Hepatitis B virus X protein enhances hepatocarcinogenesis by depressing the targeting of NUSAP1 mRNA by miR-18b
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摘要
Objective: The aim of this study was to investigate the underlying mechanism whereby HBx modulates the targeting of NUSAP1 by miR-18b to enhance hepatocarcinogenesis.Methods: We employed an integrated approach of bioinformatics analysis and molecular experiments in hepatoma cells, HBV transgenic mice, and clinical liver cancer tissues to investigate the role of HBx-regulated miR-18b in the development of liver cancer.Results: In this study, we report that the HBx-mediated tumor suppressor miR-18b modulates hepatocarcinogenesis during the host-HBV interaction. The expression levels of miR-18b were lower in clinical HBV-positive liver cancer tissues and liver tissues of HBV-transgenic mice. Interestingly, HBx inhibited miR-18b expression by inducing the methylation of CpG islands in its promoter. Accordingly, we tested the hypothesis that HBx enhanced hepatocarcinogenesis by increasing the expression of target genes of miR-18b. Moreover, we identified nucleolar spindle-associated protein 1(NUSAP1) as one of the target genes of miR-18b.NUSAP1 was expressed at high levels in liver cancer tissues. Interestingly, HBx up-regulated NUSAP1 by suppressing miR-18b.Functionally, miR-18b significantly inhibited the proliferation of hepatoma cells by depressing NUSAP1 levels in vivo and in vitro.Conclusions: Thus, we conclude that the targeting of NUSAP1 mRNA by the tumor suppressor miR-18b is controlled by HBxmodulated promoter methylation during the host-virus interaction, leading to hepatocarcinogenesis. Our findings provide new insights into the mechanism by which HBx-mediated miRNAs modulate hepatocarcinogenesis.
Objective: The aim of this study was to investigate the underlying mechanism whereby HBx modulates the targeting of NUSAP1 by miR-18b to enhance hepatocarcinogenesis.Methods: We employed an integrated approach of bioinformatics analysis and molecular experiments in hepatoma cells, HBV transgenic mice, and clinical liver cancer tissues to investigate the role of HBx-regulated miR-18b in the development of liver cancer.Results: In this study, we report that the HBx-mediated tumor suppressor miR-18b modulates hepatocarcinogenesis during the host-HBV interaction. The expression levels of miR-18b were lower in clinical HBV-positive liver cancer tissues and liver tissues of HBV-transgenic mice. Interestingly, HBx inhibited miR-18b expression by inducing the methylation of CpG islands in its promoter. Accordingly, we tested the hypothesis that HBx enhanced hepatocarcinogenesis by increasing the expression of target genes of miR-18b. Moreover, we identified nucleolar spindle-associated protein 1(NUSAP1) as one of the target genes of miR-18b.NUSAP1 was expressed at high levels in liver cancer tissues. Interestingly, HBx up-regulated NUSAP1 by suppressing miR-18b.Functionally, miR-18b significantly inhibited the proliferation of hepatoma cells by depressing NUSAP1 levels in vivo and in vitro.Conclusions: Thus, we conclude that the targeting of NUSAP1 mRNA by the tumor suppressor miR-18b is controlled by HBxmodulated promoter methylation during the host-virus interaction, leading to hepatocarcinogenesis. Our findings provide new insights into the mechanism by which HBx-mediated miRNAs modulate hepatocarcinogenesis.
Objective: The aim of this study was to investigate the underlying mechanism whereby HBx modulates the targeting of NUSAP1 by miR-18b to enhance hepatocarcinogenesis.Methods: We employed an integrated approach of bioinformatics analysis and molecular experiments in hepatoma cells, HBV transgenic mice, and clinical liver cancer tissues to investigate the role of HBx-regulated miR-18b in the development of liver cancer.Results: In this study, we report that the HBx-mediated tumor suppressor miR-18b modulates hepatocarcinogenesis during the host-HBV interaction. The expression levels of miR-18b were lower in clinical HBV-positive liver cancer tissues and liver tissues of HBV-transgenic mice. Interestingly, HBx inhibited miR-18b expression by inducing the methylation of CpG islands in its promoter. Accordingly, we tested the hypothesis that HBx enhanced hepatocarcinogenesis by increasing the expression of target genes of miR-18b. Moreover, we identified nucleolar spindle-associated protein 1(NUSAP1) as one of the target genes of miR-18b.NUSAP1 was expressed at high levels in liver cancer tissues. Interestingly, HBx up-regulated NUSAP1 by suppressing miR-18b.Functionally, miR-18b significantly inhibited the proliferation of hepatoma cells by depressing NUSAP1 levels in vivo and in vitro.Conclusions: Thus, we conclude that the targeting of NUSAP1 mRNA by the tumor suppressor miR-18b is controlled by HBxmodulated promoter methylation during the host-virus interaction, leading to hepatocarcinogenesis. Our findings provide new insights into the mechanism by which HBx-mediated miRNAs modulate hepatocarcinogenesis.
引文
1. Tsai WL,Chung RT.Viral hepatocarcinogenesis.Oncogene.2010;29:2309-24.
2. Zhang XD,Wang Y,Ye LH.Hepatitis B virus X protein accelerates the development of hepatoma.Cancer Biol Med.2014;11:182-90.
3. Xu C,Zhou WC,Wang YM,Qiao L.Hepatitis B virus-induced hepatocellular carcinoma.Cancer Lett.2014;345:216-22.
4. Sinn DH,Lee J,Goo J,Kim K,Gwak GY,Paik YH,et al.Hepatocellular carcinoma risk in chronic hepatitis B virus-infected compensated cirrhosis patients with low viral load.Hepatology.2015;62:694-701.
5. Arzumanyan A,Sambandam V,Clayton MM,Choi SS,Xie GH,Diehl AM,et al. Hedgehog signaling blockade delays hepatocarcinogenesis induced by hepatitis B virus X protein.Cancer Res.2012;72:5912-20.
6. Zhang T,Zhang JP,You XN,Liu Q,Du YM,Gao YE,et al.Hepatitis B virus x protein modulates oncogene yes-associated protein by CREB to promote growth of hepatoma cells.Hepatology.2012;56:2051-59.
7. Geng X,Huang CH,Qin Y,McCombs JE,Yuan Q,Harry BL,et al.Hepatitis B virus X protein targets Bcl-2 proteins to increase intracellular calcium,required for virus replication and cell death induction.Proc Natl Acad Sci USA.2012;109:18471-6.
8. You XN,Liu FB,Zhang T,Li YH,Ye LH,Zhang XD.Hepatitis B virus X protein upregulates oncogene Rab18 to result in the dysregulation of lipogenesis and proliferatio n of hepatoma cells.Carcinogenesis.2013;34:1644-52.
9. Zhang S,Shan CL,Cui WJ,You XN,Du YM,Kong GY,et al.Hepatitis B virus X protein protects hepatoma and hepatic cells from complement-dependent cytotoxicity by up-regulation of CD46.FEBS Lett.2013;587:645-51.
10. Gao Y,Feng JY,Yang G,Zhang SQ,Liu YX,Bu YN,et al.Hepatitis B virus X protein-elevated MSL2 modulates hepatitis B virus covalently closed circular DNA by inducing degradation of APOBEC3B to enhance hepatocarcinogenesis.Hepatology.2017;66:1413-29.
11. Park ES,Park YK,Shin CY,Park SH,Ahn SH,Kim DH,et al.Hepatitis B virus inhibits liver regeneration via epigenetic regulation of urokinase-type plasminogen activator.Hepatology.2013;58:762-76.
12. Song K,Han C,Wu T.Epigenetic regulation of miR-122 expression by PPAR gamma/RXR alpha complex and HBx in hepatocellular carcinoma.Hepatology.2012;56:609a-10a.
13. Bartel DP.MicroRNAs:genomics,biogenesis,mechanism,and function.Cell.2004;116:281-97.
14. Soifer HS,Rossi JJ,Saetrom P.MicroRNAs in disease and potential therapeutic applications.Mol Ther.2007;15:2070-9.
15. Finnegan EF,Pasquinelli AE.MicroRNA biogenesis:regulating the regulators.Crit Rev Biochem Mol Biol.2013;48:51-68.
16. Gurtan AM,Sharp PA.The role of miRNAs in regulating gene expression networks.J Mol Biol.2013;425:3582-600.
17. Gregory RI,Chendrimada TP,Shiekhattar R.MicroRNA biogenesis:Isolation a nd characterization of the microprocessor complex.In:Ying SY.MicroRNA Protocols.Totowa,NJ:Humana Press;2006;33-47.
18. Jinek M,Doudna JA.A three-dimensional view of the molecular machinery of RNA interference.Nature.2009;457:405-12.
19. Chen RJ,Kelly G,Sengupta A,Heydendael W,Nicholas B,Beltrami S,et al.MicroRNAs as biomarkers of resilience or vulnerability to stress.Neuroscience.2015;305:36-48.
20. Haflidadottir BS,Ceder Y.Exosomal microRNAs as potential biomarkers in castration-resistant prostate cancer.Eur Urol.2015;67:42-3.
21. Kriegel AJ,Baker MA,Liu Y,Liu PY,Cowley AW Jr,Liang MY.Endogenous microRNAs in human microvascular endothelial cells regulate mRNAs encoded by hypertension-related genes.Hypertension.2015;66:793-9.
22. Wang YH,Chen J,Lin ZY,Cao J,Huang HL,Jiang Y,et al.Role of deregulated microRNAs in non-small cell lung cancer progression using fresh-frozen and formalin-fixed,paraffin-embedded samples.Oncol Lett.2016;11:801-8.
23. Chakravarthi BVSK,Pathi SS,Goswami MT,Cieslik M,Zheng H,Nallasivam S,et al.The miR-124-prolyl hydroxylase P4HA1-MMP1axis plays a critical role in prostate cancer progression.Oncotarget.2014;5:6654-69.
24. Tan Gana NH,Onuki T,Victoriano AFB,Okamoto T.MicroRNAs in HIV-1 infection:an integration of viral and cellular interaction at the genomic level.Front Microbiol.2012;3:306.
25. Drury RE,O'Connor D,Pollard AJ.The clinical application of microRNAs in infectious disease.Front Immunol.2017;8:1182.
26. Hou J,Wang P,Lin L,Liu XG,Ma F,An HZ,et al.MicroRNA-146a feedback inhibits RIG-1-dependent TypeⅠIFN production in macrophages by targeting TRAF6,IRAK1,and IRAK2.J Immunol.2009;183:2150-8.
27. Rossato M,Curtale G,Tamassia N,Castellucci M,Mori L,Gasperini S,et al.IL-10-induced micro RNA-187 negatively regulates TNF-α,IL-6,and IL-12p40 production in TLR4-stimulated monocytes.Proc Natl Acad Sci USA.2012;109:E3101-10.
28. Curtale G,Mirolo M,Renzi TA,Rossato M,Bazzoni F,Locati M.Negative regulation of Toll-like receptor 4 signaling by IL-10-dependent microRNA-146b.Proc Natl Acad Sci USA.2013;110:11499-504.
29. Lecellier CH,Dunoyer P,Arar K,Lehmann-Che J,Eyquem S,Himber C,et al.A cellular microRNA mediates antiviral defense in human cells.Science.2005;308:557-60.
30. Nathans R,Chu CY,Serquina AK,Lu CC,Cao H,Rana TM.Cellular microRNA and P bodies modulate host-HIV-1interactions.Mol Cell.2009;34:696-709.
31. Bai XT,Nicot C.miR-28-3p is a cellular restriction factor thatinhibits human T cell leukemia virus,type 1(HTLV-1)replication and virus infection.J Biol Chem.2015;290:5381-90.
32. Hu W,Wang XJ,Ding XR,Li Y,Zhang XJ,Xie PW,et al.MicroRNA-141 represses HBV replication by targeting PPARA.PLoS One.2012;7:e34165.
33. Jin J,Tang SH,Xia L,Du R,Xie HH,Song JG,et al.MicroRNA-501promotes HBV replication by targeting HBXIP.Biochem Biophys Res Commun.2013;430:1228-33.
34. Dar AA,Majid S,Rittsteuer C,de Semir D,Bezrookove V,Tong S,et al.The role of miR-18b in MDM2-p53 pathway signaling and melanoma progression.J Natl Cancer Inst.2013;105:433-42.
35. Wu JH,Wang YH,Wang W,Shen W,Sang YZ,Liu L,et al.MiR-18b suppresses high-glucose-induced proliferation in HRECs by targeting IGF-1/IGF1R signaling pathways.Int J Biochem Cell Biol.2016;73:41-52.
36. Lujambio A,Calin GA,Villanueva A,Ropero S,Sanchez-Cespedes M,Blanco D,et al.A microRNA DNA methylation signature for human cancer metastasis.Proc Natl Acad Sci USA.2008;105:13556-61.
37. Lopez-Serra P,Esteller M.DNA methylation-associated silencing of tumor-suppressor microRNAs in cancer.Oncogene.2012;31:1609-22.
38. Zhang T,Zhang JP,Cui M,Liu FB,You XN,Du YM,et al.Hepatitis B virus X protein inhibits tumor suppressor miR-205through inducing hypermethylation of miR-205 promoter to enhance carcinogenesis.Neoplasia.2013;15:1282-91.
39. Vanden Bosch A,Raemaekers T,Denayer S,Torrekens S,Smets N,Moermans K,et al.NuSAP is essential for chromatin-induced spindle formation during early embryogenesis.J Cell Sci.2010;123:3244-55.
40. Okamoto A,Higo M,Shiiba M,Nakashima D,Koyama T,Miyamoto I,et al.Down-regulation of nucleolar and spindleassociated protein 1(NUSAP1)expression suppresses tumor and cell proliferation and enhances anti-tumor effect of paclitaxel in oral squamous cell carcinoma.PLoS One.2015;10:e0142252.
41. Gordon CA,Gulzar ZG,Brooks JD.NUSAP1 expression is upregulated by loss of RBI in prostate cancer cells.Prostate.2015;75:517-26.
42. Chen LD,Zhuo DQ,Chen JK,Yuan HY.Screening feature genes of lung carcinoma with DNA micro array analysis.Int J Clin Exp Med.2015;8:12161-71.
43. Satow R,Shitashige M,Kanai Y,Takeshita F,Ojima H,Jigami T,et al.Combined functional genome survey of therapeutic targets for hepatocellular carcinoma.Clin Cancer Res.2010;16:2518-28.
44. Liu YW,Xia R,Lu K,Xie M,Yang F,Sun M,et al.LincRNAFEZFIAS1 represses p21 expression to promote gastric cancer proliferation th ro ugh LSD1-Mediated H3K4me2 deme thylation.Mol Cancer.2017;16:39.
45. Xu T,Zhu Y,Xiong YJ,Ge YY,Yun JP,Zhuang SM.MicroRNA-195 suppresses tumorigenicity and regulates G1/S transition of human hepatocellular carcinoma cells.Hepatology.2009;50:113-21.
46. Fang L,Zhang M,Chen L,Xiong H,Ge YK,Lu W,et al.Downregulation of nucleolar and spindle-associated protein 1expression suppresses cell migration,proliferation and invasion in renal cell carcinoma.Oncol Rep.2016;36:1506-16.
47. Bimonte S,Leongito M,Barbieri A,Del Vecchio V,Falco M,Giudice A,et al.The therapeutic targets of miRNA in hepatic cancer stem cells.Stem Cells Int.2016;2016:1065230.
48. Braconi C,Henry JC,Kogure T,Schmittgen T,Patel T.The role of MicroRNAs in human liver cancers.Semin Oncol.2011;38:752-63.
49. Coppola N,Potenza N,Pisaturo M,Mosca N,Tonziello G,Signoriello G,et al.Liver microRNA hsa-miR-125a-5p in HBV chronic infection:correlation with HBV replication and disease progression.PLoS One.2013;8:e65336.
50. Jiang RQ,Deng L,Zhao L,Li XC,Zhang F,Xia YX,et al.miR-22promotes HBV-related hepatocellular carcinoma development in males.Clin Cancer Res.2011;17:5593-603.
51. Khokhar A,Noorali S,Sheraz M,Mahalingham K,Pace DG,Khanani MR,et al.Computational analysis to predict functional role of hsa-miR-3065-3p as an antiviral therapeutic agent for treatment of triple infections:HCV,HIV-1,and HBV.Libyan J Med.2012;7:19774.
52. Li BB,Li DL,Chen C,Liu BH,Xia CY,Wu HJ,et al.Potentials of the elevated circulating miR-185 level as a biomarker for early diagnosis of HBV-related liver fibrosis.Sci Rep.2016;6:34157.
53. Katayama Y,Maeda M,Miyaguchi K,Nemoto S,Yasen M,TanakaS,et al.Identification of pathogenesis-related microRNAs in hepatocellular carcinoma by expression profiling.Oncol Lett.20124:817-23.
54. Zhang ZZ,Liu X,Wang DQ,Teng MK,Niu LW,Huang AL,et al.Hepatitis B virus and hepatocellular carcinoma at the miRNA level World J Gastroenterol.2011;17:3353-8.
55. Chen L,Yang L,Qiao F,Hu X,Li S,Yao L,et al.High levels of nucleolar spindle-associated protein and reduced levels of BRCA1expression predict poor prognosis in triple-negative breast cancer.PLoS One.2015;10:e0140572.
56. Wu XQ,Xu BK,Yang C,Wang WT,Zhong DQ,Zhao Z,et al.Nucleolar and spindle associated protein 1 promotes the aggressiveness of astrocytoma by activating the Hedgehog signaling pathway.J Exp Clin Cancer Res.2017;36:127.
57. Gordon CA,Gong X,Ganesh D,Brooks JD.NUSAP1 promotes invasion and metastasis of prostate cancer.Oncotarget.2017;8:29935-50.
2. Zhang XD,Wang Y,Ye LH.Hepatitis B virus X protein accelerates the development of hepatoma.Cancer Biol Med.2014;11:182-90.
3. Xu C,Zhou WC,Wang YM,Qiao L.Hepatitis B virus-induced hepatocellular carcinoma.Cancer Lett.2014;345:216-22.
4. Sinn DH,Lee J,Goo J,Kim K,Gwak GY,Paik YH,et al.Hepatocellular carcinoma risk in chronic hepatitis B virus-infected compensated cirrhosis patients with low viral load.Hepatology.2015;62:694-701.
5. Arzumanyan A,Sambandam V,Clayton MM,Choi SS,Xie GH,Diehl AM,et al. Hedgehog signaling blockade delays hepatocarcinogenesis induced by hepatitis B virus X protein.Cancer Res.2012;72:5912-20.
6. Zhang T,Zhang JP,You XN,Liu Q,Du YM,Gao YE,et al.Hepatitis B virus x protein modulates oncogene yes-associated protein by CREB to promote growth of hepatoma cells.Hepatology.2012;56:2051-59.
7. Geng X,Huang CH,Qin Y,McCombs JE,Yuan Q,Harry BL,et al.Hepatitis B virus X protein targets Bcl-2 proteins to increase intracellular calcium,required for virus replication and cell death induction.Proc Natl Acad Sci USA.2012;109:18471-6.
8. You XN,Liu FB,Zhang T,Li YH,Ye LH,Zhang XD.Hepatitis B virus X protein upregulates oncogene Rab18 to result in the dysregulation of lipogenesis and proliferatio n of hepatoma cells.Carcinogenesis.2013;34:1644-52.
9. Zhang S,Shan CL,Cui WJ,You XN,Du YM,Kong GY,et al.Hepatitis B virus X protein protects hepatoma and hepatic cells from complement-dependent cytotoxicity by up-regulation of CD46.FEBS Lett.2013;587:645-51.
10. Gao Y,Feng JY,Yang G,Zhang SQ,Liu YX,Bu YN,et al.Hepatitis B virus X protein-elevated MSL2 modulates hepatitis B virus covalently closed circular DNA by inducing degradation of APOBEC3B to enhance hepatocarcinogenesis.Hepatology.2017;66:1413-29.
11. Park ES,Park YK,Shin CY,Park SH,Ahn SH,Kim DH,et al.Hepatitis B virus inhibits liver regeneration via epigenetic regulation of urokinase-type plasminogen activator.Hepatology.2013;58:762-76.
12. Song K,Han C,Wu T.Epigenetic regulation of miR-122 expression by PPAR gamma/RXR alpha complex and HBx in hepatocellular carcinoma.Hepatology.2012;56:609a-10a.
13. Bartel DP.MicroRNAs:genomics,biogenesis,mechanism,and function.Cell.2004;116:281-97.
14. Soifer HS,Rossi JJ,Saetrom P.MicroRNAs in disease and potential therapeutic applications.Mol Ther.2007;15:2070-9.
15. Finnegan EF,Pasquinelli AE.MicroRNA biogenesis:regulating the regulators.Crit Rev Biochem Mol Biol.2013;48:51-68.
16. Gurtan AM,Sharp PA.The role of miRNAs in regulating gene expression networks.J Mol Biol.2013;425:3582-600.
17. Gregory RI,Chendrimada TP,Shiekhattar R.MicroRNA biogenesis:Isolation a nd characterization of the microprocessor complex.In:Ying SY.MicroRNA Protocols.Totowa,NJ:Humana Press;2006;33-47.
18. Jinek M,Doudna JA.A three-dimensional view of the molecular machinery of RNA interference.Nature.2009;457:405-12.
19. Chen RJ,Kelly G,Sengupta A,Heydendael W,Nicholas B,Beltrami S,et al.MicroRNAs as biomarkers of resilience or vulnerability to stress.Neuroscience.2015;305:36-48.
20. Haflidadottir BS,Ceder Y.Exosomal microRNAs as potential biomarkers in castration-resistant prostate cancer.Eur Urol.2015;67:42-3.
21. Kriegel AJ,Baker MA,Liu Y,Liu PY,Cowley AW Jr,Liang MY.Endogenous microRNAs in human microvascular endothelial cells regulate mRNAs encoded by hypertension-related genes.Hypertension.2015;66:793-9.
22. Wang YH,Chen J,Lin ZY,Cao J,Huang HL,Jiang Y,et al.Role of deregulated microRNAs in non-small cell lung cancer progression using fresh-frozen and formalin-fixed,paraffin-embedded samples.Oncol Lett.2016;11:801-8.
23. Chakravarthi BVSK,Pathi SS,Goswami MT,Cieslik M,Zheng H,Nallasivam S,et al.The miR-124-prolyl hydroxylase P4HA1-MMP1axis plays a critical role in prostate cancer progression.Oncotarget.2014;5:6654-69.
24. Tan Gana NH,Onuki T,Victoriano AFB,Okamoto T.MicroRNAs in HIV-1 infection:an integration of viral and cellular interaction at the genomic level.Front Microbiol.2012;3:306.
25. Drury RE,O'Connor D,Pollard AJ.The clinical application of microRNAs in infectious disease.Front Immunol.2017;8:1182.
26. Hou J,Wang P,Lin L,Liu XG,Ma F,An HZ,et al.MicroRNA-146a feedback inhibits RIG-1-dependent TypeⅠIFN production in macrophages by targeting TRAF6,IRAK1,and IRAK2.J Immunol.2009;183:2150-8.
27. Rossato M,Curtale G,Tamassia N,Castellucci M,Mori L,Gasperini S,et al.IL-10-induced micro RNA-187 negatively regulates TNF-α,IL-6,and IL-12p40 production in TLR4-stimulated monocytes.Proc Natl Acad Sci USA.2012;109:E3101-10.
28. Curtale G,Mirolo M,Renzi TA,Rossato M,Bazzoni F,Locati M.Negative regulation of Toll-like receptor 4 signaling by IL-10-dependent microRNA-146b.Proc Natl Acad Sci USA.2013;110:11499-504.
29. Lecellier CH,Dunoyer P,Arar K,Lehmann-Che J,Eyquem S,Himber C,et al.A cellular microRNA mediates antiviral defense in human cells.Science.2005;308:557-60.
30. Nathans R,Chu CY,Serquina AK,Lu CC,Cao H,Rana TM.Cellular microRNA and P bodies modulate host-HIV-1interactions.Mol Cell.2009;34:696-709.
31. Bai XT,Nicot C.miR-28-3p is a cellular restriction factor thatinhibits human T cell leukemia virus,type 1(HTLV-1)replication and virus infection.J Biol Chem.2015;290:5381-90.
32. Hu W,Wang XJ,Ding XR,Li Y,Zhang XJ,Xie PW,et al.MicroRNA-141 represses HBV replication by targeting PPARA.PLoS One.2012;7:e34165.
33. Jin J,Tang SH,Xia L,Du R,Xie HH,Song JG,et al.MicroRNA-501promotes HBV replication by targeting HBXIP.Biochem Biophys Res Commun.2013;430:1228-33.
34. Dar AA,Majid S,Rittsteuer C,de Semir D,Bezrookove V,Tong S,et al.The role of miR-18b in MDM2-p53 pathway signaling and melanoma progression.J Natl Cancer Inst.2013;105:433-42.
35. Wu JH,Wang YH,Wang W,Shen W,Sang YZ,Liu L,et al.MiR-18b suppresses high-glucose-induced proliferation in HRECs by targeting IGF-1/IGF1R signaling pathways.Int J Biochem Cell Biol.2016;73:41-52.
36. Lujambio A,Calin GA,Villanueva A,Ropero S,Sanchez-Cespedes M,Blanco D,et al.A microRNA DNA methylation signature for human cancer metastasis.Proc Natl Acad Sci USA.2008;105:13556-61.
37. Lopez-Serra P,Esteller M.DNA methylation-associated silencing of tumor-suppressor microRNAs in cancer.Oncogene.2012;31:1609-22.
38. Zhang T,Zhang JP,Cui M,Liu FB,You XN,Du YM,et al.Hepatitis B virus X protein inhibits tumor suppressor miR-205through inducing hypermethylation of miR-205 promoter to enhance carcinogenesis.Neoplasia.2013;15:1282-91.
39. Vanden Bosch A,Raemaekers T,Denayer S,Torrekens S,Smets N,Moermans K,et al.NuSAP is essential for chromatin-induced spindle formation during early embryogenesis.J Cell Sci.2010;123:3244-55.
40. Okamoto A,Higo M,Shiiba M,Nakashima D,Koyama T,Miyamoto I,et al.Down-regulation of nucleolar and spindleassociated protein 1(NUSAP1)expression suppresses tumor and cell proliferation and enhances anti-tumor effect of paclitaxel in oral squamous cell carcinoma.PLoS One.2015;10:e0142252.
41. Gordon CA,Gulzar ZG,Brooks JD.NUSAP1 expression is upregulated by loss of RBI in prostate cancer cells.Prostate.2015;75:517-26.
42. Chen LD,Zhuo DQ,Chen JK,Yuan HY.Screening feature genes of lung carcinoma with DNA micro array analysis.Int J Clin Exp Med.2015;8:12161-71.
43. Satow R,Shitashige M,Kanai Y,Takeshita F,Ojima H,Jigami T,et al.Combined functional genome survey of therapeutic targets for hepatocellular carcinoma.Clin Cancer Res.2010;16:2518-28.
44. Liu YW,Xia R,Lu K,Xie M,Yang F,Sun M,et al.LincRNAFEZFIAS1 represses p21 expression to promote gastric cancer proliferation th ro ugh LSD1-Mediated H3K4me2 deme thylation.Mol Cancer.2017;16:39.
45. Xu T,Zhu Y,Xiong YJ,Ge YY,Yun JP,Zhuang SM.MicroRNA-195 suppresses tumorigenicity and regulates G1/S transition of human hepatocellular carcinoma cells.Hepatology.2009;50:113-21.
46. Fang L,Zhang M,Chen L,Xiong H,Ge YK,Lu W,et al.Downregulation of nucleolar and spindle-associated protein 1expression suppresses cell migration,proliferation and invasion in renal cell carcinoma.Oncol Rep.2016;36:1506-16.
47. Bimonte S,Leongito M,Barbieri A,Del Vecchio V,Falco M,Giudice A,et al.The therapeutic targets of miRNA in hepatic cancer stem cells.Stem Cells Int.2016;2016:1065230.
48. Braconi C,Henry JC,Kogure T,Schmittgen T,Patel T.The role of MicroRNAs in human liver cancers.Semin Oncol.2011;38:752-63.
49. Coppola N,Potenza N,Pisaturo M,Mosca N,Tonziello G,Signoriello G,et al.Liver microRNA hsa-miR-125a-5p in HBV chronic infection:correlation with HBV replication and disease progression.PLoS One.2013;8:e65336.
50. Jiang RQ,Deng L,Zhao L,Li XC,Zhang F,Xia YX,et al.miR-22promotes HBV-related hepatocellular carcinoma development in males.Clin Cancer Res.2011;17:5593-603.
51. Khokhar A,Noorali S,Sheraz M,Mahalingham K,Pace DG,Khanani MR,et al.Computational analysis to predict functional role of hsa-miR-3065-3p as an antiviral therapeutic agent for treatment of triple infections:HCV,HIV-1,and HBV.Libyan J Med.2012;7:19774.
52. Li BB,Li DL,Chen C,Liu BH,Xia CY,Wu HJ,et al.Potentials of the elevated circulating miR-185 level as a biomarker for early diagnosis of HBV-related liver fibrosis.Sci Rep.2016;6:34157.
53. Katayama Y,Maeda M,Miyaguchi K,Nemoto S,Yasen M,TanakaS,et al.Identification of pathogenesis-related microRNAs in hepatocellular carcinoma by expression profiling.Oncol Lett.20124:817-23.
54. Zhang ZZ,Liu X,Wang DQ,Teng MK,Niu LW,Huang AL,et al.Hepatitis B virus and hepatocellular carcinoma at the miRNA level World J Gastroenterol.2011;17:3353-8.
55. Chen L,Yang L,Qiao F,Hu X,Li S,Yao L,et al.High levels of nucleolar spindle-associated protein and reduced levels of BRCA1expression predict poor prognosis in triple-negative breast cancer.PLoS One.2015;10:e0140572.
56. Wu XQ,Xu BK,Yang C,Wang WT,Zhong DQ,Zhao Z,et al.Nucleolar and spindle associated protein 1 promotes the aggressiveness of astrocytoma by activating the Hedgehog signaling pathway.J Exp Clin Cancer Res.2017;36:127.
57. Gordon CA,Gong X,Ganesh D,Brooks JD.NUSAP1 promotes invasion and metastasis of prostate cancer.Oncotarget.2017;8:29935-50.