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Knockdown of long non-coding RNA ANRIL inhibits tumorigenesis in human gastric cancer cells via microRNA-99a-mediated down-regulation of BMI1
Long non-coding RNA antisense non-coding RNA in the INK4 locus (ANRIL) has been reported to promote tumorigenesis via regulating microRNA (miR)-99a in gastric cancer cells. However, the role of each component involved in it is still not well understood. This study aimed to verify the role of ANRIL i...
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Published in: | Brazilian journal of medical and biological research 2018-01, Vol.51 (10), p.e6839 |
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description | Long non-coding RNA antisense non-coding RNA in the INK4 locus (ANRIL) has been reported to promote tumorigenesis via regulating microRNA (miR)-99a in gastric cancer cells. However, the role of each component involved in it is still not well understood. This study aimed to verify the role of ANRIL in gastric cancer as well as the underlying mechanisms. ANRIL levels in clinical gastric cancer tissues and cell lines were tested by qPCR. Effects of ANRIL silence on cell viability, migration and invasion, apoptosis, and miR-99a expression in MKN-45 and SGC-7901 cells were measured using CCK-8, Transwell assay, flow cytometry, and qPCR assays, respectively. Then, effects of miR-99a inhibition on ANRIL-silenced cells were evaluated. B-lymphoma Mo-MLV insertion region 1 (BMI1) expression, after abnormal expression of ANRIL and miR-99a, was determined. Finally, expression of key proteins in the apoptotic, Notch, and mTOR pathways was assessed. ANRIL level was elevated in gastric cancer tissues and cell lines. Knockdown of ANRIL suppressed cell viability, migration, and invasion, and increased apoptosis through up-regulating miR-99a. Furthermore, ANRIL silence down-regulated BMI1 via up-regulating miR-99a. BMI1 silence down-regulated Bcl-2 and key kinases in the Notch and mTOR pathways and up-regulated p16 and cleaved caspases. We verified the tumor suppressive effects of ANRIL knockdown in gastric cancer cells via crosstalk with miR-99a. Together, we provided a novel regulatory mechanism for ANRIL in gastric cancer, in which ANRIL silence down-regulated BMI1 via miR-99a, along with activation of the apoptotic pathway and inhibition of the Notch and mTOR pathways. |
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However, the role of each component involved in it is still not well understood. This study aimed to verify the role of ANRIL in gastric cancer as well as the underlying mechanisms. ANRIL levels in clinical gastric cancer tissues and cell lines were tested by qPCR. Effects of ANRIL silence on cell viability, migration and invasion, apoptosis, and miR-99a expression in MKN-45 and SGC-7901 cells were measured using CCK-8, Transwell assay, flow cytometry, and qPCR assays, respectively. Then, effects of miR-99a inhibition on ANRIL-silenced cells were evaluated. B-lymphoma Mo-MLV insertion region 1 (BMI1) expression, after abnormal expression of ANRIL and miR-99a, was determined. Finally, expression of key proteins in the apoptotic, Notch, and mTOR pathways was assessed. ANRIL level was elevated in gastric cancer tissues and cell lines. Knockdown of ANRIL suppressed cell viability, migration, and invasion, and increased apoptosis through up-regulating miR-99a. Furthermore, ANRIL silence down-regulated BMI1 via up-regulating miR-99a. BMI1 silence down-regulated Bcl-2 and key kinases in the Notch and mTOR pathways and up-regulated p16 and cleaved caspases. We verified the tumor suppressive effects of ANRIL knockdown in gastric cancer cells via crosstalk with miR-99a. Together, we provided a novel regulatory mechanism for ANRIL in gastric cancer, in which ANRIL silence down-regulated BMI1 via miR-99a, along with activation of the apoptotic pathway and inhibition of the Notch and mTOR pathways.</description><identifier>ISSN: 0100-879X</identifier><identifier>ISSN: 1414-431X</identifier><identifier>EISSN: 1414-431X</identifier><identifier>EISSN: 1678-4510</identifier><identifier>DOI: 10.1590/1414-431X20186839</identifier><identifier>PMID: 30156609</identifier><language>eng</language><publisher>Brazil: Associacao Brasileira de Divulgacao Cientifica (ABDC)</publisher><subject>Antisense RNA ; Apoptosis ; Apoptosis - genetics ; Apoptotic pathway ; Bcl-2 protein ; BIOLOGY ; BMI1 ; Carcinogenesis - genetics ; Cell Line, Tumor ; Cell migration ; Cholecystokinin ; Cyclin-dependent kinase inhibitors ; Development and progression ; Down-Regulation ; Flow cytometry ; Gastric cancer ; Gene Expression Regulation, Neoplastic ; Genetic aspects ; Humans ; INK4 protein ; Kinases ; lncRNA ANRIL ; Lymphocytes B ; Lymphoma ; MEDICINE, RESEARCH & EXPERIMENTAL ; MicroRNA ; MicroRNAs ; MicroRNAs - metabolism ; miR-99a ; miRNA ; Neoplasm Invasiveness ; Non-coding RNA ; Notch/mTOR pathway ; Physiological aspects ; RNA, Long Noncoding - genetics ; Stomach cancer ; Stomach Neoplasms - genetics ; Stomach Neoplasms - metabolism ; Stomach Neoplasms - pathology ; TOR protein ; TOR Serine-Threonine Kinases - metabolism ; Transfection ; Tumor cell lines ; Tumorigenesis ; Up-Regulation</subject><ispartof>Brazilian journal of medical and biological research, 2018-01, Vol.51 (10), p.e6839</ispartof><rights>COPYRIGHT 2018 Associacao Brasileira de Divulgacao Cientifica (ABDC)</rights><rights>Copyright Revista Brasileira de Pesquisas Medicas 2018</rights><rights>This work is licensed under a Creative Commons Attribution 4.0 International License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c657t-7a92cabc67c7835611be2adb89ebcf74345777baaec05a537fcebb19b4d5a4563</citedby><cites>FETCH-LOGICAL-c657t-7a92cabc67c7835611be2adb89ebcf74345777baaec05a537fcebb19b4d5a4563</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,24129,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30156609$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Liu, Pei</creatorcontrib><creatorcontrib>Zhang, Mingming</creatorcontrib><creatorcontrib>Niu, Qinghui</creatorcontrib><creatorcontrib>Zhang, Fengjuan</creatorcontrib><creatorcontrib>Yang, Yuling</creatorcontrib><creatorcontrib>Jiang, Xiangjun</creatorcontrib><title>Knockdown of long non-coding RNA ANRIL inhibits tumorigenesis in human gastric cancer cells via microRNA-99a-mediated down-regulation of BMI1</title><title>Brazilian journal of medical and biological research</title><addtitle>Braz J Med Biol Res</addtitle><description>Long non-coding RNA antisense non-coding RNA in the INK4 locus (ANRIL) has been reported to promote tumorigenesis via regulating microRNA (miR)-99a in gastric cancer cells. However, the role of each component involved in it is still not well understood. This study aimed to verify the role of ANRIL in gastric cancer as well as the underlying mechanisms. ANRIL levels in clinical gastric cancer tissues and cell lines were tested by qPCR. Effects of ANRIL silence on cell viability, migration and invasion, apoptosis, and miR-99a expression in MKN-45 and SGC-7901 cells were measured using CCK-8, Transwell assay, flow cytometry, and qPCR assays, respectively. Then, effects of miR-99a inhibition on ANRIL-silenced cells were evaluated. B-lymphoma Mo-MLV insertion region 1 (BMI1) expression, after abnormal expression of ANRIL and miR-99a, was determined. Finally, expression of key proteins in the apoptotic, Notch, and mTOR pathways was assessed. ANRIL level was elevated in gastric cancer tissues and cell lines. Knockdown of ANRIL suppressed cell viability, migration, and invasion, and increased apoptosis through up-regulating miR-99a. Furthermore, ANRIL silence down-regulated BMI1 via up-regulating miR-99a. BMI1 silence down-regulated Bcl-2 and key kinases in the Notch and mTOR pathways and up-regulated p16 and cleaved caspases. We verified the tumor suppressive effects of ANRIL knockdown in gastric cancer cells via crosstalk with miR-99a. Together, we provided a novel regulatory mechanism for ANRIL in gastric cancer, in which ANRIL silence down-regulated BMI1 via miR-99a, along with activation of the apoptotic pathway and inhibition of the Notch and mTOR pathways.</description><subject>Antisense RNA</subject><subject>Apoptosis</subject><subject>Apoptosis - genetics</subject><subject>Apoptotic pathway</subject><subject>Bcl-2 protein</subject><subject>BIOLOGY</subject><subject>BMI1</subject><subject>Carcinogenesis - genetics</subject><subject>Cell Line, Tumor</subject><subject>Cell migration</subject><subject>Cholecystokinin</subject><subject>Cyclin-dependent kinase inhibitors</subject><subject>Development and progression</subject><subject>Down-Regulation</subject><subject>Flow cytometry</subject><subject>Gastric cancer</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>Genetic aspects</subject><subject>Humans</subject><subject>INK4 protein</subject><subject>Kinases</subject><subject>lncRNA ANRIL</subject><subject>Lymphocytes B</subject><subject>Lymphoma</subject><subject>MEDICINE, RESEARCH & EXPERIMENTAL</subject><subject>MicroRNA</subject><subject>MicroRNAs</subject><subject>MicroRNAs - metabolism</subject><subject>miR-99a</subject><subject>miRNA</subject><subject>Neoplasm Invasiveness</subject><subject>Non-coding RNA</subject><subject>Notch/mTOR pathway</subject><subject>Physiological aspects</subject><subject>RNA, Long Noncoding - genetics</subject><subject>Stomach cancer</subject><subject>Stomach Neoplasms - genetics</subject><subject>Stomach Neoplasms - metabolism</subject><subject>Stomach Neoplasms - pathology</subject><subject>TOR protein</subject><subject>TOR Serine-Threonine Kinases - metabolism</subject><subject>Transfection</subject><subject>Tumor cell lines</subject><subject>Tumorigenesis</subject><subject>Up-Regulation</subject><issn>0100-879X</issn><issn>1414-431X</issn><issn>1414-431X</issn><issn>1678-4510</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNpVUk1v1DAQjRCILoUfwAVZQuKWYsd2HF-QthUfK5YiFZB6syaOk_WS2MVOCvwI_jNOt912T7bHb97MvHlZ9pLgE8IlfksYYTmj5LLApCorKh9li33scbbABOO8EvLyKHsW4xbjgmNGnmZHFBNellgusn-fndc_G__bId-i3rsOOe9y7RubrhfnS7Q8v1itkXUbW9sxonEafLCdcSbamMJoMw3gUAdxDFYjDU6bgLTp-4iuLaDB6uATTy4l5INpLIymQXPBPJhu6mG0_qb26ZcVeZ49aaGP5sXteZz9-PD--9mnfP314-psuc51ycWYC5CFhlqXQouK8pKQ2hTQ1JU0tW4Fo4wLIWoAozEHTkWrTV0TWbOGA-MlPc5WO97Gw1ZdBTtA-Ks8WHUT8KFTEEare6MYMKrLFlipNSuhqluWrlBR2rSGSJ24TnZcUVvTe7X1U3CpefVtll_N8s_rwfMLJ81TwrtdwtVUJ0G0cWOA_qCLwx9nN6rz1yrNiSkvEsHrW4Lgf00mjvc1C1KUsuCUyHtUB2kM61qfyPRgo1ZLnrYvqSiqhHrzALUx0I-b6Ptp3ko8BJIdMK0zxmDafcMEq9mNanaeSs77c-fGlPPq4aT7jDv70f91X9hM</recordid><startdate>20180101</startdate><enddate>20180101</enddate><creator>Liu, Pei</creator><creator>Zhang, Mingming</creator><creator>Niu, Qinghui</creator><creator>Zhang, Fengjuan</creator><creator>Yang, Yuling</creator><creator>Jiang, Xiangjun</creator><general>Associacao Brasileira de Divulgacao Cientifica (ABDC)</general><general>Revista Brasileira de Pesquisas Medicas</general><general>Associação Brasileira de Divulgação Científica</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>INF</scope><scope>7QG</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>8FD</scope><scope>FR3</scope><scope>K9.</scope><scope>P64</scope><scope>5PM</scope><scope>GPN</scope><scope>DOA</scope></search><sort><creationdate>20180101</creationdate><title>Knockdown of long non-coding RNA ANRIL inhibits tumorigenesis in human gastric cancer cells via microRNA-99a-mediated down-regulation of BMI1</title><author>Liu, Pei ; Zhang, Mingming ; Niu, Qinghui ; Zhang, Fengjuan ; Yang, Yuling ; Jiang, Xiangjun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c657t-7a92cabc67c7835611be2adb89ebcf74345777baaec05a537fcebb19b4d5a4563</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Antisense RNA</topic><topic>Apoptosis</topic><topic>Apoptosis - genetics</topic><topic>Apoptotic pathway</topic><topic>Bcl-2 protein</topic><topic>BIOLOGY</topic><topic>BMI1</topic><topic>Carcinogenesis - genetics</topic><topic>Cell Line, Tumor</topic><topic>Cell migration</topic><topic>Cholecystokinin</topic><topic>Cyclin-dependent kinase inhibitors</topic><topic>Development and progression</topic><topic>Down-Regulation</topic><topic>Flow cytometry</topic><topic>Gastric cancer</topic><topic>Gene Expression Regulation, Neoplastic</topic><topic>Genetic aspects</topic><topic>Humans</topic><topic>INK4 protein</topic><topic>Kinases</topic><topic>lncRNA ANRIL</topic><topic>Lymphocytes B</topic><topic>Lymphoma</topic><topic>MEDICINE, RESEARCH & EXPERIMENTAL</topic><topic>MicroRNA</topic><topic>MicroRNAs</topic><topic>MicroRNAs - metabolism</topic><topic>miR-99a</topic><topic>miRNA</topic><topic>Neoplasm Invasiveness</topic><topic>Non-coding RNA</topic><topic>Notch/mTOR pathway</topic><topic>Physiological aspects</topic><topic>RNA, Long Noncoding - genetics</topic><topic>Stomach cancer</topic><topic>Stomach Neoplasms - genetics</topic><topic>Stomach Neoplasms - metabolism</topic><topic>Stomach Neoplasms - pathology</topic><topic>TOR protein</topic><topic>TOR Serine-Threonine Kinases - metabolism</topic><topic>Transfection</topic><topic>Tumor cell lines</topic><topic>Tumorigenesis</topic><topic>Up-Regulation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Liu, Pei</creatorcontrib><creatorcontrib>Zhang, Mingming</creatorcontrib><creatorcontrib>Niu, Qinghui</creatorcontrib><creatorcontrib>Zhang, Fengjuan</creatorcontrib><creatorcontrib>Yang, Yuling</creatorcontrib><creatorcontrib>Jiang, Xiangjun</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale OneFile: Informe Academico</collection><collection>Animal Behavior Abstracts</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><collection>SciELO</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Brazilian journal of medical and biological research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Liu, Pei</au><au>Zhang, Mingming</au><au>Niu, Qinghui</au><au>Zhang, Fengjuan</au><au>Yang, Yuling</au><au>Jiang, Xiangjun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Knockdown of long non-coding RNA ANRIL inhibits tumorigenesis in human gastric cancer cells via microRNA-99a-mediated down-regulation of BMI1</atitle><jtitle>Brazilian journal of medical and biological research</jtitle><addtitle>Braz J Med Biol Res</addtitle><date>2018-01-01</date><risdate>2018</risdate><volume>51</volume><issue>10</issue><spage>e6839</spage><pages>e6839-</pages><issn>0100-879X</issn><issn>1414-431X</issn><eissn>1414-431X</eissn><eissn>1678-4510</eissn><abstract>Long non-coding RNA antisense non-coding RNA in the INK4 locus (ANRIL) has been reported to promote tumorigenesis via regulating microRNA (miR)-99a in gastric cancer cells. However, the role of each component involved in it is still not well understood. This study aimed to verify the role of ANRIL in gastric cancer as well as the underlying mechanisms. ANRIL levels in clinical gastric cancer tissues and cell lines were tested by qPCR. Effects of ANRIL silence on cell viability, migration and invasion, apoptosis, and miR-99a expression in MKN-45 and SGC-7901 cells were measured using CCK-8, Transwell assay, flow cytometry, and qPCR assays, respectively. Then, effects of miR-99a inhibition on ANRIL-silenced cells were evaluated. B-lymphoma Mo-MLV insertion region 1 (BMI1) expression, after abnormal expression of ANRIL and miR-99a, was determined. Finally, expression of key proteins in the apoptotic, Notch, and mTOR pathways was assessed. ANRIL level was elevated in gastric cancer tissues and cell lines. Knockdown of ANRIL suppressed cell viability, migration, and invasion, and increased apoptosis through up-regulating miR-99a. Furthermore, ANRIL silence down-regulated BMI1 via up-regulating miR-99a. BMI1 silence down-regulated Bcl-2 and key kinases in the Notch and mTOR pathways and up-regulated p16 and cleaved caspases. We verified the tumor suppressive effects of ANRIL knockdown in gastric cancer cells via crosstalk with miR-99a. Together, we provided a novel regulatory mechanism for ANRIL in gastric cancer, in which ANRIL silence down-regulated BMI1 via miR-99a, along with activation of the apoptotic pathway and inhibition of the Notch and mTOR pathways.</abstract><cop>Brazil</cop><pub>Associacao Brasileira de Divulgacao Cientifica (ABDC)</pub><pmid>30156609</pmid><doi>10.1590/1414-431X20186839</doi><oa>free_for_read</oa></addata></record> |
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subjects | Antisense RNA Apoptosis Apoptosis - genetics Apoptotic pathway Bcl-2 protein BIOLOGY BMI1 Carcinogenesis - genetics Cell Line, Tumor Cell migration Cholecystokinin Cyclin-dependent kinase inhibitors Development and progression Down-Regulation Flow cytometry Gastric cancer Gene Expression Regulation, Neoplastic Genetic aspects Humans INK4 protein Kinases lncRNA ANRIL Lymphocytes B Lymphoma MEDICINE, RESEARCH & EXPERIMENTAL MicroRNA MicroRNAs MicroRNAs - metabolism miR-99a miRNA Neoplasm Invasiveness Non-coding RNA Notch/mTOR pathway Physiological aspects RNA, Long Noncoding - genetics Stomach cancer Stomach Neoplasms - genetics Stomach Neoplasms - metabolism Stomach Neoplasms - pathology TOR protein TOR Serine-Threonine Kinases - metabolism Transfection Tumor cell lines Tumorigenesis Up-Regulation |
title | Knockdown of long non-coding RNA ANRIL inhibits tumorigenesis in human gastric cancer cells via microRNA-99a-mediated down-regulation of BMI1 |
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