Proteomic analysis of the TGF-β signaling pathway in pancreatic carcinoma cells using stable RNA interference to silence Smad4 expression
Smad4 is a tumor-suppressor gene that is lost or mutated in 50% of pancreatic carcinomas. Smad4 is also an intracellular transmitter of transforming growth factor-β (TGF-β) signals. Although its tumor-suppressor function is presumed to reside in its capacity to mediate TGF-β-induced growth inhibitio...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2004-05, Vol.318 (1), p.289-296 |
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| creator | Imamura, Takaaki Kanai, Fumihiko Kawakami, Takayuki Amarsanaa, Jazag Ijichi, Hideaki Hoshida, Yujin Tanaka, Yasuo Ikenoue, Tsuneo Tateishi, Keisuke Kawabe, Takao Arakawa, Yoshihiro Miyagishi, Makoto Taira, Kazunari Yokosuka, Osamu Omata, Masao |
| description | Smad4 is a tumor-suppressor gene that is lost or mutated in 50% of pancreatic carcinomas. Smad4 is also an intracellular transmitter of transforming growth factor-β (TGF-β) signals. Although its tumor-suppressor function is presumed to reside in its capacity to mediate TGF-β-induced growth inhibition, there seems to be a Smad4-independent TGF-β signaling pathway. Here, we succeeded in establishing Smad4 knockdown (S4KD) pancreatic cancer cell lines using stable RNA interference. Smad4 protein expression and TGF-β-Smad4 signaling were impaired in S4KD cells, and we compared the proteomic changes with TGF-β stimulation using two-dimensional gel electrophoresis (2-DE) and mass spectrometry. We identified five proteins that were up-regulated and seven proteins that were down-regulated; 10 of them were novel targets for TGF-β. These proteins function in processes such as cytoskeletal regulation, cell cycle, and oxidative stress. Introducing siRNA-mediated gene silencing into proteomics revealed a novel TGF-β signal pathway that did not involve Smad4. |
| doi_str_mv | 10.1016/j.bbrc.2004.04.029 |
| format | Article |
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Smad4 is also an intracellular transmitter of transforming growth factor-β (TGF-β) signals. Although its tumor-suppressor function is presumed to reside in its capacity to mediate TGF-β-induced growth inhibition, there seems to be a Smad4-independent TGF-β signaling pathway. Here, we succeeded in establishing Smad4 knockdown (S4KD) pancreatic cancer cell lines using stable RNA interference. Smad4 protein expression and TGF-β-Smad4 signaling were impaired in S4KD cells, and we compared the proteomic changes with TGF-β stimulation using two-dimensional gel electrophoresis (2-DE) and mass spectrometry. We identified five proteins that were up-regulated and seven proteins that were down-regulated; 10 of them were novel targets for TGF-β. These proteins function in processes such as cytoskeletal regulation, cell cycle, and oxidative stress. Introducing siRNA-mediated gene silencing into proteomics revealed a novel TGF-β signal pathway that did not involve Smad4.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2004.04.029</identifier><identifier>PMID: 15110786</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>2-DE ; Blotting, Western ; Cell Line, Tumor ; DNA-Binding Proteins - biosynthesis ; DNA-Binding Proteins - genetics ; Electrophoresis, Gel, Two-Dimensional ; Enzyme Inhibitors - pharmacology ; Flavonoids - pharmacology ; Gene Silencing ; Humans ; Mitogen-Activated Protein Kinases - antagonists & inhibitors ; Mitogen-Activated Protein Kinases - metabolism ; Pancreatic Neoplasms - genetics ; Pancreatic Neoplasms - metabolism ; Phosphorylation ; Protein Biosynthesis ; Proteins - analysis ; Proteome - genetics ; Proteome - metabolism ; Proteomics ; RNA Interference - physiology ; Signal Transduction ; Smad4 ; Smad4 Protein ; Stable RNAi ; TGF-β ; Trans-Activators - biosynthesis ; Trans-Activators - genetics ; Transforming Growth Factor beta - metabolism ; Transforming Growth Factor beta - pharmacology</subject><ispartof>Biochemical and biophysical research communications, 2004-05, Vol.318 (1), p.289-296</ispartof><rights>2004 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c385t-5e1bc1017a57fdb8557fa50f0300af2a0125878002efb109ba53827b9cb6bd963</citedby><cites>FETCH-LOGICAL-c385t-5e1bc1017a57fdb8557fa50f0300af2a0125878002efb109ba53827b9cb6bd963</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bbrc.2004.04.029$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15110786$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Imamura, Takaaki</creatorcontrib><creatorcontrib>Kanai, Fumihiko</creatorcontrib><creatorcontrib>Kawakami, Takayuki</creatorcontrib><creatorcontrib>Amarsanaa, Jazag</creatorcontrib><creatorcontrib>Ijichi, Hideaki</creatorcontrib><creatorcontrib>Hoshida, Yujin</creatorcontrib><creatorcontrib>Tanaka, Yasuo</creatorcontrib><creatorcontrib>Ikenoue, Tsuneo</creatorcontrib><creatorcontrib>Tateishi, Keisuke</creatorcontrib><creatorcontrib>Kawabe, Takao</creatorcontrib><creatorcontrib>Arakawa, Yoshihiro</creatorcontrib><creatorcontrib>Miyagishi, Makoto</creatorcontrib><creatorcontrib>Taira, Kazunari</creatorcontrib><creatorcontrib>Yokosuka, Osamu</creatorcontrib><creatorcontrib>Omata, Masao</creatorcontrib><title>Proteomic analysis of the TGF-β signaling pathway in pancreatic carcinoma cells using stable RNA interference to silence Smad4 expression</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>Smad4 is a tumor-suppressor gene that is lost or mutated in 50% of pancreatic carcinomas. 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Introducing siRNA-mediated gene silencing into proteomics revealed a novel TGF-β signal pathway that did not involve Smad4.</description><subject>2-DE</subject><subject>Blotting, Western</subject><subject>Cell Line, Tumor</subject><subject>DNA-Binding Proteins - biosynthesis</subject><subject>DNA-Binding Proteins - genetics</subject><subject>Electrophoresis, Gel, Two-Dimensional</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Flavonoids - pharmacology</subject><subject>Gene Silencing</subject><subject>Humans</subject><subject>Mitogen-Activated Protein Kinases - antagonists & inhibitors</subject><subject>Mitogen-Activated Protein Kinases - metabolism</subject><subject>Pancreatic Neoplasms - genetics</subject><subject>Pancreatic Neoplasms - metabolism</subject><subject>Phosphorylation</subject><subject>Protein Biosynthesis</subject><subject>Proteins - analysis</subject><subject>Proteome - genetics</subject><subject>Proteome - metabolism</subject><subject>Proteomics</subject><subject>RNA Interference - physiology</subject><subject>Signal Transduction</subject><subject>Smad4</subject><subject>Smad4 Protein</subject><subject>Stable RNAi</subject><subject>TGF-β</subject><subject>Trans-Activators - biosynthesis</subject><subject>Trans-Activators - genetics</subject><subject>Transforming Growth Factor beta - metabolism</subject><subject>Transforming Growth Factor beta - pharmacology</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkd9qFDEUxoModq2-gBeSK-9me05mM3_Am1JsFUotWsG7kGTOtFlmJmuSVfcVfBwfxGdqprvgnYUD50B-5-Pk-xh7jbBEwOpkvTQm2KUAWC3nEu0TtkBooRAIq6dsAQBVIVr8dsRexLgGQFxV7XN2hBIR6qZasN_XwSfyo7NcT3rYRRe573m6I35zcV78_cOju80PbrrlG53ufuodd1MeJxtIp7xmdbBu8qPmloYh8m2c2Zi0GYh_vjrNeKLQU6DJEk8-Cw4P45dRdytOvzaBYnR-esme9XqI9OrQj9nX8_c3Zx-Ky08XH89OLwtbNjIVktDY_P9ay7rvTCNz0xJ6KAF0LzSgkE3dAAjqTXbDaFk2ojatNZXp2qo8Zm_3upvgv28pJjW6ON-uJ_LbqGpsapBl-SiIDWBZIWRQ7EEbfIyBerUJbtRhpxDUHJVaqzkqNUel5hJtXnpzUN-akbp_K4dsMvBuD1A244ejoKJ1s3OdC2ST6rz7n_49R8-nBQ</recordid><startdate>20040521</startdate><enddate>20040521</enddate><creator>Imamura, Takaaki</creator><creator>Kanai, Fumihiko</creator><creator>Kawakami, Takayuki</creator><creator>Amarsanaa, Jazag</creator><creator>Ijichi, Hideaki</creator><creator>Hoshida, Yujin</creator><creator>Tanaka, Yasuo</creator><creator>Ikenoue, Tsuneo</creator><creator>Tateishi, Keisuke</creator><creator>Kawabe, Takao</creator><creator>Arakawa, Yoshihiro</creator><creator>Miyagishi, Makoto</creator><creator>Taira, Kazunari</creator><creator>Yokosuka, Osamu</creator><creator>Omata, Masao</creator><general>Elsevier Inc</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>7TM</scope><scope>7X8</scope></search><sort><creationdate>20040521</creationdate><title>Proteomic analysis of the TGF-β signaling pathway in pancreatic carcinoma cells using stable RNA interference to silence Smad4 expression</title><author>Imamura, Takaaki ; Kanai, Fumihiko ; Kawakami, Takayuki ; Amarsanaa, Jazag ; Ijichi, Hideaki ; Hoshida, Yujin ; Tanaka, Yasuo ; Ikenoue, Tsuneo ; Tateishi, Keisuke ; Kawabe, Takao ; Arakawa, Yoshihiro ; Miyagishi, Makoto ; Taira, Kazunari ; Yokosuka, Osamu ; Omata, Masao</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c385t-5e1bc1017a57fdb8557fa50f0300af2a0125878002efb109ba53827b9cb6bd963</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>2-DE</topic><topic>Blotting, Western</topic><topic>Cell Line, Tumor</topic><topic>DNA-Binding Proteins - biosynthesis</topic><topic>DNA-Binding Proteins - genetics</topic><topic>Electrophoresis, Gel, Two-Dimensional</topic><topic>Enzyme Inhibitors - pharmacology</topic><topic>Flavonoids - pharmacology</topic><topic>Gene Silencing</topic><topic>Humans</topic><topic>Mitogen-Activated Protein Kinases - antagonists & inhibitors</topic><topic>Mitogen-Activated Protein Kinases - metabolism</topic><topic>Pancreatic Neoplasms - genetics</topic><topic>Pancreatic Neoplasms - metabolism</topic><topic>Phosphorylation</topic><topic>Protein Biosynthesis</topic><topic>Proteins - analysis</topic><topic>Proteome - genetics</topic><topic>Proteome - metabolism</topic><topic>Proteomics</topic><topic>RNA Interference - physiology</topic><topic>Signal Transduction</topic><topic>Smad4</topic><topic>Smad4 Protein</topic><topic>Stable RNAi</topic><topic>TGF-β</topic><topic>Trans-Activators - biosynthesis</topic><topic>Trans-Activators - genetics</topic><topic>Transforming Growth Factor beta - metabolism</topic><topic>Transforming Growth Factor beta - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Imamura, Takaaki</creatorcontrib><creatorcontrib>Kanai, Fumihiko</creatorcontrib><creatorcontrib>Kawakami, Takayuki</creatorcontrib><creatorcontrib>Amarsanaa, Jazag</creatorcontrib><creatorcontrib>Ijichi, Hideaki</creatorcontrib><creatorcontrib>Hoshida, Yujin</creatorcontrib><creatorcontrib>Tanaka, Yasuo</creatorcontrib><creatorcontrib>Ikenoue, Tsuneo</creatorcontrib><creatorcontrib>Tateishi, Keisuke</creatorcontrib><creatorcontrib>Kawabe, Takao</creatorcontrib><creatorcontrib>Arakawa, Yoshihiro</creatorcontrib><creatorcontrib>Miyagishi, Makoto</creatorcontrib><creatorcontrib>Taira, Kazunari</creatorcontrib><creatorcontrib>Yokosuka, Osamu</creatorcontrib><creatorcontrib>Omata, Masao</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Imamura, Takaaki</au><au>Kanai, Fumihiko</au><au>Kawakami, Takayuki</au><au>Amarsanaa, Jazag</au><au>Ijichi, Hideaki</au><au>Hoshida, Yujin</au><au>Tanaka, Yasuo</au><au>Ikenoue, Tsuneo</au><au>Tateishi, Keisuke</au><au>Kawabe, Takao</au><au>Arakawa, Yoshihiro</au><au>Miyagishi, Makoto</au><au>Taira, Kazunari</au><au>Yokosuka, Osamu</au><au>Omata, Masao</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Proteomic analysis of the TGF-β signaling pathway in pancreatic carcinoma cells using stable RNA interference to silence Smad4 expression</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2004-05-21</date><risdate>2004</risdate><volume>318</volume><issue>1</issue><spage>289</spage><epage>296</epage><pages>289-296</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>Smad4 is a tumor-suppressor gene that is lost or mutated in 50% of pancreatic carcinomas. Smad4 is also an intracellular transmitter of transforming growth factor-β (TGF-β) signals. Although its tumor-suppressor function is presumed to reside in its capacity to mediate TGF-β-induced growth inhibition, there seems to be a Smad4-independent TGF-β signaling pathway. Here, we succeeded in establishing Smad4 knockdown (S4KD) pancreatic cancer cell lines using stable RNA interference. Smad4 protein expression and TGF-β-Smad4 signaling were impaired in S4KD cells, and we compared the proteomic changes with TGF-β stimulation using two-dimensional gel electrophoresis (2-DE) and mass spectrometry. We identified five proteins that were up-regulated and seven proteins that were down-regulated; 10 of them were novel targets for TGF-β. These proteins function in processes such as cytoskeletal regulation, cell cycle, and oxidative stress. Introducing siRNA-mediated gene silencing into proteomics revealed a novel TGF-β signal pathway that did not involve Smad4.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>15110786</pmid><doi>10.1016/j.bbrc.2004.04.029</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Biochemical and biophysical research communications, 2004-05, Vol.318 (1), p.289-296 |
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| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | 2-DE Blotting, Western Cell Line, Tumor DNA-Binding Proteins - biosynthesis DNA-Binding Proteins - genetics Electrophoresis, Gel, Two-Dimensional Enzyme Inhibitors - pharmacology Flavonoids - pharmacology Gene Silencing Humans Mitogen-Activated Protein Kinases - antagonists & inhibitors Mitogen-Activated Protein Kinases - metabolism Pancreatic Neoplasms - genetics Pancreatic Neoplasms - metabolism Phosphorylation Protein Biosynthesis Proteins - analysis Proteome - genetics Proteome - metabolism Proteomics RNA Interference - physiology Signal Transduction Smad4 Smad4 Protein Stable RNAi TGF-β Trans-Activators - biosynthesis Trans-Activators - genetics Transforming Growth Factor beta - metabolism Transforming Growth Factor beta - pharmacology |
| title | Proteomic analysis of the TGF-β signaling pathway in pancreatic carcinoma cells using stable RNA interference to silence Smad4 expression |
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