Integrated analysis of gene expression and methylation profiles of novel pancreatic cancer cell lines with highly metastatic activity
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摘要
Pancreatic cancer is one of the most lethal human malignancies, partly because of its propensity for metastasis. However, highly metastatic human pancreatic cancer cell lines suitable for studies of metastasis are currently lacking. Here we established two highly metastatic human pancreatic cancer cell lines, MIA PaCa-2 In8 and Panc-1 In8, by Matrigel induction assay. The cell lines were further characterized both in vitro and in vivo. MIA PaCa-2 In8 and Panc-1 In8 cells demonstrated increased migration and invasion compared with their respective parental cells. Following injection into nude mice, MIA PaCa-2 In8 and Panc-1 In8 cells resulted in more pulmonary metastases compared with the parental cells. Furthermore, analyses of m RNA, long non-coding RNA, micro RNA, and methylation profiling revealed that these factors were aberrantly regulated in the highly metastatic cells,indicating that they probably affected metastasis. We thus established and characterized two highly metastatic human pancreatic cell lines that could be used as valuable tools for future investigations into the pathogenesis, metastasis, and potential treatment of human pancreatic cancer.
Pancreatic cancer is one of the most lethal human malignancies, partly because of its propensity for metastasis. However, highly metastatic human pancreatic cancer cell lines suitable for studies of metastasis are currently lacking. Here we established two highly metastatic human pancreatic cancer cell lines, MIA PaCa-2 In8 and Panc-1 In8, by Matrigel induction assay. The cell lines were further characterized both in vitro and in vivo. MIA PaCa-2 In8 and Panc-1 In8 cells demonstrated increased migration and invasion compared with their respective parental cells. Following injection into nude mice, MIA PaCa-2 In8 and Panc-1 In8 cells resulted in more pulmonary metastases compared with the parental cells. Furthermore, analyses of m RNA, long non-coding RNA, micro RNA, and methylation profiling revealed that these factors were aberrantly regulated in the highly metastatic cells,indicating that they probably affected metastasis. We thus established and characterized two highly metastatic human pancreatic cell lines that could be used as valuable tools for future investigations into the pathogenesis, metastasis, and potential treatment of human pancreatic cancer.
Pancreatic cancer is one of the most lethal human malignancies, partly because of its propensity for metastasis. However, highly metastatic human pancreatic cancer cell lines suitable for studies of metastasis are currently lacking. Here we established two highly metastatic human pancreatic cancer cell lines, MIA PaCa-2 In8 and Panc-1 In8, by Matrigel induction assay. The cell lines were further characterized both in vitro and in vivo. MIA PaCa-2 In8 and Panc-1 In8 cells demonstrated increased migration and invasion compared with their respective parental cells. Following injection into nude mice, MIA PaCa-2 In8 and Panc-1 In8 cells resulted in more pulmonary metastases compared with the parental cells. Furthermore, analyses of m RNA, long non-coding RNA, micro RNA, and methylation profiling revealed that these factors were aberrantly regulated in the highly metastatic cells,indicating that they probably affected metastasis. We thus established and characterized two highly metastatic human pancreatic cell lines that could be used as valuable tools for future investigations into the pathogenesis, metastasis, and potential treatment of human pancreatic cancer.
引文
Ahmad,I.,Mui,E.,Galbraith,L.,Patel,R.,Tan,E.H.,Salji,M.,Rust,A.G.,Repiscak,P.,Hedley,A.,Markert,E.,et al.(2016).Sleeping Beauty screen reveals Pparg activation in metastatic prostate cancer.Proc Natl Acad Sci USA 113,8290-8295.
Bailey,P.,Chang,D.K.,Nones,K.,Johns,A.L.,Patch,A.M.,Gingras,M.C.,Miller,D.K.,Christ,A.N.,Bruxner,T.J.C.,Quinn,M.C.,et al.(2016).Genomic analyses identify molecular subtypes of pancreatic cancer.Nature 531,47-52.
Chao,Y.C.,Pan,S.H.,Yang,S.C.,Yu,S.L.,Che,T.F.,Lin,C.W.,Tsai,M.S.,Chang,G.C.,Wu,C.H.,Wu,Y.Y.,et al.(2009).Claudin-1 is a metastasis suppressor and correlates with clinical outcome in lung adenocarcinoma.Am J Respir Crit Care Med 179,123-133.
Du,Y.,Liu,Z.,You,L.,Hou,P.,Ren,X.,Jiao,T.,Zhao,W.,Li,Z.,Shu,H.,Liu,C.,et al.(2017).Pancreatic cancer progression relies upon mutant p53-induced oncogenic signaling mediated by NOP14.Cancer Res 77,2661-2673.
Gilkes,D.M.,Semenza,G.L.,and Wirtz,D.(2014).Hypoxia and the extracellular matrix:drivers of tumour metastasis.Nat Rev Cancer 14,430-439.
Giovannetti,E.,van der Borden,C.L.,Frampton,A.E.,Ali,A.,Firuzi,O.,and Peters,G.J.(2017).Never let it go:stopping key mechanisms underlying metastasis to fight pancreatic cancer.Seminars Cancer Biol44,43-59.
Go,K.L.,Delitto,D.,Judge,S.M.,Gerber,M.H.,George Jr,T.J.,Behrns,K.E.,Hughes,S.J.,Judge,A.R.,and Trevino,J.G.(2017).Orthotopic patient-derived pancreatic cancer xenografts engraft into the pancreatic parenchyma,metastasize,and induce muscle wasting to recapitulate the human disease.Pancreas 46,813-819.
Goldstein,J.T.,Berger,A.C.,Shih,J.,Duke,F.F.,Furst,L.,Kwiatkowski,D.J.,Cherniack,A.D.,Meyerson,M.,and Strathdee,C.A.(2017).Genomic activation of PPARG reveals a candidate therapeutic axis in bladder cancer.Cancer Res 77,6987-6998.
Grenman,R.,Burk,D.,Virolainen,E.,Buick,R.N.,Church,J.,Schwartz,D.R.,and Carey,T.E.(1989).Clonogenic cell assay for anchoragedependent squamous carcinoma cell lines using limiting dilution.Int JCancer 44,131-136.
Haskins,J.W.,Nguyen,D.X.,and Stern,D.F.(2014).Neuregulin 1-activated ERBB4 interacts with YAP to induce Hippo pathway target genes and promote cell migration.Sci Signal 7,ra116.
Hatano,H.,Kudo,Y.,Ogawa,I.,Tsunematsu,T.,Kikuchi,A.,Abiko,Y.,and Takata,T.(2008).IFN-induced transmembrane protein 1 promotes invasion at early stage of head and neck cancer progression.Clin Cancer Res 14,6097-6105.
Hsu,S.D.,Tseng,Y.T.,Shrestha,S.,Lin,Y.L.,Khaleel,A.,Chou,C.H.,Chu,C.F.,Huang,H.Y.,Lin,C.M.,Ho,S.Y.,et al.(2014).miRTarBase update 2014:an information resource for experimentally validated miRNA-target interactions.Nucl Acids Res 42,D78-D85.
Huang,J.S.,Egger,M.E.,Grizzle,W.E.,and McNally,L.R.(2013).MicroRNA-100 regulates IGF1-receptor expression in metastatic pancreatic cancer cells.Biotech Histochem 88,397-402.
Jeggari,A.,Marks,D.S.,and Larsson,E.(2012).miRcode:a map of putative microRNA target sites in the long non-coding transcriptome.Bioinformatics 28,2062-2063.
Karreth,F.A.,and Pandolfi,P.P.(2013).ceRNA cross-talk in cancer:when ce-bling rivalries go awry.Cancer Discov 3,1113-1121.
Kim,N.H.,Sung,H.Y.,Choi,E.N.,Lyu,D.,Choi,H.J.,Ju,W.,and Ahn,J.H.(2014).Aberrant DNA methylation in the IFITM1 promoter enhances the metastatic phenotype in an intraperitoneal xenograft model of human ovarian cancer.Oncol Rep 31,2139-2146.
Kiuchi,T.,Ortiz-Zapater,E.,Monypenny,J.,Matthews,D.R.,Nguyen,L.K.,Barbeau,J.,Coban,O.,Lawler,K.,Burford,B.,Rolfe,D.J.,et al.(2014).The ErbB4 CYT2 variant protects EGFR from ligand-induced degradation to enhance cancer cell motility.Sci Signal 7,ra78.
Li,D.,Peng,Z.,Tang,H.,Wei,P.,Kong,X.,Yan,D.,Huang,F.,Li,Q.,Le,X.,Li,Q.,et al.(2011).KLF4-mediated negative regulation of IFITM3expression plays a critical role in colon cancer pathogenesis.Clin Cancer Res 17,3558-3568.
Makohon-Moore,A.,and Iacobuzio-Donahue,C.A.(2016).Pancreatic cancer biology and genetics from an evolutionary perspective.Nat Rev Cancer 16,553-565.
McDonald,O.G.,Li,X.,Saunders,T.,Tryggvadottir,R.,Mentch,S.J.,Warmoes,M.O.,Word,A.E.,Carrer,A.,Salz,T.H.,Natsume,S.,et al.(2017).Epigenomic reprogramming during pancreatic cancer progression links anabolic glucose metabolism to distant metastasis.Nat Genet 49,367-376.
Nishimori,H.,Yasoshima,T.,Denno,R.,Shishido,T.,Hata,F.,Honma,T.,Ura,H.,Yamaguchi,K.,Yagihashi,A.,Tanaka,H.,et al.(2001).A new peritoneal dissemination model established from the human pancreatic cancer cell line.Pancreas 22,348-356.
Nishimori,H.,Yasoshima,T.,Hata,F.,Denno,R.,Yanai,Y.,Nomura,H.,Tanaka,H.,Kamiguchi,K.,Sato,N.,and Hirata,K.(2002).A novel nude mouse model of liver metastasis and peritoneal dissemination from the same human pancreatic cancer line.Pancreas 24,242-250.
Notta,F.,Chan-Seng-Yue,M.,Lemire,M.,Li,Y.,Wilson,G.W.,Connor,A.A.,Denroche,R.E.,Liang,S.B.,Brown,A.M.K.,Kim,J.C.,et al.(2016).A renewed model of pancreatic cancer evolution based on genomic rearrangement patterns.Nature 538,378-382.
Ottaviani,S.,Stebbing,J.,Frampton,A.E.,Zagorac,S.,Krell,J.,de Giorgio,A.,Trabulo,S.M.,Nguyen,V.T.M.,Magnani,L.,Feng,H.,et al.(2018).TGF-βinduces miR-100 and miR-125b but blocks let-7a through LIN28B controlling PDAC progression.Nat Commun 9,1845.
Pertea,M.,Kim,D.,Pertea,G.M.,Leek,J.T.,and Salzberg,S.L.(2016).Transcript-level expression analysis of RNA-seq experiments with HISAT,StringTie and Ballgown.Nat Protoc 11,1650-1667.
Pertea,M.,Pertea,G.M.,Antonescu,C.M.,Chang,T.C.,Mendell,J.T.,and Salzberg,S.L.(2015).StringTie enables improved reconstruction of a transcriptome from RNA-seq reads.Nat Biotechnol 33,290-295.
Rahib,L.,Smith,B.D.,Aizenberg,R.,Rosenzweig,A.B.,Fleshman,J.M.,and Matrisian,L.M.(2014).Projecting cancer incidence and deaths to2030:the unexpected burden of thyroid,liver,and pancreas cancers in the United States.Cancer Res 74,2913-2921.
Reticker-Flynn,N.E.,Malta,D.F.B.,Winslow,M.M.,Lamar,J.M.,Xu,M.J.,Underhill,G.H.,Hynes,R.O.,Jacks,T.E.,and Bhatia,S.N.(2012).Acombinatorial extracellular matrix platform identifies cell-extracellular matrix interactions that correlate with metastasis.Nat Commun 3,1122.
Salameh,A.,Lee,A.K.,Cardó-Vila,M.,Nunes,D.N.,Efstathiou,E.,Staquicini,F.I.,Dobroff,A.S.,Marchiò,S.,Navone,N.M.,Hosoya,H.,et al.(2015).PRUNE2 is a human prostate cancer suppressor regulated by the intronic long noncoding RNA PCA3.Proc Natl Acad Sci USA112,8403-8408.
Settleman,J.,Lee,A.K.,Cardo-Vila,M.,Nunes,D.N.,Efstathiou,E.,Staquicini,F.I.,Dobroff,A.S.,Marchio,S.,Navone,N.M.,Hosoya,H.,et al.(2009).A therapeutic opportunity in melanoma:ErbB4 makes a mark on skin.Cancer Cell 16,278-279.
Shannon,P.,Markiel,A.,Ozier,O.,Baliga,N.S.,Wang,J.T.,Ramage,D.,Amin,N.,Schwikowski,B.,and Ideker,T.(2003).Cytoscape:a software environment for integrated models of biomolecular interaction networks.Genome Res 13,2498-2504.
Siegel,R.L.,Miller,K.D.,and Jemal,A.(2018).Cancer statistics,2018.CA-A Cancer J Clin 68,7-30.
Srivastava,S.K.,Arora,S.,Singh,S.,Bhardwaj,A.,Averett,C.,and Singh,A.P.(2014).MicroRNAs in pancreatic malignancy:progress and promises.Cancer Lett 347,167-174.
Tahira,A.C.,Kubrusly,M.S.,Faria,M.F.,Dazzani,B.,Fonseca,R.S.,Maracaja-Coutinho,V.,Verjovski-Almeida,S.,Machado,M.C.C.,and Reis,E.M.(2011).Long noncoding intronic RNAs are differentially expressed in primary and metastatic pancreatic cancer.Mol Cancer 10,141.
Wang,C.,Zhang,W.,Zhang,L.,Chen,X.,Liu,F.,Zhang,J.,Guan,S.,Sun,Y.,Chen,P.,Wang,D.,et al.(2016).miR-146a-5p mediates epithelialmesenchymal transition of oesophageal squamous cell carcinoma via targeting Notch2.Br J Cancer 115,1548-1554.
Wang,P.,Chen,L.,Zhang,J.,Chen,H.,Fan,J.,Wang,K.,Luo,J.,Chen,Z.,Meng,Z.,and Liu,L.(2014).Methylation-mediated silencing of the miR-124 genes facilitates pancreatic cancer progression and metastasis by targeting Rac1.Oncogene 33,514-524.
Yang,Q.,Wang,Y.,Lu,X.,Zhao,Z.,Zhu,L.,Chen,S.,Wu,Q.,Chen,C.,and Wang,Z.(2015).MiR-125b regulates epithelial-mesenchymal transition via targeting Sema4C in paclitaxel-resistant breast cancer cells.Oncotarget 6,3268-3279.
Zhang,J.Y.,Weng,M.Z.,Song,F.B.,Xu,Y.G.,Liu,Q.,Wu,J.Y.,Qin,J.,Jin,T.,and Xu,J.M.(2016).Long noncoding RNA AFAP1-AS1indicates a poor prognosis of hepatocellular carcinoma and promotes cell proliferation and invasion via upregulation of the RhoA/Rac2signaling.Int J Oncol 48,1590-1598.
Bailey,P.,Chang,D.K.,Nones,K.,Johns,A.L.,Patch,A.M.,Gingras,M.C.,Miller,D.K.,Christ,A.N.,Bruxner,T.J.C.,Quinn,M.C.,et al.(2016).Genomic analyses identify molecular subtypes of pancreatic cancer.Nature 531,47-52.
Chao,Y.C.,Pan,S.H.,Yang,S.C.,Yu,S.L.,Che,T.F.,Lin,C.W.,Tsai,M.S.,Chang,G.C.,Wu,C.H.,Wu,Y.Y.,et al.(2009).Claudin-1 is a metastasis suppressor and correlates with clinical outcome in lung adenocarcinoma.Am J Respir Crit Care Med 179,123-133.
Du,Y.,Liu,Z.,You,L.,Hou,P.,Ren,X.,Jiao,T.,Zhao,W.,Li,Z.,Shu,H.,Liu,C.,et al.(2017).Pancreatic cancer progression relies upon mutant p53-induced oncogenic signaling mediated by NOP14.Cancer Res 77,2661-2673.
Gilkes,D.M.,Semenza,G.L.,and Wirtz,D.(2014).Hypoxia and the extracellular matrix:drivers of tumour metastasis.Nat Rev Cancer 14,430-439.
Giovannetti,E.,van der Borden,C.L.,Frampton,A.E.,Ali,A.,Firuzi,O.,and Peters,G.J.(2017).Never let it go:stopping key mechanisms underlying metastasis to fight pancreatic cancer.Seminars Cancer Biol44,43-59.
Go,K.L.,Delitto,D.,Judge,S.M.,Gerber,M.H.,George Jr,T.J.,Behrns,K.E.,Hughes,S.J.,Judge,A.R.,and Trevino,J.G.(2017).Orthotopic patient-derived pancreatic cancer xenografts engraft into the pancreatic parenchyma,metastasize,and induce muscle wasting to recapitulate the human disease.Pancreas 46,813-819.
Goldstein,J.T.,Berger,A.C.,Shih,J.,Duke,F.F.,Furst,L.,Kwiatkowski,D.J.,Cherniack,A.D.,Meyerson,M.,and Strathdee,C.A.(2017).Genomic activation of PPARG reveals a candidate therapeutic axis in bladder cancer.Cancer Res 77,6987-6998.
Grenman,R.,Burk,D.,Virolainen,E.,Buick,R.N.,Church,J.,Schwartz,D.R.,and Carey,T.E.(1989).Clonogenic cell assay for anchoragedependent squamous carcinoma cell lines using limiting dilution.Int JCancer 44,131-136.
Haskins,J.W.,Nguyen,D.X.,and Stern,D.F.(2014).Neuregulin 1-activated ERBB4 interacts with YAP to induce Hippo pathway target genes and promote cell migration.Sci Signal 7,ra116.
Hatano,H.,Kudo,Y.,Ogawa,I.,Tsunematsu,T.,Kikuchi,A.,Abiko,Y.,and Takata,T.(2008).IFN-induced transmembrane protein 1 promotes invasion at early stage of head and neck cancer progression.Clin Cancer Res 14,6097-6105.
Hsu,S.D.,Tseng,Y.T.,Shrestha,S.,Lin,Y.L.,Khaleel,A.,Chou,C.H.,Chu,C.F.,Huang,H.Y.,Lin,C.M.,Ho,S.Y.,et al.(2014).miRTarBase update 2014:an information resource for experimentally validated miRNA-target interactions.Nucl Acids Res 42,D78-D85.
Huang,J.S.,Egger,M.E.,Grizzle,W.E.,and McNally,L.R.(2013).MicroRNA-100 regulates IGF1-receptor expression in metastatic pancreatic cancer cells.Biotech Histochem 88,397-402.
Jeggari,A.,Marks,D.S.,and Larsson,E.(2012).miRcode:a map of putative microRNA target sites in the long non-coding transcriptome.Bioinformatics 28,2062-2063.
Karreth,F.A.,and Pandolfi,P.P.(2013).ceRNA cross-talk in cancer:when ce-bling rivalries go awry.Cancer Discov 3,1113-1121.
Kim,N.H.,Sung,H.Y.,Choi,E.N.,Lyu,D.,Choi,H.J.,Ju,W.,and Ahn,J.H.(2014).Aberrant DNA methylation in the IFITM1 promoter enhances the metastatic phenotype in an intraperitoneal xenograft model of human ovarian cancer.Oncol Rep 31,2139-2146.
Kiuchi,T.,Ortiz-Zapater,E.,Monypenny,J.,Matthews,D.R.,Nguyen,L.K.,Barbeau,J.,Coban,O.,Lawler,K.,Burford,B.,Rolfe,D.J.,et al.(2014).The ErbB4 CYT2 variant protects EGFR from ligand-induced degradation to enhance cancer cell motility.Sci Signal 7,ra78.
Li,D.,Peng,Z.,Tang,H.,Wei,P.,Kong,X.,Yan,D.,Huang,F.,Li,Q.,Le,X.,Li,Q.,et al.(2011).KLF4-mediated negative regulation of IFITM3expression plays a critical role in colon cancer pathogenesis.Clin Cancer Res 17,3558-3568.
Makohon-Moore,A.,and Iacobuzio-Donahue,C.A.(2016).Pancreatic cancer biology and genetics from an evolutionary perspective.Nat Rev Cancer 16,553-565.
McDonald,O.G.,Li,X.,Saunders,T.,Tryggvadottir,R.,Mentch,S.J.,Warmoes,M.O.,Word,A.E.,Carrer,A.,Salz,T.H.,Natsume,S.,et al.(2017).Epigenomic reprogramming during pancreatic cancer progression links anabolic glucose metabolism to distant metastasis.Nat Genet 49,367-376.
Nishimori,H.,Yasoshima,T.,Denno,R.,Shishido,T.,Hata,F.,Honma,T.,Ura,H.,Yamaguchi,K.,Yagihashi,A.,Tanaka,H.,et al.(2001).A new peritoneal dissemination model established from the human pancreatic cancer cell line.Pancreas 22,348-356.
Nishimori,H.,Yasoshima,T.,Hata,F.,Denno,R.,Yanai,Y.,Nomura,H.,Tanaka,H.,Kamiguchi,K.,Sato,N.,and Hirata,K.(2002).A novel nude mouse model of liver metastasis and peritoneal dissemination from the same human pancreatic cancer line.Pancreas 24,242-250.
Notta,F.,Chan-Seng-Yue,M.,Lemire,M.,Li,Y.,Wilson,G.W.,Connor,A.A.,Denroche,R.E.,Liang,S.B.,Brown,A.M.K.,Kim,J.C.,et al.(2016).A renewed model of pancreatic cancer evolution based on genomic rearrangement patterns.Nature 538,378-382.
Ottaviani,S.,Stebbing,J.,Frampton,A.E.,Zagorac,S.,Krell,J.,de Giorgio,A.,Trabulo,S.M.,Nguyen,V.T.M.,Magnani,L.,Feng,H.,et al.(2018).TGF-βinduces miR-100 and miR-125b but blocks let-7a through LIN28B controlling PDAC progression.Nat Commun 9,1845.
Pertea,M.,Kim,D.,Pertea,G.M.,Leek,J.T.,and Salzberg,S.L.(2016).Transcript-level expression analysis of RNA-seq experiments with HISAT,StringTie and Ballgown.Nat Protoc 11,1650-1667.
Pertea,M.,Pertea,G.M.,Antonescu,C.M.,Chang,T.C.,Mendell,J.T.,and Salzberg,S.L.(2015).StringTie enables improved reconstruction of a transcriptome from RNA-seq reads.Nat Biotechnol 33,290-295.
Rahib,L.,Smith,B.D.,Aizenberg,R.,Rosenzweig,A.B.,Fleshman,J.M.,and Matrisian,L.M.(2014).Projecting cancer incidence and deaths to2030:the unexpected burden of thyroid,liver,and pancreas cancers in the United States.Cancer Res 74,2913-2921.
Reticker-Flynn,N.E.,Malta,D.F.B.,Winslow,M.M.,Lamar,J.M.,Xu,M.J.,Underhill,G.H.,Hynes,R.O.,Jacks,T.E.,and Bhatia,S.N.(2012).Acombinatorial extracellular matrix platform identifies cell-extracellular matrix interactions that correlate with metastasis.Nat Commun 3,1122.
Salameh,A.,Lee,A.K.,Cardó-Vila,M.,Nunes,D.N.,Efstathiou,E.,Staquicini,F.I.,Dobroff,A.S.,Marchiò,S.,Navone,N.M.,Hosoya,H.,et al.(2015).PRUNE2 is a human prostate cancer suppressor regulated by the intronic long noncoding RNA PCA3.Proc Natl Acad Sci USA112,8403-8408.
Settleman,J.,Lee,A.K.,Cardo-Vila,M.,Nunes,D.N.,Efstathiou,E.,Staquicini,F.I.,Dobroff,A.S.,Marchio,S.,Navone,N.M.,Hosoya,H.,et al.(2009).A therapeutic opportunity in melanoma:ErbB4 makes a mark on skin.Cancer Cell 16,278-279.
Shannon,P.,Markiel,A.,Ozier,O.,Baliga,N.S.,Wang,J.T.,Ramage,D.,Amin,N.,Schwikowski,B.,and Ideker,T.(2003).Cytoscape:a software environment for integrated models of biomolecular interaction networks.Genome Res 13,2498-2504.
Siegel,R.L.,Miller,K.D.,and Jemal,A.(2018).Cancer statistics,2018.CA-A Cancer J Clin 68,7-30.
Srivastava,S.K.,Arora,S.,Singh,S.,Bhardwaj,A.,Averett,C.,and Singh,A.P.(2014).MicroRNAs in pancreatic malignancy:progress and promises.Cancer Lett 347,167-174.
Tahira,A.C.,Kubrusly,M.S.,Faria,M.F.,Dazzani,B.,Fonseca,R.S.,Maracaja-Coutinho,V.,Verjovski-Almeida,S.,Machado,M.C.C.,and Reis,E.M.(2011).Long noncoding intronic RNAs are differentially expressed in primary and metastatic pancreatic cancer.Mol Cancer 10,141.
Wang,C.,Zhang,W.,Zhang,L.,Chen,X.,Liu,F.,Zhang,J.,Guan,S.,Sun,Y.,Chen,P.,Wang,D.,et al.(2016).miR-146a-5p mediates epithelialmesenchymal transition of oesophageal squamous cell carcinoma via targeting Notch2.Br J Cancer 115,1548-1554.
Wang,P.,Chen,L.,Zhang,J.,Chen,H.,Fan,J.,Wang,K.,Luo,J.,Chen,Z.,Meng,Z.,and Liu,L.(2014).Methylation-mediated silencing of the miR-124 genes facilitates pancreatic cancer progression and metastasis by targeting Rac1.Oncogene 33,514-524.
Yang,Q.,Wang,Y.,Lu,X.,Zhao,Z.,Zhu,L.,Chen,S.,Wu,Q.,Chen,C.,and Wang,Z.(2015).MiR-125b regulates epithelial-mesenchymal transition via targeting Sema4C in paclitaxel-resistant breast cancer cells.Oncotarget 6,3268-3279.
Zhang,J.Y.,Weng,M.Z.,Song,F.B.,Xu,Y.G.,Liu,Q.,Wu,J.Y.,Qin,J.,Jin,T.,and Xu,J.M.(2016).Long noncoding RNA AFAP1-AS1indicates a poor prognosis of hepatocellular carcinoma and promotes cell proliferation and invasion via upregulation of the RhoA/Rac2signaling.Int J Oncol 48,1590-1598.