Knockdown of FAM3B triggers cell apoptosis through p53-dependent pathway
FAM3B, also named PANDER, is a cytokine-like protein identified in 2002. Previous studies showed that FAM3B regulates glucose and lipid metabolism through interaction with liver and endocrine pancreas. FAM3B is also expressed by other tissues but its basic function is unclear. In this study, we foun...
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description | FAM3B, also named PANDER, is a cytokine-like protein identified in 2002. Previous studies showed that FAM3B regulates glucose and lipid metabolism through interaction with liver and endocrine pancreas. FAM3B is also expressed by other tissues but its basic function is unclear. In this study, we found that FAM3B was expressed in mouse colon, intestine, liver and lung tissues and multiple types of cell lines, including murine pancreatic β-cell (Min6), microglia (N9) and muscle cell (C2C12); human colon cancer cells (HCT8, HCT116, HT29), hepatocyte (HL-7702), hepatocellular carcinoma cell (SMMC-7721) and lung carcinoma cell (A549). Inhibition of FAM3B expression by RNA interference induced apoptotic cell death of HCT8, HCT116, A549, N9, C2C12 and Min6 cells and decreased cell viability of HL-7702 and murine primary hepatocytes. Further studies with HCT8 cells showed that knockdown of FAM3B increased the protein levels of membrane-bound Fas and Bax, reduced the expression of Bcl-2, promoted the cleavage of caspases-8, -3, -9 and PARP, and the nuclear translocation of cleaved PARP. These results suggest that FAM3B silencing activates both extrinsic and intrinsic apoptotic pathways. Mechanistic studies showed that neutralizing antibody against Fas or silencing Fas-associated death domain had no effect on, while caspase inhibitors could significantly reverse FAM3B knockdown induced apoptosis, suggesting Fas and death receptor mediated extrinsic apoptotic pathway is not involved in FAM3B silencing induced apoptosis. Further studies showed that p53 was significantly upregulated after FAM3B knockdown. Silencing p53 could almost completely reverse FAM3B knockdown induced upregulation of Bax, downregulation of Bcl-2, cleavage of caspases-8, -9, -3, and apoptotic cell death, suggesting p53-dependent pathway plays critical roles in FAM3B silencing induced apoptosis. Studies with HCT116 cells confirmed that inhibition of FAM3B expression induced apoptosis through p53-dependent pathway. Furthermore, knockdown of FAM3B reduced the protein level of Mdm2 and promoted p53 phosphorylation. Taken together, our studies demonstrated that silencing FAM3B promoted p53 phosphorylation and induced p53 accumulation by decreasing Mdm2 expression, which resulted in apoptotic cell death. |
doi_str_mv | 10.1016/j.biocel.2012.12.003 |
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Previous studies showed that FAM3B regulates glucose and lipid metabolism through interaction with liver and endocrine pancreas. FAM3B is also expressed by other tissues but its basic function is unclear. In this study, we found that FAM3B was expressed in mouse colon, intestine, liver and lung tissues and multiple types of cell lines, including murine pancreatic β-cell (Min6), microglia (N9) and muscle cell (C2C12); human colon cancer cells (HCT8, HCT116, HT29), hepatocyte (HL-7702), hepatocellular carcinoma cell (SMMC-7721) and lung carcinoma cell (A549). Inhibition of FAM3B expression by RNA interference induced apoptotic cell death of HCT8, HCT116, A549, N9, C2C12 and Min6 cells and decreased cell viability of HL-7702 and murine primary hepatocytes. Further studies with HCT8 cells showed that knockdown of FAM3B increased the protein levels of membrane-bound Fas and Bax, reduced the expression of Bcl-2, promoted the cleavage of caspases-8, -3, -9 and PARP, and the nuclear translocation of cleaved PARP. These results suggest that FAM3B silencing activates both extrinsic and intrinsic apoptotic pathways. Mechanistic studies showed that neutralizing antibody against Fas or silencing Fas-associated death domain had no effect on, while caspase inhibitors could significantly reverse FAM3B knockdown induced apoptosis, suggesting Fas and death receptor mediated extrinsic apoptotic pathway is not involved in FAM3B silencing induced apoptosis. Further studies showed that p53 was significantly upregulated after FAM3B knockdown. Silencing p53 could almost completely reverse FAM3B knockdown induced upregulation of Bax, downregulation of Bcl-2, cleavage of caspases-8, -9, -3, and apoptotic cell death, suggesting p53-dependent pathway plays critical roles in FAM3B silencing induced apoptosis. Studies with HCT116 cells confirmed that inhibition of FAM3B expression induced apoptosis through p53-dependent pathway. Furthermore, knockdown of FAM3B reduced the protein level of Mdm2 and promoted p53 phosphorylation. Taken together, our studies demonstrated that silencing FAM3B promoted p53 phosphorylation and induced p53 accumulation by decreasing Mdm2 expression, which resulted in apoptotic cell death.</description><identifier>ISSN: 1357-2725</identifier><identifier>EISSN: 1878-5875</identifier><identifier>DOI: 10.1016/j.biocel.2012.12.003</identifier><identifier>PMID: 23246487</identifier><language>eng</language><publisher>Netherlands: Elsevier Ltd</publisher><subject>Animals ; Apoptosis ; Apoptosis - genetics ; Caspase ; Cell viability ; Cytokines - biosynthesis ; Cytokines - genetics ; FAM3B ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; HCT116 Cells ; Humans ; Mdm2 ; Mice ; Neoplasm Proteins - biosynthesis ; Neoplasm Proteins - genetics ; Neoplasms - genetics ; Neoplasms - metabolism ; p53 ; PANDER ; Phosphorylation ; Proto-Oncogene Proteins c-mdm2 - metabolism ; Signal Transduction ; Tumor Suppressor Protein p53 - biosynthesis ; Tumor Suppressor Protein p53 - genetics</subject><ispartof>The international journal of biochemistry & cell biology, 2013-03, Vol.45 (3), p.684-691</ispartof><rights>2012 Elsevier Ltd</rights><rights>Copyright © 2012 Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c362t-45f5b6378a28fd72ea759126fd1f46ae9bc5268ca09f56d010f7e09d0f4da4eb3</citedby><cites>FETCH-LOGICAL-c362t-45f5b6378a28fd72ea759126fd1f46ae9bc5268ca09f56d010f7e09d0f4da4eb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.biocel.2012.12.003$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23246487$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mou, Haiwei</creatorcontrib><creatorcontrib>Li, Zongmeng</creatorcontrib><creatorcontrib>Yao, Pengle</creatorcontrib><creatorcontrib>Zhuo, Shu</creatorcontrib><creatorcontrib>Luan, Wei</creatorcontrib><creatorcontrib>Deng, Bo</creatorcontrib><creatorcontrib>Qian, Lihua</creatorcontrib><creatorcontrib>Yang, Mengmei</creatorcontrib><creatorcontrib>Mei, Hong</creatorcontrib><creatorcontrib>Le, Yingying</creatorcontrib><title>Knockdown of FAM3B triggers cell apoptosis through p53-dependent pathway</title><title>The international journal of biochemistry & cell biology</title><addtitle>Int J Biochem Cell Biol</addtitle><description>FAM3B, also named PANDER, is a cytokine-like protein identified in 2002. Previous studies showed that FAM3B regulates glucose and lipid metabolism through interaction with liver and endocrine pancreas. FAM3B is also expressed by other tissues but its basic function is unclear. In this study, we found that FAM3B was expressed in mouse colon, intestine, liver and lung tissues and multiple types of cell lines, including murine pancreatic β-cell (Min6), microglia (N9) and muscle cell (C2C12); human colon cancer cells (HCT8, HCT116, HT29), hepatocyte (HL-7702), hepatocellular carcinoma cell (SMMC-7721) and lung carcinoma cell (A549). Inhibition of FAM3B expression by RNA interference induced apoptotic cell death of HCT8, HCT116, A549, N9, C2C12 and Min6 cells and decreased cell viability of HL-7702 and murine primary hepatocytes. Further studies with HCT8 cells showed that knockdown of FAM3B increased the protein levels of membrane-bound Fas and Bax, reduced the expression of Bcl-2, promoted the cleavage of caspases-8, -3, -9 and PARP, and the nuclear translocation of cleaved PARP. These results suggest that FAM3B silencing activates both extrinsic and intrinsic apoptotic pathways. Mechanistic studies showed that neutralizing antibody against Fas or silencing Fas-associated death domain had no effect on, while caspase inhibitors could significantly reverse FAM3B knockdown induced apoptosis, suggesting Fas and death receptor mediated extrinsic apoptotic pathway is not involved in FAM3B silencing induced apoptosis. Further studies showed that p53 was significantly upregulated after FAM3B knockdown. Silencing p53 could almost completely reverse FAM3B knockdown induced upregulation of Bax, downregulation of Bcl-2, cleavage of caspases-8, -9, -3, and apoptotic cell death, suggesting p53-dependent pathway plays critical roles in FAM3B silencing induced apoptosis. Studies with HCT116 cells confirmed that inhibition of FAM3B expression induced apoptosis through p53-dependent pathway. Furthermore, knockdown of FAM3B reduced the protein level of Mdm2 and promoted p53 phosphorylation. Taken together, our studies demonstrated that silencing FAM3B promoted p53 phosphorylation and induced p53 accumulation by decreasing Mdm2 expression, which resulted in apoptotic cell death.</description><subject>Animals</subject><subject>Apoptosis</subject><subject>Apoptosis - genetics</subject><subject>Caspase</subject><subject>Cell viability</subject><subject>Cytokines - biosynthesis</subject><subject>Cytokines - genetics</subject><subject>FAM3B</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>Gene Knockdown Techniques</subject><subject>HCT116 Cells</subject><subject>Humans</subject><subject>Mdm2</subject><subject>Mice</subject><subject>Neoplasm Proteins - biosynthesis</subject><subject>Neoplasm Proteins - genetics</subject><subject>Neoplasms - genetics</subject><subject>Neoplasms - metabolism</subject><subject>p53</subject><subject>PANDER</subject><subject>Phosphorylation</subject><subject>Proto-Oncogene Proteins c-mdm2 - metabolism</subject><subject>Signal Transduction</subject><subject>Tumor Suppressor Protein p53 - biosynthesis</subject><subject>Tumor Suppressor Protein p53 - genetics</subject><issn>1357-2725</issn><issn>1878-5875</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMtOwzAQRS0E4lH4A4SyZJPiR2wnGySoeIkiNrC2HHvcurRxsFMQf4-rAkukkWYW996ZOQidEjwmmIiLxbj1wcByTDGh41wYsx10SGpZl7yWfDfPjMuSSsoP0FFKC4wx4ZTtowPKaCWqWh6i-8cumDcbPrsiuOL26oldF0P0sxnEVOT0ZaH70A8h-VQM8xjWs3nRc1Za6KGz0A1Fr4f5p_46RntOLxOc_PQRer29eZncl9Pnu4fJ1bQ0TNChrLjjrWCy1rR2VlLQkjeECmeJq4SGpjWcitpo3DguLCbYScCNxa6yuoKWjdD5NreP4X0NaVArnzaH6g7COilCa14JwUiTpdVWamJIKYJTffQrHb8UwWrDUC3UlqHaMMxWlRlm29nPhnW7Avtn-oWWBZdbAeQ_PzxElYyHzoD1EcygbPD_b_gGsGODzw</recordid><startdate>201303</startdate><enddate>201303</enddate><creator>Mou, Haiwei</creator><creator>Li, Zongmeng</creator><creator>Yao, Pengle</creator><creator>Zhuo, Shu</creator><creator>Luan, Wei</creator><creator>Deng, Bo</creator><creator>Qian, Lihua</creator><creator>Yang, Mengmei</creator><creator>Mei, Hong</creator><creator>Le, Yingying</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>201303</creationdate><title>Knockdown of FAM3B triggers cell apoptosis through p53-dependent pathway</title><author>Mou, Haiwei ; Li, Zongmeng ; Yao, Pengle ; Zhuo, Shu ; Luan, Wei ; Deng, Bo ; Qian, Lihua ; Yang, Mengmei ; Mei, Hong ; Le, Yingying</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c362t-45f5b6378a28fd72ea759126fd1f46ae9bc5268ca09f56d010f7e09d0f4da4eb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animals</topic><topic>Apoptosis</topic><topic>Apoptosis - genetics</topic><topic>Caspase</topic><topic>Cell viability</topic><topic>Cytokines - biosynthesis</topic><topic>Cytokines - genetics</topic><topic>FAM3B</topic><topic>Gene Expression Regulation, Neoplastic</topic><topic>Gene Knockdown Techniques</topic><topic>HCT116 Cells</topic><topic>Humans</topic><topic>Mdm2</topic><topic>Mice</topic><topic>Neoplasm Proteins - biosynthesis</topic><topic>Neoplasm Proteins - genetics</topic><topic>Neoplasms - genetics</topic><topic>Neoplasms - metabolism</topic><topic>p53</topic><topic>PANDER</topic><topic>Phosphorylation</topic><topic>Proto-Oncogene Proteins c-mdm2 - metabolism</topic><topic>Signal Transduction</topic><topic>Tumor Suppressor Protein p53 - biosynthesis</topic><topic>Tumor Suppressor Protein p53 - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mou, Haiwei</creatorcontrib><creatorcontrib>Li, Zongmeng</creatorcontrib><creatorcontrib>Yao, Pengle</creatorcontrib><creatorcontrib>Zhuo, Shu</creatorcontrib><creatorcontrib>Luan, Wei</creatorcontrib><creatorcontrib>Deng, Bo</creatorcontrib><creatorcontrib>Qian, Lihua</creatorcontrib><creatorcontrib>Yang, Mengmei</creatorcontrib><creatorcontrib>Mei, Hong</creatorcontrib><creatorcontrib>Le, Yingying</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>Mou, Haiwei</au><au>Li, Zongmeng</au><au>Yao, Pengle</au><au>Zhuo, Shu</au><au>Luan, Wei</au><au>Deng, Bo</au><au>Qian, Lihua</au><au>Yang, Mengmei</au><au>Mei, Hong</au><au>Le, Yingying</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Knockdown of FAM3B triggers cell apoptosis through p53-dependent pathway</atitle><jtitle>The international journal of biochemistry & cell biology</jtitle><addtitle>Int J Biochem Cell Biol</addtitle><date>2013-03</date><risdate>2013</risdate><volume>45</volume><issue>3</issue><spage>684</spage><epage>691</epage><pages>684-691</pages><issn>1357-2725</issn><eissn>1878-5875</eissn><abstract>FAM3B, also named PANDER, is a cytokine-like protein identified in 2002. Previous studies showed that FAM3B regulates glucose and lipid metabolism through interaction with liver and endocrine pancreas. FAM3B is also expressed by other tissues but its basic function is unclear. In this study, we found that FAM3B was expressed in mouse colon, intestine, liver and lung tissues and multiple types of cell lines, including murine pancreatic β-cell (Min6), microglia (N9) and muscle cell (C2C12); human colon cancer cells (HCT8, HCT116, HT29), hepatocyte (HL-7702), hepatocellular carcinoma cell (SMMC-7721) and lung carcinoma cell (A549). Inhibition of FAM3B expression by RNA interference induced apoptotic cell death of HCT8, HCT116, A549, N9, C2C12 and Min6 cells and decreased cell viability of HL-7702 and murine primary hepatocytes. Further studies with HCT8 cells showed that knockdown of FAM3B increased the protein levels of membrane-bound Fas and Bax, reduced the expression of Bcl-2, promoted the cleavage of caspases-8, -3, -9 and PARP, and the nuclear translocation of cleaved PARP. These results suggest that FAM3B silencing activates both extrinsic and intrinsic apoptotic pathways. Mechanistic studies showed that neutralizing antibody against Fas or silencing Fas-associated death domain had no effect on, while caspase inhibitors could significantly reverse FAM3B knockdown induced apoptosis, suggesting Fas and death receptor mediated extrinsic apoptotic pathway is not involved in FAM3B silencing induced apoptosis. Further studies showed that p53 was significantly upregulated after FAM3B knockdown. Silencing p53 could almost completely reverse FAM3B knockdown induced upregulation of Bax, downregulation of Bcl-2, cleavage of caspases-8, -9, -3, and apoptotic cell death, suggesting p53-dependent pathway plays critical roles in FAM3B silencing induced apoptosis. Studies with HCT116 cells confirmed that inhibition of FAM3B expression induced apoptosis through p53-dependent pathway. Furthermore, knockdown of FAM3B reduced the protein level of Mdm2 and promoted p53 phosphorylation. Taken together, our studies demonstrated that silencing FAM3B promoted p53 phosphorylation and induced p53 accumulation by decreasing Mdm2 expression, which resulted in apoptotic cell death.</abstract><cop>Netherlands</cop><pub>Elsevier Ltd</pub><pmid>23246487</pmid><doi>10.1016/j.biocel.2012.12.003</doi><tpages>8</tpages></addata></record> |
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subjects | Animals Apoptosis Apoptosis - genetics Caspase Cell viability Cytokines - biosynthesis Cytokines - genetics FAM3B Gene Expression Regulation, Neoplastic Gene Knockdown Techniques HCT116 Cells Humans Mdm2 Mice Neoplasm Proteins - biosynthesis Neoplasm Proteins - genetics Neoplasms - genetics Neoplasms - metabolism p53 PANDER Phosphorylation Proto-Oncogene Proteins c-mdm2 - metabolism Signal Transduction Tumor Suppressor Protein p53 - biosynthesis Tumor Suppressor Protein p53 - genetics |
title | Knockdown of FAM3B triggers cell apoptosis through p53-dependent pathway |
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