The Biological Activity of H. Pylori SlyD in Vitro
Aim To investigate the biological activity of the H. pylori SlyD in vitro. Methods Helicobacter pylori (H.pylori) slyD prokaryotic expression vector was carried out in Escherichia coli (E.coli), and recombination SlyD (rSlyD) was purified by immobilized metal affinity chromatography. The proliferati...
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| Veröffentlicht in: | Helicobacter (Cambridge, Mass.) Mass.), 2013-10, Vol.18 (5), p.347-355 |
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| creator | Dan, Kang Yuehua, Gong yanmei, Zhu aodi, Li nannan, Dong ying, Piao Yuan, Yuan |
| description | Aim
To investigate the biological activity of the H. pylori SlyD in vitro.
Methods
Helicobacter pylori (H.pylori) slyD prokaryotic expression vector was carried out in Escherichia coli (E.coli), and recombination SlyD (rSlyD) was purified by immobilized metal affinity chromatography. The proliferation, apoptosis, invasion, transformation effects of rSlyD on AGS cells was detected by CCK‐8, cell cycle, caspase‐3 activity, matrigel invasion assay, and double‐deck soft agar colony forming efficiency. In addition, the expressions of PCNA, KI‐67, caspase‐3, and MMP‐9 were detected by western blot and immunofluorescence assay, respectively.
Results
The CCK‐8 assay revealed that cell proliferation was increased in a time and dose‐dependent manner in AGS + rSlyD group compared with that of AGS or AGS + PBS group (p |
| doi_str_mv | 10.1111/hel.12057 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1434027394</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1434027394</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3117-f8293b3b8eaeb7f100b9e09bb672cbfece54c9e8e0ad04ce533ce32f1aa37c0a3</originalsourceid><addsrcrecordid>eNp1kM9SwjAQhzOOjiB68AWcHvVQSLotaY-ICA4ddQYUb5k0bCUaKDZF7dv4LD6ZVf7c3Mvuzn77HX6EnDLaZFW1ZmiazKMB3yN1FnjgBsDD_WqmIbg-hFGNHFn7QikNwI8OSc0DzvzqWicwnqFzqTOTPWsljdNRhX7XRelkqTNofn_dlybLtTMy5ZWjF86jLvLsmByk0lg82fQGebjujbsDN77r33Q7sauAMe6moRdBAkmIEhOeMkqTCGmUJG3uqSRFhYGvIgyRyin1qw1AIXgpkxK4ohIa5HztXebZ2wptIebaKjRGLjBbWcF88KnHIfIr9GKNqjyzNsdULHM9l3kpGBW_GYkqI_GXUcWebbSrZI7THbkNpQJaa-BDGyz_N4lBL94q3fWHtgV-7j5k_iraHHggJrd9QduTp9EwHgoGP3u4fu8</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1434027394</pqid></control><display><type>article</type><title>The Biological Activity of H. Pylori SlyD in Vitro</title><source>MEDLINE</source><source>Wiley Online Library All Journals</source><creator>Dan, Kang ; Yuehua, Gong ; yanmei, Zhu ; aodi, Li ; nannan, Dong ; ying, Piao ; Yuan, Yuan</creator><creatorcontrib>Dan, Kang ; Yuehua, Gong ; yanmei, Zhu ; aodi, Li ; nannan, Dong ; ying, Piao ; Yuan, Yuan</creatorcontrib><description>Aim
To investigate the biological activity of the H. pylori SlyD in vitro.
Methods
Helicobacter pylori (H.pylori) slyD prokaryotic expression vector was carried out in Escherichia coli (E.coli), and recombination SlyD (rSlyD) was purified by immobilized metal affinity chromatography. The proliferation, apoptosis, invasion, transformation effects of rSlyD on AGS cells was detected by CCK‐8, cell cycle, caspase‐3 activity, matrigel invasion assay, and double‐deck soft agar colony forming efficiency. In addition, the expressions of PCNA, KI‐67, caspase‐3, and MMP‐9 were detected by western blot and immunofluorescence assay, respectively.
Results
The CCK‐8 assay revealed that cell proliferation was increased in a time and dose‐dependent manner in AGS + rSlyD group compared with that of AGS or AGS + PBS group (p < .05). There are significant difference of PCNA and KI67 expressions among AGS, AGS + PBS, AGS + rSlyD groups (p < .05). Soft agar colony formation assay revealed the colony number (foci>100 μm) in AGS + rSlyD group was 26.3 ± 7.09, whereas 5.6 ± 1.15 in AGS and 5.0 ± 1.0 in AGS + PBS groups, respectively (p < .01). Colorimetric enzyme assay revealed the activity of caspase‐3 was decreased to 31.45 ± 0.49 after treatment with rSlyD, whereas 55.5 ± 0.43 in AGS and 55.1 ± 0.25 in AGS + PBS group, respectively (p < .001). Similar caspase‐3 expression also was confirmed by Western blot. The number of invasive cells in transwell chambers assay is 196.66 ± 40.41 in AGS + rSlyD group higher than 85 ± 22.9 in AGS or 81.66 ± 15.27 in AGS + PBS group, respectively (p < .001). The MMP‐9 expression in AGS + rSlyD group was also higher than that of AGS or AGS + PBS group.
Conclusion
These results suggest that the HpSlyD may play an important role in disturbing cell proliferation, apoptosis, and enhancing cell transformation and invasion in the AGS cell line. HpSlyD might contribute to gastric pathogenicity in H.pylori‐associated diseases.</description><identifier>ISSN: 1083-4389</identifier><identifier>EISSN: 1523-5378</identifier><identifier>DOI: 10.1111/hel.12057</identifier><identifier>PMID: 23714108</identifier><language>eng</language><publisher>England: Blackwell Publishing Ltd</publisher><subject>Apoptosis - drug effects ; Caspase 8 - metabolism ; Cell Line, Tumor ; Cell Proliferation - drug effects ; Cell Transformation, Neoplastic - drug effects ; Epithelial Cells - drug effects ; Escherichia coli ; Escherichia coli - genetics ; FK506-binding protein ; gastric cancer ; Gene Expression ; Gene Expression Profiling ; Helicobacter pylori ; Helicobacter pylori - enzymology ; Humans ; peptidyl-prolyl isomerase ; Peptidylprolyl Isomerase - genetics ; Peptidylprolyl Isomerase - isolation & purification ; Peptidylprolyl Isomerase - metabolism ; Recombinant Proteins - genetics ; Recombinant Proteins - isolation & purification ; Recombinant Proteins - metabolism ; Sincalide - metabolism ; slyD ; Virulence Factors - genetics ; Virulence Factors - isolation & purification ; Virulence Factors - metabolism</subject><ispartof>Helicobacter (Cambridge, Mass.), 2013-10, Vol.18 (5), p.347-355</ispartof><rights>2013 John Wiley & Sons Ltd</rights><rights>2013 John Wiley & Sons Ltd.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3117-f8293b3b8eaeb7f100b9e09bb672cbfece54c9e8e0ad04ce533ce32f1aa37c0a3</citedby><cites>FETCH-LOGICAL-c3117-f8293b3b8eaeb7f100b9e09bb672cbfece54c9e8e0ad04ce533ce32f1aa37c0a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fhel.12057$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fhel.12057$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23714108$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Dan, Kang</creatorcontrib><creatorcontrib>Yuehua, Gong</creatorcontrib><creatorcontrib>yanmei, Zhu</creatorcontrib><creatorcontrib>aodi, Li</creatorcontrib><creatorcontrib>nannan, Dong</creatorcontrib><creatorcontrib>ying, Piao</creatorcontrib><creatorcontrib>Yuan, Yuan</creatorcontrib><title>The Biological Activity of H. Pylori SlyD in Vitro</title><title>Helicobacter (Cambridge, Mass.)</title><addtitle>Helicobacter</addtitle><description>Aim
To investigate the biological activity of the H. pylori SlyD in vitro.
Methods
Helicobacter pylori (H.pylori) slyD prokaryotic expression vector was carried out in Escherichia coli (E.coli), and recombination SlyD (rSlyD) was purified by immobilized metal affinity chromatography. The proliferation, apoptosis, invasion, transformation effects of rSlyD on AGS cells was detected by CCK‐8, cell cycle, caspase‐3 activity, matrigel invasion assay, and double‐deck soft agar colony forming efficiency. In addition, the expressions of PCNA, KI‐67, caspase‐3, and MMP‐9 were detected by western blot and immunofluorescence assay, respectively.
Results
The CCK‐8 assay revealed that cell proliferation was increased in a time and dose‐dependent manner in AGS + rSlyD group compared with that of AGS or AGS + PBS group (p < .05). There are significant difference of PCNA and KI67 expressions among AGS, AGS + PBS, AGS + rSlyD groups (p < .05). Soft agar colony formation assay revealed the colony number (foci>100 μm) in AGS + rSlyD group was 26.3 ± 7.09, whereas 5.6 ± 1.15 in AGS and 5.0 ± 1.0 in AGS + PBS groups, respectively (p < .01). Colorimetric enzyme assay revealed the activity of caspase‐3 was decreased to 31.45 ± 0.49 after treatment with rSlyD, whereas 55.5 ± 0.43 in AGS and 55.1 ± 0.25 in AGS + PBS group, respectively (p < .001). Similar caspase‐3 expression also was confirmed by Western blot. The number of invasive cells in transwell chambers assay is 196.66 ± 40.41 in AGS + rSlyD group higher than 85 ± 22.9 in AGS or 81.66 ± 15.27 in AGS + PBS group, respectively (p < .001). The MMP‐9 expression in AGS + rSlyD group was also higher than that of AGS or AGS + PBS group.
Conclusion
These results suggest that the HpSlyD may play an important role in disturbing cell proliferation, apoptosis, and enhancing cell transformation and invasion in the AGS cell line. HpSlyD might contribute to gastric pathogenicity in H.pylori‐associated diseases.</description><subject>Apoptosis - drug effects</subject><subject>Caspase 8 - metabolism</subject><subject>Cell Line, Tumor</subject><subject>Cell Proliferation - drug effects</subject><subject>Cell Transformation, Neoplastic - drug effects</subject><subject>Epithelial Cells - drug effects</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>FK506-binding protein</subject><subject>gastric cancer</subject><subject>Gene Expression</subject><subject>Gene Expression Profiling</subject><subject>Helicobacter pylori</subject><subject>Helicobacter pylori - enzymology</subject><subject>Humans</subject><subject>peptidyl-prolyl isomerase</subject><subject>Peptidylprolyl Isomerase - genetics</subject><subject>Peptidylprolyl Isomerase - isolation & purification</subject><subject>Peptidylprolyl Isomerase - metabolism</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - metabolism</subject><subject>Sincalide - metabolism</subject><subject>slyD</subject><subject>Virulence Factors - genetics</subject><subject>Virulence Factors - isolation & purification</subject><subject>Virulence Factors - metabolism</subject><issn>1083-4389</issn><issn>1523-5378</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kM9SwjAQhzOOjiB68AWcHvVQSLotaY-ICA4ddQYUb5k0bCUaKDZF7dv4LD6ZVf7c3Mvuzn77HX6EnDLaZFW1ZmiazKMB3yN1FnjgBsDD_WqmIbg-hFGNHFn7QikNwI8OSc0DzvzqWicwnqFzqTOTPWsljdNRhX7XRelkqTNofn_dlybLtTMy5ZWjF86jLvLsmByk0lg82fQGebjujbsDN77r33Q7sauAMe6moRdBAkmIEhOeMkqTCGmUJG3uqSRFhYGvIgyRyin1qw1AIXgpkxK4ohIa5HztXebZ2wptIebaKjRGLjBbWcF88KnHIfIr9GKNqjyzNsdULHM9l3kpGBW_GYkqI_GXUcWebbSrZI7THbkNpQJaa-BDGyz_N4lBL94q3fWHtgV-7j5k_iraHHggJrd9QduTp9EwHgoGP3u4fu8</recordid><startdate>201310</startdate><enddate>201310</enddate><creator>Dan, Kang</creator><creator>Yuehua, Gong</creator><creator>yanmei, Zhu</creator><creator>aodi, Li</creator><creator>nannan, Dong</creator><creator>ying, Piao</creator><creator>Yuan, Yuan</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><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>7QL</scope><scope>C1K</scope></search><sort><creationdate>201310</creationdate><title>The Biological Activity of H. Pylori SlyD in Vitro</title><author>Dan, Kang ; Yuehua, Gong ; yanmei, Zhu ; aodi, Li ; nannan, Dong ; ying, Piao ; Yuan, Yuan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3117-f8293b3b8eaeb7f100b9e09bb672cbfece54c9e8e0ad04ce533ce32f1aa37c0a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Apoptosis - drug effects</topic><topic>Caspase 8 - metabolism</topic><topic>Cell Line, Tumor</topic><topic>Cell Proliferation - drug effects</topic><topic>Cell Transformation, Neoplastic - drug effects</topic><topic>Epithelial Cells - drug effects</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>FK506-binding protein</topic><topic>gastric cancer</topic><topic>Gene Expression</topic><topic>Gene Expression Profiling</topic><topic>Helicobacter pylori</topic><topic>Helicobacter pylori - enzymology</topic><topic>Humans</topic><topic>peptidyl-prolyl isomerase</topic><topic>Peptidylprolyl Isomerase - genetics</topic><topic>Peptidylprolyl Isomerase - isolation & purification</topic><topic>Peptidylprolyl Isomerase - metabolism</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Recombinant Proteins - metabolism</topic><topic>Sincalide - metabolism</topic><topic>slyD</topic><topic>Virulence Factors - genetics</topic><topic>Virulence Factors - isolation & purification</topic><topic>Virulence Factors - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Dan, Kang</creatorcontrib><creatorcontrib>Yuehua, Gong</creatorcontrib><creatorcontrib>yanmei, Zhu</creatorcontrib><creatorcontrib>aodi, Li</creatorcontrib><creatorcontrib>nannan, Dong</creatorcontrib><creatorcontrib>ying, Piao</creatorcontrib><creatorcontrib>Yuan, Yuan</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Helicobacter (Cambridge, Mass.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Dan, Kang</au><au>Yuehua, Gong</au><au>yanmei, Zhu</au><au>aodi, Li</au><au>nannan, Dong</au><au>ying, Piao</au><au>Yuan, Yuan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The Biological Activity of H. Pylori SlyD in Vitro</atitle><jtitle>Helicobacter (Cambridge, Mass.)</jtitle><addtitle>Helicobacter</addtitle><date>2013-10</date><risdate>2013</risdate><volume>18</volume><issue>5</issue><spage>347</spage><epage>355</epage><pages>347-355</pages><issn>1083-4389</issn><eissn>1523-5378</eissn><abstract>Aim
To investigate the biological activity of the H. pylori SlyD in vitro.
Methods
Helicobacter pylori (H.pylori) slyD prokaryotic expression vector was carried out in Escherichia coli (E.coli), and recombination SlyD (rSlyD) was purified by immobilized metal affinity chromatography. The proliferation, apoptosis, invasion, transformation effects of rSlyD on AGS cells was detected by CCK‐8, cell cycle, caspase‐3 activity, matrigel invasion assay, and double‐deck soft agar colony forming efficiency. In addition, the expressions of PCNA, KI‐67, caspase‐3, and MMP‐9 were detected by western blot and immunofluorescence assay, respectively.
Results
The CCK‐8 assay revealed that cell proliferation was increased in a time and dose‐dependent manner in AGS + rSlyD group compared with that of AGS or AGS + PBS group (p < .05). There are significant difference of PCNA and KI67 expressions among AGS, AGS + PBS, AGS + rSlyD groups (p < .05). Soft agar colony formation assay revealed the colony number (foci>100 μm) in AGS + rSlyD group was 26.3 ± 7.09, whereas 5.6 ± 1.15 in AGS and 5.0 ± 1.0 in AGS + PBS groups, respectively (p < .01). Colorimetric enzyme assay revealed the activity of caspase‐3 was decreased to 31.45 ± 0.49 after treatment with rSlyD, whereas 55.5 ± 0.43 in AGS and 55.1 ± 0.25 in AGS + PBS group, respectively (p < .001). Similar caspase‐3 expression also was confirmed by Western blot. The number of invasive cells in transwell chambers assay is 196.66 ± 40.41 in AGS + rSlyD group higher than 85 ± 22.9 in AGS or 81.66 ± 15.27 in AGS + PBS group, respectively (p < .001). The MMP‐9 expression in AGS + rSlyD group was also higher than that of AGS or AGS + PBS group.
Conclusion
These results suggest that the HpSlyD may play an important role in disturbing cell proliferation, apoptosis, and enhancing cell transformation and invasion in the AGS cell line. HpSlyD might contribute to gastric pathogenicity in H.pylori‐associated diseases.</abstract><cop>England</cop><pub>Blackwell Publishing Ltd</pub><pmid>23714108</pmid><doi>10.1111/hel.12057</doi><tpages>9</tpages></addata></record> |
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| subjects | Apoptosis - drug effects Caspase 8 - metabolism Cell Line, Tumor Cell Proliferation - drug effects Cell Transformation, Neoplastic - drug effects Epithelial Cells - drug effects Escherichia coli Escherichia coli - genetics FK506-binding protein gastric cancer Gene Expression Gene Expression Profiling Helicobacter pylori Helicobacter pylori - enzymology Humans peptidyl-prolyl isomerase Peptidylprolyl Isomerase - genetics Peptidylprolyl Isomerase - isolation & purification Peptidylprolyl Isomerase - metabolism Recombinant Proteins - genetics Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism Sincalide - metabolism slyD Virulence Factors - genetics Virulence Factors - isolation & purification Virulence Factors - metabolism |
| title | The Biological Activity of H. Pylori SlyD in Vitro |
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