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Identification of LINC02310 as an enhancer in lung adenocarcinoma and investigation of its regulatory network via comprehensive analyses
Lung adenocarcinoma (LADC) is a major subtype of non-small cell lung cancer and has one of the highest mortality rates. An increasing number of long non-coding RNAs (LncRNAs) were reported to be associated with the occurrence and progression of LADC. Thus, it is necessary and reasonable to find new...
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Published in: | BMC medical genomics 2020-12, Vol.13 (1), p.185-185, Article 185 |
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description | Lung adenocarcinoma (LADC) is a major subtype of non-small cell lung cancer and has one of the highest mortality rates. An increasing number of long non-coding RNAs (LncRNAs) were reported to be associated with the occurrence and progression of LADC. Thus, it is necessary and reasonable to find new prognostic biomarkers for LADC among LncRNAs.
Differential expression analysis, survival analysis, PCR experiments and clinical feature analysis were performed to screen out the LncRNA which was significantly related to LADC. Its role in LADC was verified by CCK-8 assay and colony. Furthermore, competing endogenous RNA (ceRNA) regulatory network construction, enrichment analysis and protein-protein interaction (PPI) network construction were performed to investigate the downstream regulatory network of the selected LncRNA.
A total of 2431 differentially expressed LncRNAs (DELncRNAs) and 2227 differentially expressed mRNAs (DEmRNAs) were from The Cancer Genome Atlas database. Survival analysis results indicated that lnc-YARS2-5, lnc-NPR3-2 and LINC02310 were significantly related to overall survival. Their overexpression indicated poor prognostic. PCR experiments and clinical feature analysis suggested that LINC02310 was significantly correlated with TNM-stage and T-stage. CCK-8 assay and colony formation assay demonstrated that LINC02310 acted as an enhancer in LADC. In addition, 3 targeted miRNAs of LINC02310 and 414 downstream DEmRNAs were predicted. The downstream DEmRNAs were then enriched in 405 Gene Ontology terms and 11 Kyoto Encyclopedia of Genes and Genomes pathways, which revealed their potential functions and mechanisms. The PPI network showed the interactions among the downstream DEmRNAs.
This study verified LINC02310 as an enhancer in LADC and performed comprehensive analyses on its downstream regulatory network, which might benefit LADC prognoses and therapies. |
doi_str_mv | 10.1186/s12920-020-00834-6 |
format | article |
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Differential expression analysis, survival analysis, PCR experiments and clinical feature analysis were performed to screen out the LncRNA which was significantly related to LADC. Its role in LADC was verified by CCK-8 assay and colony. Furthermore, competing endogenous RNA (ceRNA) regulatory network construction, enrichment analysis and protein-protein interaction (PPI) network construction were performed to investigate the downstream regulatory network of the selected LncRNA.
A total of 2431 differentially expressed LncRNAs (DELncRNAs) and 2227 differentially expressed mRNAs (DEmRNAs) were from The Cancer Genome Atlas database. Survival analysis results indicated that lnc-YARS2-5, lnc-NPR3-2 and LINC02310 were significantly related to overall survival. Their overexpression indicated poor prognostic. PCR experiments and clinical feature analysis suggested that LINC02310 was significantly correlated with TNM-stage and T-stage. CCK-8 assay and colony formation assay demonstrated that LINC02310 acted as an enhancer in LADC. In addition, 3 targeted miRNAs of LINC02310 and 414 downstream DEmRNAs were predicted. The downstream DEmRNAs were then enriched in 405 Gene Ontology terms and 11 Kyoto Encyclopedia of Genes and Genomes pathways, which revealed their potential functions and mechanisms. The PPI network showed the interactions among the downstream DEmRNAs.
This study verified LINC02310 as an enhancer in LADC and performed comprehensive analyses on its downstream regulatory network, which might benefit LADC prognoses and therapies.</description><identifier>ISSN: 1755-8794</identifier><identifier>EISSN: 1755-8794</identifier><identifier>DOI: 10.1186/s12920-020-00834-6</identifier><identifier>PMID: 33308216</identifier><language>eng</language><publisher>England: BioMed Central Ltd</publisher><subject>Adenocarcinoma ; Adenocarcinoma of Lung - genetics ; Adenocarcinoma of Lung - mortality ; Biomarkers ; Biomarkers, Tumor ; Cancer ; Carcinoma, Non-Small-Cell Lung - genetics ; Carcinoma, Non-Small-Cell Lung - mortality ; ceRNA regulatory network ; Chemotherapy ; Cholecystokinin ; Colonies ; Datasets ; Datasets as Topic ; Development and progression ; Experiments ; Gene expression ; Gene Expression Regulation, Neoplastic ; Gene Ontology ; Gene Regulatory Networks ; Genetic aspects ; Genetic markers ; Genomes ; Health aspects ; Humans ; Identification and classification ; Kaplan-Meier Estimate ; LADC ; LncRNA ; Lung cancer ; Lung cancer, Non-small cell ; Lung Neoplasms - genetics ; Lung Neoplasms - mortality ; Neoplasm Staging ; Non-coding RNA ; Non-small cell lung carcinoma ; Oncology, Experimental ; PPI ; Prognosis ; Protein Interaction Maps ; Radiation therapy ; Ribonucleic acid ; RNA ; RNA, Long Noncoding - genetics ; RNA, Long Noncoding - metabolism ; RNA, Messenger - genetics ; RNA, Neoplasm - genetics ; Sincalide - analysis ; Small cell lung carcinoma ; Survival analysis ; TCGA ; Tumor Stem Cell Assay ; Tumors ; Values</subject><ispartof>BMC medical genomics, 2020-12, Vol.13 (1), p.185-185, Article 185</ispartof><rights>COPYRIGHT 2020 BioMed Central Ltd.</rights><rights>2020. This work is licensed under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>The Author(s) 2021, corrected publication 2021</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c597t-57d387a2e2b965b4725ff087bb579f6e54653ca81e3b31be259b3f4b0431322b3</citedby><cites>FETCH-LOGICAL-c597t-57d387a2e2b965b4725ff087bb579f6e54653ca81e3b31be259b3f4b0431322b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7731780/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2471127531?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33308216$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhao, Wenyuan</creatorcontrib><creatorcontrib>Wang, Jun</creatorcontrib><creatorcontrib>Luo, Qingxi</creatorcontrib><creatorcontrib>Peng, Wei</creatorcontrib><creatorcontrib>Li, Bin</creatorcontrib><creatorcontrib>Wang, Lei</creatorcontrib><creatorcontrib>Zhang, Chunfang</creatorcontrib><creatorcontrib>Duan, Chaojun</creatorcontrib><title>Identification of LINC02310 as an enhancer in lung adenocarcinoma and investigation of its regulatory network via comprehensive analyses</title><title>BMC medical genomics</title><addtitle>BMC Med Genomics</addtitle><description>Lung adenocarcinoma (LADC) is a major subtype of non-small cell lung cancer and has one of the highest mortality rates. An increasing number of long non-coding RNAs (LncRNAs) were reported to be associated with the occurrence and progression of LADC. Thus, it is necessary and reasonable to find new prognostic biomarkers for LADC among LncRNAs.
Differential expression analysis, survival analysis, PCR experiments and clinical feature analysis were performed to screen out the LncRNA which was significantly related to LADC. Its role in LADC was verified by CCK-8 assay and colony. Furthermore, competing endogenous RNA (ceRNA) regulatory network construction, enrichment analysis and protein-protein interaction (PPI) network construction were performed to investigate the downstream regulatory network of the selected LncRNA.
A total of 2431 differentially expressed LncRNAs (DELncRNAs) and 2227 differentially expressed mRNAs (DEmRNAs) were from The Cancer Genome Atlas database. Survival analysis results indicated that lnc-YARS2-5, lnc-NPR3-2 and LINC02310 were significantly related to overall survival. Their overexpression indicated poor prognostic. PCR experiments and clinical feature analysis suggested that LINC02310 was significantly correlated with TNM-stage and T-stage. CCK-8 assay and colony formation assay demonstrated that LINC02310 acted as an enhancer in LADC. In addition, 3 targeted miRNAs of LINC02310 and 414 downstream DEmRNAs were predicted. The downstream DEmRNAs were then enriched in 405 Gene Ontology terms and 11 Kyoto Encyclopedia of Genes and Genomes pathways, which revealed their potential functions and mechanisms. The PPI network showed the interactions among the downstream DEmRNAs.
This study verified LINC02310 as an enhancer in LADC and performed comprehensive analyses on its downstream regulatory network, which might benefit LADC prognoses and therapies.</description><subject>Adenocarcinoma</subject><subject>Adenocarcinoma of Lung - genetics</subject><subject>Adenocarcinoma of Lung - mortality</subject><subject>Biomarkers</subject><subject>Biomarkers, Tumor</subject><subject>Cancer</subject><subject>Carcinoma, Non-Small-Cell Lung - genetics</subject><subject>Carcinoma, Non-Small-Cell Lung - mortality</subject><subject>ceRNA regulatory network</subject><subject>Chemotherapy</subject><subject>Cholecystokinin</subject><subject>Colonies</subject><subject>Datasets</subject><subject>Datasets as Topic</subject><subject>Development and progression</subject><subject>Experiments</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>Gene Ontology</subject><subject>Gene Regulatory Networks</subject><subject>Genetic aspects</subject><subject>Genetic markers</subject><subject>Genomes</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Identification and classification</subject><subject>Kaplan-Meier Estimate</subject><subject>LADC</subject><subject>LncRNA</subject><subject>Lung cancer</subject><subject>Lung cancer, Non-small cell</subject><subject>Lung Neoplasms - genetics</subject><subject>Lung Neoplasms - mortality</subject><subject>Neoplasm Staging</subject><subject>Non-coding RNA</subject><subject>Non-small cell lung carcinoma</subject><subject>Oncology, Experimental</subject><subject>PPI</subject><subject>Prognosis</subject><subject>Protein Interaction Maps</subject><subject>Radiation therapy</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA, Long Noncoding - genetics</subject><subject>RNA, Long Noncoding - metabolism</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Neoplasm - genetics</subject><subject>Sincalide - analysis</subject><subject>Small cell lung carcinoma</subject><subject>Survival analysis</subject><subject>TCGA</subject><subject>Tumor Stem Cell Assay</subject><subject>Tumors</subject><subject>Values</subject><issn>1755-8794</issn><issn>1755-8794</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNptkluP1CAYhhujcdfVP-CFIfFmvejKoUB7Y7KZeJhkoomHa0Lp1w5jCyO0o_MP_NnSmXWyYwwhEHjeF3j5suw5wTeElOJ1JLSiOMdzxyUrcvEguySS87yUVfHw3vwiexLjBmOBeUUeZxeMMVxSIi6z38sG3Ghba_RovUO-RavlxwWmjGCkI9IOgVtrZyAg61A_uQ7pJPFGB2OdH3RCmrS1gzja7mRix4gCdFOvRx_2yMH404fvaGc1Mn7YBliDi3YHSa37fYT4NHvU6j7Cs7vxKvv27u3XxYd89en9cnG7yg2v5Jhz2bBSagq0rgSvC0l52-JS1jWXVSuAF4Izo0sCrGakBsqrmrVFjQtGGKU1u8qWR9_G643aBjvosFdeW3VY8KFTOozW9KCEBEokxSnrqijqqjYF4bJgUoIRjebJ683RazvVAzQmJRl0f2Z6vuPsWnV-p6RkRJY4GVzfGQT_Y0oJqsFGA32vHfgpKlpIjJkUB_TlP-jGTyGFd6AIoZKnF56oTqcHWNf6dK6ZTdWt4On_K1bN9775D5VaA4M13kFr0_qZ4NWZIDEj_Bo7PcWoll8-n7P0yJrgYwzQnvIgWM11q451q_Dc57pVIole3E_yJPlbqOwPyx_miQ</recordid><startdate>20201211</startdate><enddate>20201211</enddate><creator>Zhao, Wenyuan</creator><creator>Wang, Jun</creator><creator>Luo, Qingxi</creator><creator>Peng, Wei</creator><creator>Li, Bin</creator><creator>Wang, Lei</creator><creator>Zhang, Chunfang</creator><creator>Duan, Chaojun</creator><general>BioMed Central Ltd</general><general>BioMed Central</general><general>BMC</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>ISR</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20201211</creationdate><title>Identification of LINC02310 as an enhancer in lung adenocarcinoma and investigation of its regulatory network via comprehensive analyses</title><author>Zhao, Wenyuan ; Wang, Jun ; Luo, Qingxi ; Peng, Wei ; Li, Bin ; Wang, Lei ; Zhang, Chunfang ; Duan, Chaojun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c597t-57d387a2e2b965b4725ff087bb579f6e54653ca81e3b31be259b3f4b0431322b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Adenocarcinoma</topic><topic>Adenocarcinoma of Lung - genetics</topic><topic>Adenocarcinoma of Lung - mortality</topic><topic>Biomarkers</topic><topic>Biomarkers, Tumor</topic><topic>Cancer</topic><topic>Carcinoma, Non-Small-Cell Lung - genetics</topic><topic>Carcinoma, Non-Small-Cell Lung - mortality</topic><topic>ceRNA regulatory network</topic><topic>Chemotherapy</topic><topic>Cholecystokinin</topic><topic>Colonies</topic><topic>Datasets</topic><topic>Datasets as Topic</topic><topic>Development and progression</topic><topic>Experiments</topic><topic>Gene expression</topic><topic>Gene Expression Regulation, Neoplastic</topic><topic>Gene Ontology</topic><topic>Gene Regulatory Networks</topic><topic>Genetic aspects</topic><topic>Genetic markers</topic><topic>Genomes</topic><topic>Health aspects</topic><topic>Humans</topic><topic>Identification and classification</topic><topic>Kaplan-Meier Estimate</topic><topic>LADC</topic><topic>LncRNA</topic><topic>Lung cancer</topic><topic>Lung cancer, Non-small cell</topic><topic>Lung Neoplasms - genetics</topic><topic>Lung Neoplasms - mortality</topic><topic>Neoplasm Staging</topic><topic>Non-coding RNA</topic><topic>Non-small cell lung carcinoma</topic><topic>Oncology, Experimental</topic><topic>PPI</topic><topic>Prognosis</topic><topic>Protein Interaction Maps</topic><topic>Radiation therapy</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>RNA, Long Noncoding - genetics</topic><topic>RNA, Long Noncoding - metabolism</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Neoplasm - genetics</topic><topic>Sincalide - analysis</topic><topic>Small cell lung carcinoma</topic><topic>Survival analysis</topic><topic>TCGA</topic><topic>Tumor Stem Cell Assay</topic><topic>Tumors</topic><topic>Values</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhao, Wenyuan</creatorcontrib><creatorcontrib>Wang, Jun</creatorcontrib><creatorcontrib>Luo, Qingxi</creatorcontrib><creatorcontrib>Peng, Wei</creatorcontrib><creatorcontrib>Li, Bin</creatorcontrib><creatorcontrib>Wang, Lei</creatorcontrib><creatorcontrib>Zhang, Chunfang</creatorcontrib><creatorcontrib>Duan, Chaojun</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 In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>ProQuest Health and Medical</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</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)</collection><collection>ProQuest Central</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>ProQuest Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Publicly Available Content Database (Proquest) (PQ_SDU_P3)</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>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>BMC medical genomics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhao, Wenyuan</au><au>Wang, Jun</au><au>Luo, Qingxi</au><au>Peng, Wei</au><au>Li, Bin</au><au>Wang, Lei</au><au>Zhang, Chunfang</au><au>Duan, Chaojun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of LINC02310 as an enhancer in lung adenocarcinoma and investigation of its regulatory network via comprehensive analyses</atitle><jtitle>BMC medical genomics</jtitle><addtitle>BMC Med Genomics</addtitle><date>2020-12-11</date><risdate>2020</risdate><volume>13</volume><issue>1</issue><spage>185</spage><epage>185</epage><pages>185-185</pages><artnum>185</artnum><issn>1755-8794</issn><eissn>1755-8794</eissn><abstract>Lung adenocarcinoma (LADC) is a major subtype of non-small cell lung cancer and has one of the highest mortality rates. An increasing number of long non-coding RNAs (LncRNAs) were reported to be associated with the occurrence and progression of LADC. Thus, it is necessary and reasonable to find new prognostic biomarkers for LADC among LncRNAs.
Differential expression analysis, survival analysis, PCR experiments and clinical feature analysis were performed to screen out the LncRNA which was significantly related to LADC. Its role in LADC was verified by CCK-8 assay and colony. Furthermore, competing endogenous RNA (ceRNA) regulatory network construction, enrichment analysis and protein-protein interaction (PPI) network construction were performed to investigate the downstream regulatory network of the selected LncRNA.
A total of 2431 differentially expressed LncRNAs (DELncRNAs) and 2227 differentially expressed mRNAs (DEmRNAs) were from The Cancer Genome Atlas database. Survival analysis results indicated that lnc-YARS2-5, lnc-NPR3-2 and LINC02310 were significantly related to overall survival. Their overexpression indicated poor prognostic. PCR experiments and clinical feature analysis suggested that LINC02310 was significantly correlated with TNM-stage and T-stage. CCK-8 assay and colony formation assay demonstrated that LINC02310 acted as an enhancer in LADC. In addition, 3 targeted miRNAs of LINC02310 and 414 downstream DEmRNAs were predicted. The downstream DEmRNAs were then enriched in 405 Gene Ontology terms and 11 Kyoto Encyclopedia of Genes and Genomes pathways, which revealed their potential functions and mechanisms. The PPI network showed the interactions among the downstream DEmRNAs.
This study verified LINC02310 as an enhancer in LADC and performed comprehensive analyses on its downstream regulatory network, which might benefit LADC prognoses and therapies.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>33308216</pmid><doi>10.1186/s12920-020-00834-6</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Adenocarcinoma Adenocarcinoma of Lung - genetics Adenocarcinoma of Lung - mortality Biomarkers Biomarkers, Tumor Cancer Carcinoma, Non-Small-Cell Lung - genetics Carcinoma, Non-Small-Cell Lung - mortality ceRNA regulatory network Chemotherapy Cholecystokinin Colonies Datasets Datasets as Topic Development and progression Experiments Gene expression Gene Expression Regulation, Neoplastic Gene Ontology Gene Regulatory Networks Genetic aspects Genetic markers Genomes Health aspects Humans Identification and classification Kaplan-Meier Estimate LADC LncRNA Lung cancer Lung cancer, Non-small cell Lung Neoplasms - genetics Lung Neoplasms - mortality Neoplasm Staging Non-coding RNA Non-small cell lung carcinoma Oncology, Experimental PPI Prognosis Protein Interaction Maps Radiation therapy Ribonucleic acid RNA RNA, Long Noncoding - genetics RNA, Long Noncoding - metabolism RNA, Messenger - genetics RNA, Neoplasm - genetics Sincalide - analysis Small cell lung carcinoma Survival analysis TCGA Tumor Stem Cell Assay Tumors Values |
title | Identification of LINC02310 as an enhancer in lung adenocarcinoma and investigation of its regulatory network via comprehensive analyses |
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