Biosynthesis of Alkaline Phosphatase During Differentiation of the Human Colon Cancer Cell Line Caco-2

Matsumoto, Hisashi; Erickson, Roger H.; Gum, James R.; Yoshioka, Masahiro; Gum, Elizabeth T.; Kim, Young S.

doi:10.1016/0016-5085(90)90334-w

Cited by 155 publications

(113 citation statements)

References 21 publications

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“…These observations and conclusions are consistent with the previous reports (16,(21)(22)(23). The alkaline phosphatase activity, a marker of cell differentiation (24), was clearly increased in the attached cells by the NaBT treatment (Fig. 1B).…”

Section: Apoptosis and Cell Differentiation In Caco-2 Cells Duringsupporting

confidence: 93%

Expression of 15-Lipoxygenase by Human Colorectal Carcinoma Caco-2 Cells during Apoptosis and Cell Differentiation

Kamitani

Geller

Eling

1998

Journal of Biological Chemistry

130

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We studied arachidonic acid metabolism and the expression of cyclooxygenase (Cox) and 15-lipoxygenase (15-LO) in the human colorectal carcinoma cell line, Caco-2, which undergo apoptosis and cell differentiation in the presence of sodium butyrate (NaBT). Caco-2 cells expressed very low levels of Cox-1 but highly expressed Cox-2. NaBT treatment shifted the arachidonic acid metabolites by cell lysates from prostaglandins to 15-hydroxyeicosatetraenoic acid, indicating the presence of a 15-LO. Linoleic acid, an excellent substrate for 15-LO, was metabolized poorly by the Caco-2 cells, but NaBT treatment shifted metabolism to 15-LO metabolite, 13(S)-hydroxyoctadecadienoic acid. Caco-2 cells expressed a 15-LO but only after treatment with NaBT, as determined by Northern blotting. Immunoblotting with anti-human 15-LO antibody detected a 72-kDa band in NaBT-treated Caco-2 cells. Expression of 15-LO mRNA was dependent on the duration of NaBT treatment, with the highest expression observed between 10 and 24 h. Results from expression and metabolism studies with arachidonic and linoleic acid cells indicated Cox-2 was responsible for the lipid metabolism in control cells, whereas 15-LO was the major enzyme responsible after NaBT induction of apoptosis and cell differentiation. The 15-LO in Caco-2 cells was characterized as human reticulocyte 15-LO by reverse transcription-polymerase chain reaction and restriction enzyme analysis. The expression of 15-LO and 15-hydroxyeicosatetraenoic acid or 13(S)-hydroxyoctadecadienoic acid formation correlates with cell differentiation or apoptosis in Caco-2 cells induced by NaBT. The addition of nordihydroguaiaretic acid, a lipoxygenase inhibitor, significantly increased NaBT-induced apoptosis, whereas the addition of indomethacin did not alter NaBT-induced apoptosis in the Caco-2 cells. However, indomethacin treatment decreased the expression of Cox-2 in NaBT-treated cells and significantly increased the expression of 15-LO during NaBT treatment. These studies suggest a role for 15-LO, in addition to Cox-2, in modulating NaBT-induced apoptosis and cell differentiation in human colorectal carcinoma cells.The first genetic alteration in the multistep process that leads to the development of colon carcinogenesis seems to be the loss of APC gene function. Investigations of APC function and its mutations have provided important clues in understanding colon cancer (1). One of the important changes that results from the loss of the APC gene function is the overexpression of prostaglandin H synthase-2 (cyclooxygenase-2, Cox-2), 1 an inducible enzyme that converts arachidonic acid to prostaglandins. The link between APC, Cox-2, and polyp formation was firmly established in studies using APC (Apc ⌬716 ) and Cox-2 knockout mice (2). Mice carrying the APC mutation overexpress Cox-2 and develop intestinal polyps. When bred to mice with the disrupted Cox-2 gene, the offspring, homozygously deficient for Cox-2, had significantly less polyps than the mice with wild-type Cox-2. These results confirmed C...

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Section: Apoptosis and Cell Differentiation In Caco-2 Cells Duringsupporting

confidence: 93%

Expression of 15-Lipoxygenase by Human Colorectal Carcinoma Caco-2 Cells during Apoptosis and Cell Differentiation

Kamitani

Geller

Eling

1998

Journal of Biological Chemistry

130

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“…The development of a brush border was consistent with increased differentiation, known to occur when epithelial cells are grown on permeable filter supports [25,26]. We therefore measured the activities of alkaline phosphatase and sucrase- isomaltase, two enzymes used previously as markers of cell differentiation in this cell type [27]. The increase in specific activity of both enzymes relative to age of cell culture was greatest when parent cells were grown on filters.…”

Section: Figure 1 Sensitivity Of Parent Caco-2 Cells and Caco-2-rca Rmentioning

confidence: 84%

Cell polarization is required for ricin sensitivity in a Caco-2 cell line selected for ricin resistance

Jackman

Ellis

Gray

et al. 1999

Biochemical Journal

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It has been proposed that killing of mammalian cells by ricin requires efficient endocytic delivery to the trans-Golgi network (TGN) prior to retrograde transport to the endoplasmic reticulum and entry to the cytosol. In polarized epithelial cells, an efficient membrane-traffic pathway to the TGN is present from the basolateral but not the apical plasma-membrane domain. Thus one can hypothesize that a ricin-resistant phenotype might be demonstrated by polarized cells that fail to differentiate and thus fail to develop an efficient membrane-traffic pathway from the basolateral plasma membrane to the TGN. We have isolated and studied a ricin-resistant Caco-2 cell clone (Caco-2-RCAr clone 2) which, when grown on plastic, was deficient in differentiation, measured by the development of polarized-cell-surface marker enzymes. The deficiency in differentiation was partially reversed, and ricin sensitivity was restored, when the cells were grown on filter supports. Our data provide the first evidence of a ricin-resistant cell line where resistance is due to the lack of development of polarized cell surfaces. The observed ricin resistance is consistent with the requirement that ricin is delivered to the TGN before its A chain enters the cytosol to mediate cell killing.

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“…Type I collagen was also shown to inhibit expression of the enterocytic marker alkaline phosphatase (Matsumoto et al, 1990) in Caco-2, Colony 29 and HRA-19 cells using an enzymatic method for alkaline phosphatase assay ( Figure 2B). Inhibition of enterocytic differentiation markers was confirmed by measuring intestinal alkaline phosphatase protein expression using a cellbased ELISA.…”

Section: Type I Collagen Inhibits Lineage Differentiation In Human Comentioning

confidence: 99%

Type I collagen inhibits differentiation and promotes a stem cell-like phenotype in human colorectal carcinoma cells

Kirkland

2009

Br J Cancer

126

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BACKGROUND: Human colorectal cancer is caused by mutations and is thought to be maintained by a population of cancer stem cells. Further phenotypic changes occurring at the invasive edge suggest that colon cancer cells are also regulated by their microenvironment. Type I collagen, a promoter of the malignant phenotype in pancreatic carcinoma cells, is highly expressed at the invasive front of human colorectal cancer. METHODS: This study investigates the role of type I collagen in specifying the colorectal cancer cell phenotype. The effect of type I collagen on morphology, localisation of cell -cell adhesion proteins, differentiation and stem cell-like characteristics was examined in a panel of human colorectal carcinoma cell lines. RESULTS: Human colorectal carcinoma cells grown on type I collagen in serum-free medium show an epithelial -mesenchymal-like transition (EMT-like), assuming a more flattened less cohesive morphology. Type I collagen downregulates E-cadherin and b-catenin at cell -cell junctions. Furthermore, type I collagen inhibits differentiation, increases clonogenicity and promotes expression of stem cell markers CD133 and Bmi1. Type I collagen effects were partially abrogated by a function-blocking antibody to a2 integrin. CONCLUSION: Together, these results indicate that type I collagen promotes expression of a stem cell-like phenotype in human colorectal cancer cells likely through a2b1 integrin.

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Biosynthesis of Alkaline Phosphatase During Differentiation of the Human Colon Cancer Cell Line Caco-2

Cited by 155 publications

References 21 publications

Expression of 15-Lipoxygenase by Human Colorectal Carcinoma Caco-2 Cells during Apoptosis and Cell Differentiation

Expression of 15-Lipoxygenase by Human Colorectal Carcinoma Caco-2 Cells during Apoptosis and Cell Differentiation

Cell polarization is required for ricin sensitivity in a Caco-2 cell line selected for ricin resistance

Type I collagen inhibits differentiation and promotes a stem cell-like phenotype in human colorectal carcinoma cells

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