Activity and properties of α- l-fucosidase are dependent on the state of enterocytic differentiation of HT-29 colon cancer cells
Previously we have demonstrated an impairment in the activity of α- l-fucosidase in colon tumours. In order to establish an in vitro model to study this enzyme in colon cancer, we have determined the activity and properties of the enzyme during the differentiation of HT-29 colon cancer cells. Cultur...
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| Veröffentlicht in: | The international journal of biochemistry & cell biology 2002-10, Vol.34 (10), p.1291-1303 |
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| container_title | The international journal of biochemistry & cell biology |
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| creator | Merino-Trigo, Ana Rodrı́guez-Berrocal, Francisco Javier de Miguel, Encarnación Páez de la Cadena, Marı́a |
| description | Previously we have demonstrated an impairment in the activity of α-
l-fucosidase in colon tumours. In order to establish an in vitro model to study this enzyme in colon cancer, we have determined the activity and properties of the enzyme during the differentiation of HT-29 colon cancer cells.
Cultures were committed to differentiate into enterocyte-like cells by placing them in a culture medium without glucose for 18–21 days. The state of differentiation was evaluated by assaying the activity of enterocytic marker enzymes, and the acquisition of enterocyte morphology was assessed by electron microscopy. The α-
l-fucosidase activity was determined using a fluorometric method.
Intracellular levels of α-
l-fucosidase activity are lower in non-differentiated cells (3.0±1.01
U/mg) than in differentiated ones (9.2±4.09
U/mg) (
P |
| doi_str_mv | 10.1016/S1357-2725(02)00067-5 |
| format | Article |
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l-fucosidase in colon tumours. In order to establish an in vitro model to study this enzyme in colon cancer, we have determined the activity and properties of the enzyme during the differentiation of HT-29 colon cancer cells.
Cultures were committed to differentiate into enterocyte-like cells by placing them in a culture medium without glucose for 18–21 days. The state of differentiation was evaluated by assaying the activity of enterocytic marker enzymes, and the acquisition of enterocyte morphology was assessed by electron microscopy. The α-
l-fucosidase activity was determined using a fluorometric method.
Intracellular levels of α-
l-fucosidase activity are lower in non-differentiated cells (3.0±1.01
U/mg) than in differentiated ones (9.2±4.09
U/mg) (
P<0.001). This variation is not due to a greater secretion of the enzyme to the culture medium, and properties such as pH optimum or the affinity towards substrate are not dependent on differentiation. The enzyme however, is more stable at acidic pH and at high temperatures, and
V
max is higher in differentiated cells. Moreover, in undifferentiated cells the enzyme is mainly in a monomeric form whereas multimeric forms of the enzyme appear only upon differentiation.
Most of these changes are very similar to those previously observed between normal colon tissue and colon tumours. Thus, we suggest that differentiation of HT-29 colon cancer cells could be used as a model to study the alterations of the enzyme α-
l-fucosidase during the progression of the tumoural process.</description><identifier>ISSN: 1357-2725</identifier><identifier>EISSN: 1878-5875</identifier><identifier>DOI: 10.1016/S1357-2725(02)00067-5</identifier><identifier>PMID: 12127580</identifier><language>eng</language><publisher>Netherlands: Elsevier Ltd</publisher><subject>alpha-L-Fucosidase - metabolism ; Blotting, Western ; Cell Differentiation ; Chromatography, Gel ; Colonic Neoplasms - enzymology ; Colonic Neoplasms - pathology ; Colonic Neoplasms - ultrastructure ; Electrophoresis, Polyacrylamide Gel ; Enzymes ; HT-29 colon cancer cells ; HT29 Cells ; Humans ; Microscopy, Electron ; α- l-Fucosidase</subject><ispartof>The international journal of biochemistry & cell biology, 2002-10, Vol.34 (10), p.1291-1303</ispartof><rights>2002 Elsevier Science Ltd</rights><rights>Copyright 2002 Elsevier Science Ltd.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c361t-e6c70150e7bedd5be0a9463f03cce937c65fc8d73a0f0f3cbe14a52e0a7670653</citedby><cites>FETCH-LOGICAL-c361t-e6c70150e7bedd5be0a9463f03cce937c65fc8d73a0f0f3cbe14a52e0a7670653</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S1357272502000675$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12127580$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Merino-Trigo, Ana</creatorcontrib><creatorcontrib>Rodrı́guez-Berrocal, Francisco Javier</creatorcontrib><creatorcontrib>de Miguel, Encarnación</creatorcontrib><creatorcontrib>Páez de la Cadena, Marı́a</creatorcontrib><title>Activity and properties of α- l-fucosidase are dependent on the state of enterocytic differentiation of HT-29 colon cancer cells</title><title>The international journal of biochemistry & cell biology</title><addtitle>Int J Biochem Cell Biol</addtitle><description>Previously we have demonstrated an impairment in the activity of α-
l-fucosidase in colon tumours. In order to establish an in vitro model to study this enzyme in colon cancer, we have determined the activity and properties of the enzyme during the differentiation of HT-29 colon cancer cells.
Cultures were committed to differentiate into enterocyte-like cells by placing them in a culture medium without glucose for 18–21 days. The state of differentiation was evaluated by assaying the activity of enterocytic marker enzymes, and the acquisition of enterocyte morphology was assessed by electron microscopy. The α-
l-fucosidase activity was determined using a fluorometric method.
Intracellular levels of α-
l-fucosidase activity are lower in non-differentiated cells (3.0±1.01
U/mg) than in differentiated ones (9.2±4.09
U/mg) (
P<0.001). This variation is not due to a greater secretion of the enzyme to the culture medium, and properties such as pH optimum or the affinity towards substrate are not dependent on differentiation. The enzyme however, is more stable at acidic pH and at high temperatures, and
V
max is higher in differentiated cells. Moreover, in undifferentiated cells the enzyme is mainly in a monomeric form whereas multimeric forms of the enzyme appear only upon differentiation.
Most of these changes are very similar to those previously observed between normal colon tissue and colon tumours. Thus, we suggest that differentiation of HT-29 colon cancer cells could be used as a model to study the alterations of the enzyme α-
l-fucosidase during the progression of the tumoural process.</description><subject>alpha-L-Fucosidase - metabolism</subject><subject>Blotting, Western</subject><subject>Cell Differentiation</subject><subject>Chromatography, Gel</subject><subject>Colonic Neoplasms - enzymology</subject><subject>Colonic Neoplasms - pathology</subject><subject>Colonic Neoplasms - ultrastructure</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzymes</subject><subject>HT-29 colon cancer cells</subject><subject>HT29 Cells</subject><subject>Humans</subject><subject>Microscopy, Electron</subject><subject>α- l-Fucosidase</subject><issn>1357-2725</issn><issn>1878-5875</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEtuHCEQhlEUK34kR3DEKooXxDyGpmdlWZYfkSxlYWeNmKJQsHqaCTCWZukj5SI-U2jPWF56BRQf_FUfIceC_xBcdKd3QmnDpJH6O5cnnPPOMP2BHIje9Ez3Rn9s-1dknxyW8tAgoaX6RPaFFNLonh-Qp3Oo8THWDXWjp6ucVphrxEJToM__GB1YWEMq0buC1GWkHlc4ehwrTSOtf5CW6ipOeKthTrCpEaiPIWBulehqbGC7vrlnck4hDe0IbgTMFHAYymeyF9xQ8MtuPSK_ry7vL27Y7a_rnxfntwxUJyrDDkxrn6NZoPd6gdzNZ50KXAHgXBnodIDeG-V44EHBAsXMadkw0xneaXVEvm3_bUP-XWOpdhnL1IEbMa2LNWKu-pmSDdRbEHIqJWOwqxyXLm-s4HZyb1_c20ms5dK-uLdTwNddwHqxRP_2aie7AWdbANuYjxGzLRCxmfAxI1TrU3wn4j-lK5Xb</recordid><startdate>20021001</startdate><enddate>20021001</enddate><creator>Merino-Trigo, Ana</creator><creator>Rodrı́guez-Berrocal, Francisco Javier</creator><creator>de Miguel, Encarnación</creator><creator>Páez de la Cadena, Marı́a</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20021001</creationdate><title>Activity and properties of α- l-fucosidase are dependent on the state of enterocytic differentiation of HT-29 colon cancer cells</title><author>Merino-Trigo, Ana ; Rodrı́guez-Berrocal, Francisco Javier ; de Miguel, Encarnación ; Páez de la Cadena, Marı́a</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c361t-e6c70150e7bedd5be0a9463f03cce937c65fc8d73a0f0f3cbe14a52e0a7670653</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>alpha-L-Fucosidase - metabolism</topic><topic>Blotting, Western</topic><topic>Cell Differentiation</topic><topic>Chromatography, Gel</topic><topic>Colonic Neoplasms - enzymology</topic><topic>Colonic Neoplasms - pathology</topic><topic>Colonic Neoplasms - ultrastructure</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzymes</topic><topic>HT-29 colon cancer cells</topic><topic>HT29 Cells</topic><topic>Humans</topic><topic>Microscopy, Electron</topic><topic>α- l-Fucosidase</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Merino-Trigo, Ana</creatorcontrib><creatorcontrib>Rodrı́guez-Berrocal, Francisco Javier</creatorcontrib><creatorcontrib>de Miguel, Encarnación</creatorcontrib><creatorcontrib>Páez de la Cadena, Marı́a</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The international journal of biochemistry & cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Merino-Trigo, Ana</au><au>Rodrı́guez-Berrocal, Francisco Javier</au><au>de Miguel, Encarnación</au><au>Páez de la Cadena, Marı́a</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Activity and properties of α- l-fucosidase are dependent on the state of enterocytic differentiation of HT-29 colon cancer cells</atitle><jtitle>The international journal of biochemistry & cell biology</jtitle><addtitle>Int J Biochem Cell Biol</addtitle><date>2002-10-01</date><risdate>2002</risdate><volume>34</volume><issue>10</issue><spage>1291</spage><epage>1303</epage><pages>1291-1303</pages><issn>1357-2725</issn><eissn>1878-5875</eissn><abstract>Previously we have demonstrated an impairment in the activity of α-
l-fucosidase in colon tumours. In order to establish an in vitro model to study this enzyme in colon cancer, we have determined the activity and properties of the enzyme during the differentiation of HT-29 colon cancer cells.
Cultures were committed to differentiate into enterocyte-like cells by placing them in a culture medium without glucose for 18–21 days. The state of differentiation was evaluated by assaying the activity of enterocytic marker enzymes, and the acquisition of enterocyte morphology was assessed by electron microscopy. The α-
l-fucosidase activity was determined using a fluorometric method.
Intracellular levels of α-
l-fucosidase activity are lower in non-differentiated cells (3.0±1.01
U/mg) than in differentiated ones (9.2±4.09
U/mg) (
P<0.001). This variation is not due to a greater secretion of the enzyme to the culture medium, and properties such as pH optimum or the affinity towards substrate are not dependent on differentiation. The enzyme however, is more stable at acidic pH and at high temperatures, and
V
max is higher in differentiated cells. Moreover, in undifferentiated cells the enzyme is mainly in a monomeric form whereas multimeric forms of the enzyme appear only upon differentiation.
Most of these changes are very similar to those previously observed between normal colon tissue and colon tumours. Thus, we suggest that differentiation of HT-29 colon cancer cells could be used as a model to study the alterations of the enzyme α-
l-fucosidase during the progression of the tumoural process.</abstract><cop>Netherlands</cop><pub>Elsevier Ltd</pub><pmid>12127580</pmid><doi>10.1016/S1357-2725(02)00067-5</doi><tpages>13</tpages></addata></record> |
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| identifier | ISSN: 1357-2725 |
| ispartof | The international journal of biochemistry & cell biology, 2002-10, Vol.34 (10), p.1291-1303 |
| issn | 1357-2725 1878-5875 |
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| subjects | alpha-L-Fucosidase - metabolism Blotting, Western Cell Differentiation Chromatography, Gel Colonic Neoplasms - enzymology Colonic Neoplasms - pathology Colonic Neoplasms - ultrastructure Electrophoresis, Polyacrylamide Gel Enzymes HT-29 colon cancer cells HT29 Cells Humans Microscopy, Electron α- l-Fucosidase |
| title | Activity and properties of α- l-fucosidase are dependent on the state of enterocytic differentiation of HT-29 colon cancer cells |
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