA could be detected in proliferating undifferentiated HTE cells, but two distinct mRNAs (5.1 and 3.5 kb) were observed to be induced in a transient fashion in RA-treated cells which correlated with the onset of differentiation. The amount of biologically active TGF beta in conditioned media from HTE cells at different stages of growth and differentiation in primary culture was determined by the mink lung epithelial cell growth inhibition assay and the use of neutralizing antibodies. These assays indicated a large increase in the total amount of TGF beta at the time the cells slowed their growth and started to differentiate. The activity was due primarily to TGF beta 1. Interestingly, cells treated with RA had a major component of "preactivated" (non-latent) TGF beta 1 compared to control cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Characterization of human retinoic acid receptor alpha 1 expressed in recombinant baculovirus-infected Sf9 insect cells.

Full-length human retinoic acid receptor alpha 1 (hRAR alpha 1) was expressed in Sf9 insect cells using the baculovirus expression vector system (BEVS). Western blot analysis using a specific anti-RAR peptide antiserum detected two major protein bands with apparent mol wts of approximately 54 and approximately 51 kDa in extracts from insect cells infected with recombinant hRAR alpha 1 Autographica californica (AcNPV) baculovirus. Analysis of recombinant extracts from Sf9 cells labeled in vivo with [32P]orthophosphate suggested that the recombinant protein was phosphorylated. A component in the recombinant nuclear extracts specifically bound [3H]all-trans-retinoic acid (RA) and sedimented in sucrose density gradient centrifugation as a single, symmetric peak with a sedimentation coefficient of approximately 3.6S, corresponding to a protein of approx 50 kDa. Scatchard analyses determined that [3H]RA was bound in recombinant extracts by a single class of binding sites with an apparent dissociation constant of approximately 0.3 nM and nuclear and cytoplasmic extracts contained approximately 1200 and approximately 200 pmoles, respectively, of unoccupied receptor per mg protein. In competitive ligand binding assays, relative binding affinities of 9-cis- and 13-cis-RA for hRAR alpha 1 in nuclear extracts were about threefold and sixfold lower than all-trans-RA, whereas all-trans-retinol, -retinaldehyde, and -retinyl acetate demonstrated relatively weak binding. In gel mobility shift assays, the electrophoretic migration of a [32P]-labeled oligonucleotide containing the retinoic acid response element of the RAR beta gene was retarded in the presence of recombinant nuclear and cytoplasmic extracts. The apparent complex formation between recombinant hRAR alpha 1 and beta RARE was greatly enhanced by the addition of nuclear extract from wild-type AcNPV-infected Sf9 cells, possibly because of heterodimer formation between recombinant hRAR alpha 1 and a metazoan RXR homolog. Thus, recombinant hRAR alpha 1 expressed at high levels in Sf9 insect cells exhibited biochemical properties of the native protein, including nuclear translocation, specific high affinity ligand and RARE binding, and possible heterodimer formation.

Amino Acid Sequence↗
PubMed1993

Quantitative ultrastructural analysis of the relationship between cell growth, shape change, and mucosecretory differentiation in cultured hamster tracheal epithelial cells exposed to retinoic acid.

Hamster tracheal epithelial cells in extended (32 degrees C) primary culture with and without supplemental retinoic acid (RA) were studied during the proliferative (5 days) and differentiation phases (11 days) by correlative transmission electron microscopy (EM) and light microscopic (LM) autoradiography to quantify the relationship between cell proliferation, shape change, and mucin granule expression. In retinyl acetate-containing control medium, cell numerical density was higher and [3H]thymidine labeling index (LI) lower at day 11 compared with day 5. The addition of 10(-7) M RA to the medium caused an increase in cell numerical density at both times. LI was increased by RA at 5 days and decreased at 11 days. Measurements of cell shape in ultrathin sections adjacent to LM autoradiographs made in the vertical plane demonstrated an RA-induced change from flat to cuboidal at 5 days and a more columnar phenotype at 11 days. Cells containing mucin granules were of two main types based on their ultrastructure. One type, seen at 5 and 11 days, contained diminutive mucin granules and had an LI of 50% at 11 days. Its LI and frequency (26%) were unaltered by RA. The other type, less frequent (15%) and present only at 11 days, was more columnar and contained mucous granules similar to those found in vivo. RA doubled the frequency of this cell type but did not affect its LI (11%). Cells of this type with more than five mucin granules in EM profile did not incorporate thymidine. The data indicate that RA accelerates and enhances cell shape change toward a more cuboidal phenotype.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Increased expression of protein kinase C alpha plays a key role in retinoic acid-induced melanoma differentiation.

Differentiation of B16 mouse melanoma cells induced by retinoic acid (RA) is preceded by a large increase in protein kinase C alpha (PKC alpha) mRNA and protein. To determine the role of PKC alpha in the differentiation program, we stably transfected B16-F1 cells with a plasmid containing the full length PKC alpha cDNA driven by an SV40 promoter. Two out of thirty-two colonies screened were determined to overexpress PKC by 2-4-fold according to Western blot analysis and PKC enzyme activity. When compared to control cells (wild-type cells and cells transfected only with the neomycin resistance gene), PKC alpha overexpressing clones displayed longer doubling times, diminished anchorage-independent growth, and increased melanin production. RA treatment of control cells mimicked these phenotypic characteristics. When injected subcutaneously into syngeneic mice, PKC alpha overexpressing clones produced smaller tumors and had longer latencies than control cells. These findings, combined with the fact that phorbol esters down-regulate PKC and antagonize RA action suggest that PKC alpha plays a key role in the RA-induced melanoma differentiation.

Animals↗
PubMed1992

Regulation of protein kinase C gene expression by retinoic acid in B16 mouse melanoma cells.

We have previously shown that the retinoic acid (RA)-induced growth arrest and differentiation of B16 mouse melanoma cells is accompanied by a large increase in the amount and activity of protein kinase C (PKC). Since PKC is a multigene family, we investigated which isoforms were expressed in control and RA-treated B16 melanoma cells, and characterized the manner by which RA regulates PKC gene expression. We found that RA treatment of B16 cells resulted in an increase in PKC alpha mRNA beginning at 4-8 h and reached a maximum of 10- to 12-fold over control levels by 48 h. There was also a small amount of PKC gamma mRNA, present only in 48-h RA-treated cells, but no PKC beta mRNA was detected. The effect of RA on PKC alpha mRNA induction was not direct since the induction was abolished when cycloheximide was included in the incubation medium. Nuclear run-on experiments showed that the RA-induced increase in PKC alpha steady-state mRNA was not entirely due to an increase in transcriptional activity, as the increase in PKC alpha transcription was only 2- to 3-fold over control, which is not enough to account for the 10- to 15-fold increase in steady state levels. There was also no change in PKC alpha mRNA stability in RA-treated B16 cells compared to untreated cells. The 10.9-kb PKC alpha message in both control and RA-treated cells was less stable than the 3.8-kb PKC alpha message. Therefore, we propose that the major level of control of PKC alpha mRNA levels by RA is post-transcriptional, either RNA processing or transport out of the nucleus.

Animals↗
PubMed1991

B16 mouse melanoma cells selected for resistance to cyclic AMP-mediated growth inhibition are cross-resistant to retinoic acid-induced growth inhibition.

B16 mouse melanoma cells are grown inhibited by cyclic AMP or by retinoic acid (RA). However, the combination of these two agents results in less growth inhibition than either agent alone. In order to investigate this interaction, cells were selected for resistance to 8-bromo-cyclic AMP-induced growth inhibition. Two clones (3 and 7) which demonstrated significant resistance were isolated. When these two clones were treated with retinoic acid (RA) it was observed that they also exhibited different degrees of resistance to this growth inhibitor. This cross-resistance did not appear to be due to a lack of uptake or retention of the respective inhibitors, since the mutants took up and retained more 3H-cAMP and 3H-RA than wild type cells, suggesting that the dual resistance was not due to an amplification of P-glycoprotein. The mutation confering cAMP-resistance did not appear to involve cyclic AMP-dependent protein kinase, since both catalytic activity and the amount of cAMP protein binding was similar in wild type and mutants. Thus, the mutation must be beyond the interaction of cAMP with cAMP-dependent protein kinase. We have previously reported that RA induces protein kinase C in B16 melanoma cells (Niles and Loewy: Cancer Res. 49:4483-4487, 1989). Therefore, we measured the ability of RA to induce protein kinase C in the cyclic AMP-resistant mutants. We found an inverse correlation between RA-induced protein kinase C activity and growth inhibition in these mutants. The data reported here suggest that cyclic AMP regulates some step in the RA signal transduction pathway.

Animals↗
PubMed1991

Effects of growth medium and cyclic AMP analogues on the cAMP-induced differentiation of F9 teratocarcinoma cells.

F9 teratocarcinoma cells differentiate into parietal endodermlike cells when treated with retinoic acid (RA) and cyclic AMP (cAMP). We have previously found that F9 cells can be induced to differentiate by treatment with cAMP in the absence of RA. In the course of determining why other investigators had failed to observe cAMP-induced differentiation, we found that the growth medium played an important role in determining the response of F9 cells to differentiating agents. When F9 cells were grown in minimal essential medium (MEM) and treated with either 8-bromo-cAMP (8BrcA) + 1-methyl, 3-isobutylxanthine (MIX), or dibutyryl cAMP (DBcA) + theophylline (T), they differentiated to parietal endodermlike cells without any requirement for exogenous RA. However, when F9 cells were grown in Dulbecco's modified Eagle's medium (DME), DBcA/T failed to induce differentiation alone and required exogenous RA to induce formation of parietal endoderm-like cells. 8BrcA/MIX alone was still able to induce some differentiation, although the extent was not as great as those cells grown in MEM. These results could not be explained by the different growth-promoting properties of the two culture media because there was no difference in the doubling time of F9 cells grown in either medium. Likewise, RA and cAMP both inhibited growth to the same extent in either medium. Inasmuch as almost all published reports on F9 cell differentiation have used DME as a growth medium, and RA with or without DBcA/T as the differentiating agents, these studies would not have had the appropriate conditions to detect the cAMP-induced differentiation of F9 cells.

1-Methyl-3-isobutylxanthine↗
PubMed1990

Cyclic adenosine monophosphate-mediated induction of F9 teratocarcinoma differentiation in the absence of retinoic acid.

F9 teratocarcinoma stem cells differentiate into parietal endoderm-like cells when given retinoic acid (RA) and dibutyryl cyclic adenosine monophosphate (DB-cAMP). It is generally accepted that the stem cells are resistant to the action of cAMP alone and need to be primed by RA in order to respond to cAMP. In this report, we demonstrate that F9 stem cells differentiate into parietal endoderm-like cells in the absence of exogenous RA when treated with cholera toxin and 1-methyl,3-isobutyl xanthine (CT/MIX) or 8-bromo-cAMP/MIX (8B2-cAMP/MIX). Cells treated with CT/MIX or 8B2-cAMP/MIX were morphologically similar to parietal endoderm-like cells, produced high amounts of plasminogen activator, and synthesized both type IV collagen and laminin mRNA. Conversely, markers made in abundance by stem cells such as stage-specific embryonic antigen (SSEA-1) and an mRNA species of 6.8 kb (pST6-135) were markedly reduced in CT/MIX-treated cells. To prove that cAMP alone could induce differentiation Lipidex-1000, a hydrophobic gel, was used to remove 80-90% of the endogenous serum retinoids. F9 cells grown in this retinoid-depleted serum and treated with 8B2-cAMP/MIX differentiated to parietal endoderm-like cells as shown by both dramatic changes in morphology and induction of type IV collagen mRNA. Our results indicate that the differentiation of F9 to parietal endoderm-like cells can be induced by increased intracellular cAMP and is not strictly dependent on the addition of RA.

1-Methyl-3-isobutylxanthine↗
PubMed1990

The effect of retinoic acid on growth and proto-oncogene expression in hamster tracheal epithelial cells.

Retinoic acid (RA) has been shown to be required for the maintenance of epithelial differentiation. Vitamin A deficiency in hamsters induces the tracheal epithelial cells to undergo squamous metaplasia. Reversing the vitamin deficiency restores the tracheal epithelial cells to their normal morphology and function. Using a hamster tracheal epithelial (HTE) cell culture system which undergoes differentiation to predominantly secretory cells in vitro, we found that RA can convert flat, squamous-like cells to compact, cuboidal-like cells, and that it stimulated cell proliferation. The mitogenic response to RA was maximal at 10(-7) M and required at least 48 h of treatment to observe the effect. RNA levels of growth-related genes during the growth and differentiation phases of primary HTE cultures were examined by Northern analysis. RA maintained a high level of c-myc RNA expression in preconfluent cultures, whereas untreated cells had low amounts of c-myc RNA. Expression of RNA for the replication-dependent histone 3.2 followed a similar pattern, i.e., its level was high in retinoid-treated versus control preconfluent cultures. In confluent (fully differentiated) HTE cell cultures, both retinoid-treated and control cells had low RNA levels of c-myc and histone 3.2. c-fos RNA levels were undetectable in either control or treated cells at any stage during primary culture. The RNA level of c-Ha-ras was very low in both control and treated cultures and did not vary with the state of growth or differentiation, except that when RA-treated cultures reached confluence, no c-Ha-ras RNA was detected.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Induction of protein kinase C in mouse melanoma cells by retinoic acid.

Retinoic acid inhibits the proliferation of B16 mouse melanoma cells. It also eliminates the ability of these cells to grow in soft agar. These biological actions of retinoic acid have been shown to be accompanied by an increase in the amount of cyclic AMP-dependent protein kinase and an induction of a new isozyme form (RII beta). In this report we demonstrated that retinoic acid-treated B16 melanoma cells had large increases in protein kinase C activity. This increased enzyme activity was accompanied by increases in both the number of phorbol dibutyrate binding sites and the amount of immunoreactive protein kinase C. Other treatments (melanocyte-stimulating hormone, serum deprivation) which inhibited the growth of these cells did not increase protein kinase C activity. When B16 melanoma cells were treated for a prolonged time (72 h) with phorbol dibutyrate, protein kinase C activity was barely detectable. Under these conditions, melanin production was inhibited and cell growth was accelerated. When retinoic acid was added together with phorbol dibutyrate, it prevented the growth stimulatory effect of the phorbol ester and increased protein kinase C activity. However, the absolute activity of the enzyme was still below that found in control cells and very much lower than in cells treated with retinoic acid alone. Taken together with our previous findings, we propose that the increase in protein kinase C might be part of a differentiation program induced by retinoic acid.

Blotting, Western↗
PubMed1989

Sodium butyrate suppresses the transforming activity of an activated N-ras oncogene in human colon carcinoma cells.

The transforming activity of DNA from a newly established undifferentiated human colon carcinoma cell line (MIP-101) was tested in the NIH-3T3 transfection assay. Southern blot analysis of the transfectant DNA revealed the presence of a human N-ras oncogene. Treatment of MIP-101 cells with the maturational agent sodium butyrate induced a more normal phenotype, including diminished growth rate, elimination of anchorage independent growth, and decreased tumorigenicity (R. Niles, S. Wilhelm, P. Thomas, and N. Zamcheck (1988) J. Cancer Invest. 6, 39). Here we report that there is a significant reduction in the transforming efficiency of the DNA from butyrate-treated MIP-101 cells. A nonspecific reduction in total DNA uptake as an explanation for these findings was eliminated by showing that there was similar uptake and expression of the thymidine kinase gene from the DNA of butyrate-treated and control MIP cells. Butyrate treatment had no detectable effect on the overall structure, methylation, and level of expression of the human N-ras gene from MIP-101 cells. An NIH-3T3 transformant ability after treatment with sodium butyrate. Although butyrate suppressed several transformed properties similar to MIP-101 cells, DNA from control and treated cultures had an identical level of transforming activity. The results suggest that the environment of the MIP cells may contain additional elements not present in the NIH-3T3 transformants which are required to observe the effect of butyrate on reduction of transforming activity.

Butyrates↗
PubMed1989

Human neutrophil elastase causes glycoconjugate release from the epithelial cell surface of hamster trachea in organ culture.

It is known that human neutrophil elastase (HNE) treatment of hamster tracheal explants causes the release of glycoconjugates, most of which appear to have the characteristics of mucus glycoproteins. This study was designed to determine the origin of HNE-induced glycoconjugate release from 1-day-old cultures of adult hamster trachea. After confirming that HNE treatment released glycoconjugates from cultures labeled with tritiated glucosamine, light microscopic autoradiograms and electron micrographs were prepared. Untreated cultures and cultures treated with inactivated HNE served as controls. HNE treatment caused a 40 to 50% decrease in the silver grain count on the external apical surfaces of secretory cells (p less than 0.05) and ciliated cells (p less than 0.01). Silver grain counts in secretory and ciliated cell cytoplasm, submucosa, and nontissue background were not significantly different from controls. The percentage of nongranulated secretory cells and the number of secretory granules in granulated secretory cells were similar in the HNE-treated and untreated controls. There was no evidence of constitutive release of radiolabeled glycoproteins, or of discharge of secretory granules from the secretory cells. We conclude that HNE releases mucins and other glycoconjugates from the external surfaces of both secretory and ciliated cells in tracheal organ culture.

Animals↗
PubMed1988

Membrane differentiation markers of airway epithelial secretory cells.

We describe here a system for culturing epithelial cells isolated from hamster trachea, which results in a highly enriched population of mucus-secreting cells. The culture system has enabled us to study the process of secretory cell differentiation in vitro. We found that epithelial secretory cells, in vivo and after 5 days in vitro, selectively bind the lectin Helix pomatia agglutinin (HPA) to apical and, to a lesser extent, basolateral surfaces as well as to mucin granules and intracellular secretory organelles. SDS-PAGE gels of detergent extracts of secretory cells cultured for 5 days reveal three HPA-binding glycoproteins with MW of 120 KD, 220 KD, and greater than 400 KD. The high-MW glycoprotein appears identical to mucin, since it is found in secretions from intact trachea and in spent media from 5-day cultures. It does not appear in spent media from 3-day cultures when cells contain few mucous granules and secrete little mucin. The 220 KD HPA-binding glycoprotein is also present in 5-day but not in 3-day cultures. In contrast, the 120 KD glycoprotein is present at both times. HPA-gp120 is a hydrophobic integral membrane protein, whereas HPA-gp220 and mucin are hydrophilic and are membrane associated. These studies define three membrane glycoproteins, one of which is specific for the tracheal epithelial secretory cell regardless of its mucous content, whereas the other two glycoproteins correlate with mucin secretion. They also demonstrate that, in the fully differentiated state, mucin is bound in a non-covalent fashion to the apical plasma membrane of the tracheal epithelial secretory cell.

Animals↗
PubMed1988

The effect of sodium butyrate and retinoic acid on growth and CEA production in a series of human colorectal tumor cell lines representing different states of differentiation.

The effect of sodium butyrate and retinoic acid added singly or in combination on substrate-dependent growth, colonization efficiency in soft agar, and carcino-embryonic antigen (CEA) production in three human colorectal carcinoma cell lines differing in their degree of differentiation was studied. All three colon cancer cell lines regardless of their state of differentiation had their growth markedly slowed by sodium butyrate, and to a lesser extent by retinoic acid. When both agents were added together, a small synergistic inhibition of growth was noted in all the cell lines. Butyrate eliminated colony formation in soft agar in all three cell lines, however, retinoic acid only reduced colony formation in the well differentiated cell line DLD-2. Sodium butyrate was able to induce CEA production in the undifferentiated cell (MIP-101) and the moderately differentiated cells (clone D) which were previously negative for this marker. It also enhanced the baseline production of CEA in the well differentiated cells (DLD-2). Retinoic acid did not induce CEA production in clone D or MIP-101 cells, but did enhance the production of CEA in DLD-2 cells. When both retinoic acid and sodium butyrate were added together, CEA production was either additive (DLD-2) or was inhibited (clone D and MIP-101). One explanation of these results is that only well differentiated cells have functional cellular retinoic acid-binding protein (cRABP), and that certain actions of retinoic acid (inhibition of anchorage-dependent growth) are independent of the presence of cRABP.

Butyrates↗
PubMed1987

Human neutrophil elastase releases cell surface mucins from primary cultures of hamster tracheal epithelial cells.

Primary hamster tracheal epithelial cells growing on a collagen gel matrix produce high molecular weight mucins indistinguishable from mucins produced in vivo. Using a modified version of these confluent cultures, we have demonstrated here that (i) release of mucins can be stimulated by human neutrophil elastase (HNE; EC 3.4.21.37); (ii) HNE can degrade mucins, and both mucin release and degradation by HNE require an active catalytic site; and (iii) there are at least two pools of mucins in these cells: one is a rapidly turning-over spontaneously releasable constitutive pool, the other is a slowly turning-over HNE-releasable pool. We provide evidence that the HNE-releasable mucins are membrane bound and associated with the secretory cell apical surface.

Animals↗
PubMed1987

Isolation and characterization of an undifferentiated human colon carcinoma cell line (MIP-101).

An undifferentiated human colon carcinoma cell line was established from tumor tissue obtained from metastasis to the liver of colonic adenocarcinoma in a patient with fulminant Dukes D colorectal carcinoma. Histological analysis of the tumor biopsy from the liver confirmed the hospital pathology report of poorly differentiated colonic adenocarcinoma. Explants of this tumor tissue xenografted into a nude mouse were used to establish an epithelioid-like cell culture line, MIP-101. The cell line formed tumors in nude mice that histologically appeared undifferentiated and did not stain for carcinoembryonic antigen (CEA). No CEA was present either by radioimmunoassay (RIA) of the culture supernatant or by immunoperoxidase staining of the tumors or monolayers. MIP-101 appears to be one of the most undifferentiated human colon carcinoma cells lines available. It should prove useful in the search for markers of undifferentiated colonic cancer and in studies of colonic cancer differentiation.

Adenocarcinoma↗