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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Veröffentlicht in:International journal of oncology 2020-05, Vol.56 (5), p.1140-1151
Hauptverfasser: Cho, Woo-Cheol, Jang, Jee-Eun, Kim, Kyung-Hee, Yoo, Byong-Chul, Ku, Ja-Lok
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container_issue 5
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container_title International journal of oncology
container_volume 56
creator Cho, Woo-Cheol
Jang, Jee-Eun
Kim, Kyung-Hee
Yoo, Byong-Chul
Ku, Ja-Lok
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.
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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 &gt;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 &amp; 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 &gt;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. 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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 &gt;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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