沉默CIT基因可抑制前列腺癌细胞的增殖和转移（英文）
|
摘要
目的探究Citron Rho-interacting serine/threonine kinase(CIT)基因在前列腺癌中的表达特点,并探讨CIT基因对PC-3细胞增殖、迁移和侵袭能力的调控作用及其可能的分子机制。方法利用TCGA和MSKCC数据库分析CIT在前列腺癌组织的表达水平和临床相关性。采用siRNA干扰CIT的表达,采用CCK-8、划痕实验和Transwell实验检测CIT对前列腺癌PC-3细胞系的增殖、迁移和侵袭特性的影响。采用Western blotting检测CIT对上皮-间质转化和Hippo-YAP通路的调控作用。结果 TCGA数据库分析表明,CIT在前列腺癌组织中表达显著上调(P<0.05);MSKCC数据库分析表明,CIT表达水平与N分期(P<0.05)、M分期(P<0.001)、Gleason评分(P=0.010)和PSA水平(P=0.004)成正相关关系。PC-3细胞中沉默CIT表达可显著抑制细胞的增殖、迁移和侵袭,逆转上皮-间质转化进程,并抑制Hippo-YAP通路的激活。结论 CIT基因可能是前列腺癌的致病基因,其分子机制可能与Hippo-YAP通路的激活有关。
Objective Citron Rho-interacting serine/threonine kinase(CIT) was identified recently as an oncogene involved in the progression of various malignant tumors, but its role in prostate cancer(PCa) remains unclear. In this study, we aimed to investigate the biological functions of CIT in PCa. Methods We analyzed the expression of CIT in PCa tissues and its clinical correlations based on the Cancer Genome Atlas(TCGA) and Memorial Sloan-Kettering Cancer Center(MSKCC) dataset. We then examined the effects of RNA interference-mediated CIT silencing on the proliferation, migration and invasion of PC-3 cells using cell counting kit-8,wound healing assay and Transwell assay. We also investigated the effect of CIT silencing on epithelial-mesenchymal transition(EMT) and Hippo-Yap signaling pathway in the cells using Western blotting. Results CIT expression was significantly elevated in PCa tissues from TCGA cohort(P<0.05). MSKCC dataset analysis showed that an elevated expression of CIT was significantly correlated with N stage(P=0.001), distant metastasis(P<0.001), Gleason score(P=0.010) and PSA(P=0.004). In cultured PC-3 cells,knockdown of CIT significantly inhibited cell proliferation, migration and invasion, reversed the EMT phenotype and decreased the expression and activity of YAP. Conclusion CIT might function as an oncogene in PCa by modulating the Hippo-YAP signaling pathway and serve as a candidate therapeutic target for PCa.
Objective Citron Rho-interacting serine/threonine kinase(CIT) was identified recently as an oncogene involved in the progression of various malignant tumors, but its role in prostate cancer(PCa) remains unclear. In this study, we aimed to investigate the biological functions of CIT in PCa. Methods We analyzed the expression of CIT in PCa tissues and its clinical correlations based on the Cancer Genome Atlas(TCGA) and Memorial Sloan-Kettering Cancer Center(MSKCC) dataset. We then examined the effects of RNA interference-mediated CIT silencing on the proliferation, migration and invasion of PC-3 cells using cell counting kit-8,wound healing assay and Transwell assay. We also investigated the effect of CIT silencing on epithelial-mesenchymal transition(EMT) and Hippo-Yap signaling pathway in the cells using Western blotting. Results CIT expression was significantly elevated in PCa tissues from TCGA cohort(P<0.05). MSKCC dataset analysis showed that an elevated expression of CIT was significantly correlated with N stage(P=0.001), distant metastasis(P<0.001), Gleason score(P=0.010) and PSA(P=0.004). In cultured PC-3 cells,knockdown of CIT significantly inhibited cell proliferation, migration and invasion, reversed the EMT phenotype and decreased the expression and activity of YAP. Conclusion CIT might function as an oncogene in PCa by modulating the Hippo-YAP signaling pathway and serve as a candidate therapeutic target for PCa.
引文
[1]Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J]CA Cancer J Clin,2015,65(2):87-108.
[2]Kohli M,Tindall DJ.New developments in the medical management o prostate cancer[J].Mayo Clin Proc,2010,85(1):77-86.
[3]Carlin B I,Andriole G L.The natural history,skeletal complications and management of bone metastases in patients with prostate carcinoma[J].Cancer,2000,88(12 Suppl):2989-94.
[4]de Bono J S,Oudard S,Ozguroglu M,et al.Prednisone plus cabazitaxe or mitoxantrone for metastatic castration-resistant prostate cancer progressing after docetaxel treatment:a randomised open-label tria[J].Lancet,2010,376(9747):1147-54.
[5]Li H,Zhang Y,Zhang Y,et al.Rsf-1 overexpression in human prostate cancer,implication as a prognostic marker[J].Tumour Biol,2014,35(6):5771-6.
[6]Baca SC,Garraway LA.The genomic landscape of prostate cancer[J]Front Endocrinol(Lausanne),2012,3:69.
[7]Barbieri CE,Baca SC,Lawrence MS,et al.Exome sequencing identifies recurrent SPOP,FOXA1 and MED12 mutations in prostate cancer[J]Nat Genet,2012,44(6):685-9.
[8]Grasso CS,Wu YM,Robinson DR,et al.The mutational landscape o lethal castration-resistant prostate cancer[J].Nature,2012,487(7406):239-43.
[9]Barbieri CE,Bangma CH,Bjartell A,et al.The mutational landscape o prostate cancer[J].Eur Urol,2013,64(4):567-76.
[10]Wu Z,Zhu X,Xu W,et al.Up-regulation of CIT promotes the growth o colon cancer cells[J].Oncotarget,2017,8(42):71954-64.
[11]Bassi ZI,Audusseau M,Riparbelli MG,et al.Citron kinase controls a molecular network required for midbody formation in cytokinesis[J]Proc Natl Acad Sci USA,2013,110(24):9782-7.
[12]Bassi ZI,Verbrugghe KJ,Capalbo L,et al.Sticky/Citron kinase maintains proper RhoA localization at the cleavage site during cytokinesis[J].J Cell Biol,2011,195(4):595-603.
[13]Di Cunto F,Imarisio S,Hirsch E,et al.Defective neurogenesis in citron kinase knockout mice by altered cytokinesis and massive apoptosis[J]Neuron,2000,28(1):115-27.
[14]Fu Y,Huang J,Wang K S,et al.RNA interference targeting CITRONcan significantly inhibit the proliferation of hepatocellular carcinoma cells[J].Mol Biol Rep,2011,38(2):693-702.
[15]Whitworth H,Bhadel S,Ivey M,et al.Identification of kinases regulating prostate cancer cell growth using an RNAi phenotypic screen[J].PLoS One,2012,7(6):e38950.
[16]Ye G,Huang K,Yu J,et al.MicroRNA-647 targets SRF-MYH9 axis to suppress invasion and metastasis of gastric cancer[J].Theranostics,2017,7(13):3338-53.
[17]Gao J,Aksoy BA,Dogrusoz U,et al.Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal[J].Sci Signal,2013,6(269):pl1.
[18]Montanari M,Rossetti S,Cavaliere C,et al.Epithelial-mesenchymal transition in prostate cancer:an overview[J].Oncotarget,2017,8(21):35376-89.
[19]Lo UG,Lee CF,Lee MS,et al.The role and mechanism of epithelial-tomesenchymal transition in prostate cancer progression[J].Int J Mol Sci,2017,18(10).
[20]Maier J,Traenkle B,Rothbauer U.Visualizing epithelial-mesenchymal transition using the chromobody technology[J].Cancer Res,2016,76(19):5592-96.
[21]Harvey KF,Zhang X,Thomas DM.The Hippo pathway and human cancer[J].Nat Rev Cancer,2013,13(4):246-57.
[22]Zhao B,Tumaneng K,Guan KL.The Hippo pathway in organ size control,tissue regeneration and stem cell self-renewal[J].Nat Cel Biol,2011,13(8):877-83.
[23]Chan SW,Lim CJ,Loo LS,et al.TEADs mediate nuclear retention o TAZ to promote oncogenic transformation[J].J Biol Chem,2009,284(21):14347-58.
[24]Overholtzer M,Zhang J,Smolen GA,et al.Transforming properties o YAP,a candidate oncogene on the chromosome 11q22 amplicon[J]Proc Natl Acad Sci USA,2006,103(33):12405-10.
[25]Zhang H,Liu CY,Zha ZY,et al.TEAD transcription factors mediate the function of TAZ in cell growth and epithelial-mesenchyma transition[J].J Biol Chem,2009,284(20):13355-62.
[26]Zhang L,Yang S,Chen X,et al.The hippo pathway effector YAPregulates motility,invasion,and castration-resistant growth of prostate cancer cells[J].Mol Cell Biol,2015,35(8):1350-62.
[27]Collak FK,Demir U,Ozkanli S,et al.Increased expression of YAP1 in prostate cancer correlates with extraprostatic extension[J].Cancer Biol Med,2017,14(4):405-13.
[28]Fernandez-LA,Northcott PA,Dalton J,et al.YAP1 is amplified and up-regulated in hedgehog-associated medulloblastomas and mediates Sonic hedgehog-driven neural precursor proliferation[J].Genes Dev2009,23(23):2729-41.
[2]Kohli M,Tindall DJ.New developments in the medical management o prostate cancer[J].Mayo Clin Proc,2010,85(1):77-86.
[3]Carlin B I,Andriole G L.The natural history,skeletal complications and management of bone metastases in patients with prostate carcinoma[J].Cancer,2000,88(12 Suppl):2989-94.
[4]de Bono J S,Oudard S,Ozguroglu M,et al.Prednisone plus cabazitaxe or mitoxantrone for metastatic castration-resistant prostate cancer progressing after docetaxel treatment:a randomised open-label tria[J].Lancet,2010,376(9747):1147-54.
[5]Li H,Zhang Y,Zhang Y,et al.Rsf-1 overexpression in human prostate cancer,implication as a prognostic marker[J].Tumour Biol,2014,35(6):5771-6.
[6]Baca SC,Garraway LA.The genomic landscape of prostate cancer[J]Front Endocrinol(Lausanne),2012,3:69.
[7]Barbieri CE,Baca SC,Lawrence MS,et al.Exome sequencing identifies recurrent SPOP,FOXA1 and MED12 mutations in prostate cancer[J]Nat Genet,2012,44(6):685-9.
[8]Grasso CS,Wu YM,Robinson DR,et al.The mutational landscape o lethal castration-resistant prostate cancer[J].Nature,2012,487(7406):239-43.
[9]Barbieri CE,Bangma CH,Bjartell A,et al.The mutational landscape o prostate cancer[J].Eur Urol,2013,64(4):567-76.
[10]Wu Z,Zhu X,Xu W,et al.Up-regulation of CIT promotes the growth o colon cancer cells[J].Oncotarget,2017,8(42):71954-64.
[11]Bassi ZI,Audusseau M,Riparbelli MG,et al.Citron kinase controls a molecular network required for midbody formation in cytokinesis[J]Proc Natl Acad Sci USA,2013,110(24):9782-7.
[12]Bassi ZI,Verbrugghe KJ,Capalbo L,et al.Sticky/Citron kinase maintains proper RhoA localization at the cleavage site during cytokinesis[J].J Cell Biol,2011,195(4):595-603.
[13]Di Cunto F,Imarisio S,Hirsch E,et al.Defective neurogenesis in citron kinase knockout mice by altered cytokinesis and massive apoptosis[J]Neuron,2000,28(1):115-27.
[14]Fu Y,Huang J,Wang K S,et al.RNA interference targeting CITRONcan significantly inhibit the proliferation of hepatocellular carcinoma cells[J].Mol Biol Rep,2011,38(2):693-702.
[15]Whitworth H,Bhadel S,Ivey M,et al.Identification of kinases regulating prostate cancer cell growth using an RNAi phenotypic screen[J].PLoS One,2012,7(6):e38950.
[16]Ye G,Huang K,Yu J,et al.MicroRNA-647 targets SRF-MYH9 axis to suppress invasion and metastasis of gastric cancer[J].Theranostics,2017,7(13):3338-53.
[17]Gao J,Aksoy BA,Dogrusoz U,et al.Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal[J].Sci Signal,2013,6(269):pl1.
[18]Montanari M,Rossetti S,Cavaliere C,et al.Epithelial-mesenchymal transition in prostate cancer:an overview[J].Oncotarget,2017,8(21):35376-89.
[19]Lo UG,Lee CF,Lee MS,et al.The role and mechanism of epithelial-tomesenchymal transition in prostate cancer progression[J].Int J Mol Sci,2017,18(10).
[20]Maier J,Traenkle B,Rothbauer U.Visualizing epithelial-mesenchymal transition using the chromobody technology[J].Cancer Res,2016,76(19):5592-96.
[21]Harvey KF,Zhang X,Thomas DM.The Hippo pathway and human cancer[J].Nat Rev Cancer,2013,13(4):246-57.
[22]Zhao B,Tumaneng K,Guan KL.The Hippo pathway in organ size control,tissue regeneration and stem cell self-renewal[J].Nat Cel Biol,2011,13(8):877-83.
[23]Chan SW,Lim CJ,Loo LS,et al.TEADs mediate nuclear retention o TAZ to promote oncogenic transformation[J].J Biol Chem,2009,284(21):14347-58.
[24]Overholtzer M,Zhang J,Smolen GA,et al.Transforming properties o YAP,a candidate oncogene on the chromosome 11q22 amplicon[J]Proc Natl Acad Sci USA,2006,103(33):12405-10.
[25]Zhang H,Liu CY,Zha ZY,et al.TEAD transcription factors mediate the function of TAZ in cell growth and epithelial-mesenchyma transition[J].J Biol Chem,2009,284(20):13355-62.
[26]Zhang L,Yang S,Chen X,et al.The hippo pathway effector YAPregulates motility,invasion,and castration-resistant growth of prostate cancer cells[J].Mol Cell Biol,2015,35(8):1350-62.
[27]Collak FK,Demir U,Ozkanli S,et al.Increased expression of YAP1 in prostate cancer correlates with extraprostatic extension[J].Cancer Biol Med,2017,14(4):405-13.
[28]Fernandez-LA,Northcott PA,Dalton J,et al.YAP1 is amplified and up-regulated in hedgehog-associated medulloblastomas and mediates Sonic hedgehog-driven neural precursor proliferation[J].Genes Dev2009,23(23):2729-41.