SORBS1 serves a metastatic role via suppression of AHNAK in colorectal cancer cell lines
Cbl‑associated protein (CAP) is encoded by the sorbin and SH3 domain‑containing 1 (SORBS1) gene. CAP has been reported to be associated with the actin cytoskeleton, receptor tyrosine kinase signaling and cell adhesion through interactions with various proteins. It may be hypothesized that SORBS1 has...
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description | Cbl‑associated protein (CAP) is encoded by the sorbin and SH3 domain‑containing 1 (SORBS1) gene. CAP has been reported to be associated with the actin cytoskeleton, receptor tyrosine kinase signaling and cell adhesion through interactions with various proteins. It may be hypothesized that SORBS1 has numerous unknown functions, which may include providing a favorable condition for metastasis. Although CAP has been demonstrated to possess a number of functions, the role of this protein has only been reported in metabolic signaling pathways and its function in cancer remains to be elucidated. In the present study, SORBS1 expression was detected in colorectal cancer cell lines divided into the primary group and the metastatic group by reverse transcription‑quantitative PCR and western blot analysis. In addition, SORBS1 expression was manipulated by vector transfection and lentivirus transduction. The metastatic role of SORBS1, as determined by assessing its effects on cell proliferation and migration, was determined by colony formation assay, cell cycle analysis and Boyden chamber assay. To elucidate the SORBS1‑binding protein, immunoprecipitation was performed. Co‑localization of SORBS1 and AHNAK nucleoprotein (AHNAK) was identified by confocal microscopy. Notably, the protein expression levels of CAP were higher in SNU‑769A and SW480 cells than in SNU‑769B and SW620 cells. In addition, the number of colonies in the SORBS1‑overexpressing group was significantly increased compared with that of the control group, as determined using the colony formation assay; the SORBS1 overexpression group formed >8‑fold more colonies than the control group. The proliferative ability of the SORBS1 overexpression group was also significantly increased compared with the control group over the entire incubation period. Cell migration assays revealed that the number of migrated SORBS1‑knockdown cells was reduced compared with the control in both HCT‑116 and SNU‑C4 cell lines; migration area was decreased to 31 and 26% in HCT‑116 and SNU‑C4 cell lines, respectively. Consequently, it was confirmed that SORBS1 could form a complex with AHNAK, which functions as a tumor suppressor through inhibition of phosphorylated‑ERK and Rho‑associated coiled‑coil containing protein kinase 1. In conclusion, SORBS1 may serve a crucial role in cancer growth and migration via inhibition of AHNAK expression. |
doi_str_mv | 10.3892/ijo.2020.5006 |
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CAP has been reported to be associated with the actin cytoskeleton, receptor tyrosine kinase signaling and cell adhesion through interactions with various proteins. It may be hypothesized that SORBS1 has numerous unknown functions, which may include providing a favorable condition for metastasis. Although CAP has been demonstrated to possess a number of functions, the role of this protein has only been reported in metabolic signaling pathways and its function in cancer remains to be elucidated. In the present study, SORBS1 expression was detected in colorectal cancer cell lines divided into the primary group and the metastatic group by reverse transcription‑quantitative PCR and western blot analysis. In addition, SORBS1 expression was manipulated by vector transfection and lentivirus transduction. The metastatic role of SORBS1, as determined by assessing its effects on cell proliferation and migration, was determined by colony formation assay, cell cycle analysis and Boyden chamber assay. To elucidate the SORBS1‑binding protein, immunoprecipitation was performed. Co‑localization of SORBS1 and AHNAK nucleoprotein (AHNAK) was identified by confocal microscopy. Notably, the protein expression levels of CAP were higher in SNU‑769A and SW480 cells than in SNU‑769B and SW620 cells. In addition, the number of colonies in the SORBS1‑overexpressing group was significantly increased compared with that of the control group, as determined using the colony formation assay; the SORBS1 overexpression group formed >8‑fold more colonies than the control group. The proliferative ability of the SORBS1 overexpression group was also significantly increased compared with the control group over the entire incubation period. Cell migration assays revealed that the number of migrated SORBS1‑knockdown cells was reduced compared with the control in both HCT‑116 and SNU‑C4 cell lines; migration area was decreased to 31 and 26% in HCT‑116 and SNU‑C4 cell lines, respectively. Consequently, it was confirmed that SORBS1 could form a complex with AHNAK, which functions as a tumor suppressor through inhibition of phosphorylated‑ERK and Rho‑associated coiled‑coil containing protein kinase 1. In conclusion, SORBS1 may serve a crucial role in cancer growth and migration via inhibition of AHNAK expression.</description><identifier>ISSN: 1019-6439</identifier><identifier>EISSN: 1791-2423</identifier><identifier>DOI: 10.3892/ijo.2020.5006</identifier><identifier>PMID: 32319594</identifier><language>eng</language><publisher>Greece: Spandidos Publications</publisher><subject>Actin ; Adapter proteins ; Analysis ; Biotechnology ; Cancer diagnosis ; Cancer metastasis ; Cell adhesion & migration ; Cell cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colorectal cancer ; Colorectal Neoplasms - metabolism ; Deoxyribonucleic acid ; DNA ; DNA polymerase ; Gene Expression Regulation, Neoplastic ; Genes ; HCT116 Cells ; HT29 Cells ; Humans ; Instrument industry (Equipment) ; Insulin ; Kinases ; Laboratories ; Membrane Proteins - metabolism ; Metastasis ; Microfilament Proteins - genetics ; Microfilament Proteins - metabolism ; Microscopy ; Muscle proteins ; Neoplasm Metastasis ; Neoplasm Proteins - metabolism ; Pharmaceutical industry ; Phenols (Class of compounds) ; Polymerase chain reaction ; Protein binding ; Protein kinases ; Proteins ; Scientific equipment industry ; Signal Transduction ; Tumors ; Tyrosine ; Up-Regulation</subject><ispartof>International journal of oncology, 2020-05, Vol.56 (5), p.1140-1151</ispartof><rights>COPYRIGHT 2020 Spandidos Publications</rights><rights>Copyright Spandidos Publications UK Ltd. 2020</rights><rights>Copyright: © Cho et al. 2020</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c513t-35dc47669bc6a7074a7e52c75a187d44d930199dd66f68f52c93dbc6459bfd913</citedby><cites>FETCH-LOGICAL-c513t-35dc47669bc6a7074a7e52c75a187d44d930199dd66f68f52c93dbc6459bfd913</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32319594$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Cho, Woo-Cheol</creatorcontrib><creatorcontrib>Jang, Jee-Eun</creatorcontrib><creatorcontrib>Kim, Kyung-Hee</creatorcontrib><creatorcontrib>Yoo, Byong-Chul</creatorcontrib><creatorcontrib>Ku, Ja-Lok</creatorcontrib><title>SORBS1 serves a metastatic role via suppression of AHNAK in colorectal cancer cell lines</title><title>International journal of oncology</title><addtitle>Int J Oncol</addtitle><description>Cbl‑associated protein (CAP) is encoded by the sorbin and SH3 domain‑containing 1 (SORBS1) gene. CAP has been reported to be associated with the actin cytoskeleton, receptor tyrosine kinase signaling and cell adhesion through interactions with various proteins. It may be hypothesized that SORBS1 has numerous unknown functions, which may include providing a favorable condition for metastasis. Although CAP has been demonstrated to possess a number of functions, the role of this protein has only been reported in metabolic signaling pathways and its function in cancer remains to be elucidated. In the present study, SORBS1 expression was detected in colorectal cancer cell lines divided into the primary group and the metastatic group by reverse transcription‑quantitative PCR and western blot analysis. In addition, SORBS1 expression was manipulated by vector transfection and lentivirus transduction. The metastatic role of SORBS1, as determined by assessing its effects on cell proliferation and migration, was determined by colony formation assay, cell cycle analysis and Boyden chamber assay. To elucidate the SORBS1‑binding protein, immunoprecipitation was performed. Co‑localization of SORBS1 and AHNAK nucleoprotein (AHNAK) was identified by confocal microscopy. Notably, the protein expression levels of CAP were higher in SNU‑769A and SW480 cells than in SNU‑769B and SW620 cells. In addition, the number of colonies in the SORBS1‑overexpressing group was significantly increased compared with that of the control group, as determined using the colony formation assay; the SORBS1 overexpression group formed >8‑fold more colonies than the control group. The proliferative ability of the SORBS1 overexpression group was also significantly increased compared with the control group over the entire incubation period. Cell migration assays revealed that the number of migrated SORBS1‑knockdown cells was reduced compared with the control in both HCT‑116 and SNU‑C4 cell lines; migration area was decreased to 31 and 26% in HCT‑116 and SNU‑C4 cell lines, respectively. Consequently, it was confirmed that SORBS1 could form a complex with AHNAK, which functions as a tumor suppressor through inhibition of phosphorylated‑ERK and Rho‑associated coiled‑coil containing protein kinase 1. In conclusion, SORBS1 may serve a crucial role in cancer growth and migration via inhibition of AHNAK expression.</description><subject>Actin</subject><subject>Adapter proteins</subject><subject>Analysis</subject><subject>Biotechnology</subject><subject>Cancer diagnosis</subject><subject>Cancer metastasis</subject><subject>Cell adhesion & migration</subject><subject>Cell cycle</subject><subject>Cell Line, Tumor</subject><subject>Cell Movement</subject><subject>Cell Proliferation</subject><subject>Colorectal cancer</subject><subject>Colorectal Neoplasms - metabolism</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA polymerase</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>Genes</subject><subject>HCT116 Cells</subject><subject>HT29 Cells</subject><subject>Humans</subject><subject>Instrument industry (Equipment)</subject><subject>Insulin</subject><subject>Kinases</subject><subject>Laboratories</subject><subject>Membrane Proteins - metabolism</subject><subject>Metastasis</subject><subject>Microfilament Proteins - genetics</subject><subject>Microfilament Proteins - metabolism</subject><subject>Microscopy</subject><subject>Muscle proteins</subject><subject>Neoplasm Metastasis</subject><subject>Neoplasm Proteins - metabolism</subject><subject>Pharmaceutical industry</subject><subject>Phenols (Class of compounds)</subject><subject>Polymerase chain reaction</subject><subject>Protein binding</subject><subject>Protein kinases</subject><subject>Proteins</subject><subject>Scientific equipment industry</subject><subject>Signal Transduction</subject><subject>Tumors</subject><subject>Tyrosine</subject><subject>Up-Regulation</subject><issn>1019-6439</issn><issn>1791-2423</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNptks1rVDEUxR-i2FpdupWA4O6N-c5kI4xFrVgsWAV3IZN3XydDXjImeQP-92ZorR2QLBJyf_eQe3K67iXBC7bU9K3fpgXFFC8ExvJRd0qUJj3llD1uZ0x0LznTJ92zUrYYUyEwedqdMMqIFpqfdj-vr769vyaoQN5DQRZNUG2ptnqHcgqA9t6iMu92GUrxKaI0otXF19UX5CNyKaQMrtqAnI0OMnIQAgo-QnnePRltKPDibj_rfnz88P38or-8-vT5fHXZO0FY7ZkYHFdS6rWTVmHFrQJBnRKWLNXA-aBZG0IPg5SjXI6tpNnQWC70ehw0YWfdu1vd3byeYHAQa7bB7LKfbP5tkvXmuBL9xtykvVGECMUPAq_vBHL6NUOpZpvmHNubDW0OC0mFwv-oGxvA-DimJuYmX5xZSUqZopgsG7X4D9XWAJN3KcLo2_1Rw5sHDRuwoW5KCnNtVpdjsL8FXU6lZBjvJyTYHIJgWhDMIQjmEITGv3poyz399-fZH6G5q-I</recordid><startdate>20200501</startdate><enddate>20200501</enddate><creator>Cho, Woo-Cheol</creator><creator>Jang, Jee-Eun</creator><creator>Kim, Kyung-Hee</creator><creator>Yoo, Byong-Chul</creator><creator>Ku, Ja-Lok</creator><general>Spandidos Publications</general><general>Spandidos Publications UK Ltd</general><general>D.A. Spandidos</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>3V.</scope><scope>7RV</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AN0</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>5PM</scope></search><sort><creationdate>20200501</creationdate><title>SORBS1 serves a metastatic role via suppression of AHNAK in colorectal cancer cell lines</title><author>Cho, Woo-Cheol ; Jang, Jee-Eun ; Kim, Kyung-Hee ; Yoo, Byong-Chul ; Ku, Ja-Lok</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c513t-35dc47669bc6a7074a7e52c75a187d44d930199dd66f68f52c93dbc6459bfd913</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Actin</topic><topic>Adapter proteins</topic><topic>Analysis</topic><topic>Biotechnology</topic><topic>Cancer diagnosis</topic><topic>Cancer metastasis</topic><topic>Cell adhesion & migration</topic><topic>Cell cycle</topic><topic>Cell Line, Tumor</topic><topic>Cell Movement</topic><topic>Cell Proliferation</topic><topic>Colorectal cancer</topic><topic>Colorectal Neoplasms - metabolism</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA polymerase</topic><topic>Gene Expression Regulation, Neoplastic</topic><topic>Genes</topic><topic>HCT116 Cells</topic><topic>HT29 Cells</topic><topic>Humans</topic><topic>Instrument industry (Equipment)</topic><topic>Insulin</topic><topic>Kinases</topic><topic>Laboratories</topic><topic>Membrane Proteins - metabolism</topic><topic>Metastasis</topic><topic>Microfilament Proteins - genetics</topic><topic>Microfilament Proteins - metabolism</topic><topic>Microscopy</topic><topic>Muscle proteins</topic><topic>Neoplasm Metastasis</topic><topic>Neoplasm Proteins - metabolism</topic><topic>Pharmaceutical industry</topic><topic>Phenols (Class of compounds)</topic><topic>Polymerase chain reaction</topic><topic>Protein binding</topic><topic>Protein kinases</topic><topic>Proteins</topic><topic>Scientific equipment industry</topic><topic>Signal Transduction</topic><topic>Tumors</topic><topic>Tyrosine</topic><topic>Up-Regulation</topic><toplevel>online_resources</toplevel><creatorcontrib>Cho, Woo-Cheol</creatorcontrib><creatorcontrib>Jang, Jee-Eun</creatorcontrib><creatorcontrib>Kim, Kyung-Hee</creatorcontrib><creatorcontrib>Yoo, Byong-Chul</creatorcontrib><creatorcontrib>Ku, Ja-Lok</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Nursing & Allied Health Database</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>British Nursing Database</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Nursing & Allied Health Premium</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>International journal of oncology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cho, Woo-Cheol</au><au>Jang, Jee-Eun</au><au>Kim, Kyung-Hee</au><au>Yoo, Byong-Chul</au><au>Ku, Ja-Lok</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>SORBS1 serves a metastatic role via suppression of AHNAK in colorectal cancer cell lines</atitle><jtitle>International journal of oncology</jtitle><addtitle>Int J Oncol</addtitle><date>2020-05-01</date><risdate>2020</risdate><volume>56</volume><issue>5</issue><spage>1140</spage><epage>1151</epage><pages>1140-1151</pages><issn>1019-6439</issn><eissn>1791-2423</eissn><abstract>Cbl‑associated protein (CAP) is encoded by the sorbin and SH3 domain‑containing 1 (SORBS1) gene. CAP has been reported to be associated with the actin cytoskeleton, receptor tyrosine kinase signaling and cell adhesion through interactions with various proteins. It may be hypothesized that SORBS1 has numerous unknown functions, which may include providing a favorable condition for metastasis. Although CAP has been demonstrated to possess a number of functions, the role of this protein has only been reported in metabolic signaling pathways and its function in cancer remains to be elucidated. In the present study, SORBS1 expression was detected in colorectal cancer cell lines divided into the primary group and the metastatic group by reverse transcription‑quantitative PCR and western blot analysis. In addition, SORBS1 expression was manipulated by vector transfection and lentivirus transduction. The metastatic role of SORBS1, as determined by assessing its effects on cell proliferation and migration, was determined by colony formation assay, cell cycle analysis and Boyden chamber assay. To elucidate the SORBS1‑binding protein, immunoprecipitation was performed. Co‑localization of SORBS1 and AHNAK nucleoprotein (AHNAK) was identified by confocal microscopy. Notably, the protein expression levels of CAP were higher in SNU‑769A and SW480 cells than in SNU‑769B and SW620 cells. In addition, the number of colonies in the SORBS1‑overexpressing group was significantly increased compared with that of the control group, as determined using the colony formation assay; the SORBS1 overexpression group formed >8‑fold more colonies than the control group. The proliferative ability of the SORBS1 overexpression group was also significantly increased compared with the control group over the entire incubation period. Cell migration assays revealed that the number of migrated SORBS1‑knockdown cells was reduced compared with the control in both HCT‑116 and SNU‑C4 cell lines; migration area was decreased to 31 and 26% in HCT‑116 and SNU‑C4 cell lines, respectively. Consequently, it was confirmed that SORBS1 could form a complex with AHNAK, which functions as a tumor suppressor through inhibition of phosphorylated‑ERK and Rho‑associated coiled‑coil containing protein kinase 1. In conclusion, SORBS1 may serve a crucial role in cancer growth and migration via inhibition of AHNAK expression.</abstract><cop>Greece</cop><pub>Spandidos Publications</pub><pmid>32319594</pmid><doi>10.3892/ijo.2020.5006</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Actin Adapter proteins Analysis Biotechnology Cancer diagnosis Cancer metastasis Cell adhesion & migration Cell cycle Cell Line, Tumor Cell Movement Cell Proliferation Colorectal cancer Colorectal Neoplasms - metabolism Deoxyribonucleic acid DNA DNA polymerase Gene Expression Regulation, Neoplastic Genes HCT116 Cells HT29 Cells Humans Instrument industry (Equipment) Insulin Kinases Laboratories Membrane Proteins - metabolism Metastasis Microfilament Proteins - genetics Microfilament Proteins - metabolism Microscopy Muscle proteins Neoplasm Metastasis Neoplasm Proteins - metabolism Pharmaceutical industry Phenols (Class of compounds) Polymerase chain reaction Protein binding Protein kinases Proteins Scientific equipment industry Signal Transduction Tumors Tyrosine Up-Regulation |
title | SORBS1 serves a metastatic role via suppression of AHNAK in colorectal cancer cell lines |
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