Ubiquitin-specific protease 22 enhances intestinal cell proliferation and tissue regeneration after intestinal ischemia reperfusion injury
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摘要
BACKGROUND Intestinal ischemia reperfusion(I/R) injury is a serious but common pathophysiological process of many diseases, resulting in a high mortality rate in clinical practice. Ubiquitin-specific protease 22(USP22) acts as regulator of cell cycle progression, proliferation, and tumor invasion. Depleted USP22 expression has been reported to contribute to arrested cell cycle and disrupted generation of differentiated cell types in crypts and villi. However, the role of USP22 in intestinal damage recovery has not been investigated. Therefore, elucidation of the underlying mechanism of USP22 in intestinal I/R injury may help to improve the tissue repair and patient prognosis in clinical practice.AIM To investigate the role of USP22 in intestinal cell proliferation and regeneration after intestinal I/R injury.METHODS An animal model of intestinal I/R injury was generated in male Sprague-Dawley rats by occlusion of the superior mesenteric artery followed by reperfusion.Chiu's scoring system was used to grade the damage to the intestinal mucosa. An in vitro model was developed by incubating rat intestinal epithelial IEC-6 cells in hypoxia/reoxygenation conditions in order to simulate I/R in vivo. siRNA and overexpression plasmid were used to regulate the expression of USP22. USP22,Cyclin D1, and proliferating cell nuclear antigen(PCNA) expression levels were measured by Western blot analysis and immunohistochemistry staining. Cell survival(viability) and cell cycle were evaluated using the Cell Counting Kit-8and flow cytometry, respectively.RESULTS USP22 expression was positively correlated with the expression levels of PCNA and Cyclin D1 both in vivo and in vitro, which confirmed that USP22 was involved in cell proliferation and intestinal regeneration after intestinal I/R injury. Decreased levels of Cyclin D1 and cell cycle arrest were observed in the USP22 knockdown group(P < 0.05), while opposite results were observed in the USP22 overexpression group(P < 0.05). In addition, increased expression of USP22 was related to improved intestinal pathology or IEC-6 cell viability after I/R or hypoxia/reoxygenation. These results suggested that USP22 may exert a protective effect on intestinal I/R injury by regulating cell proliferation and facilitating tissue regeneration.CONCLUSION USP22 is correlated with promoting intestinal cell proliferation and accelerating intestinal tissue regeneration after intestinal I/R injury and may serve as a potential target for therapeutic development for tissue repair during intestinal I/R injury.
BACKGROUND Intestinal ischemia reperfusion(I/R) injury is a serious but common pathophysiological process of many diseases, resulting in a high mortality rate in clinical practice. Ubiquitin-specific protease 22(USP22) acts as regulator of cell cycle progression, proliferation, and tumor invasion. Depleted USP22 expression has been reported to contribute to arrested cell cycle and disrupted generation of differentiated cell types in crypts and villi. However, the role of USP22 in intestinal damage recovery has not been investigated. Therefore, elucidation of the underlying mechanism of USP22 in intestinal I/R injury may help to improve the tissue repair and patient prognosis in clinical practice.AIM To investigate the role of USP22 in intestinal cell proliferation and regeneration after intestinal I/R injury.METHODS An animal model of intestinal I/R injury was generated in male Sprague-Dawley rats by occlusion of the superior mesenteric artery followed by reperfusion.Chiu's scoring system was used to grade the damage to the intestinal mucosa. An in vitro model was developed by incubating rat intestinal epithelial IEC-6 cells in hypoxia/reoxygenation conditions in order to simulate I/R in vivo. siRNA and overexpression plasmid were used to regulate the expression of USP22. USP22,Cyclin D1, and proliferating cell nuclear antigen(PCNA) expression levels were measured by Western blot analysis and immunohistochemistry staining. Cell survival(viability) and cell cycle were evaluated using the Cell Counting Kit-8and flow cytometry, respectively.RESULTS USP22 expression was positively correlated with the expression levels of PCNA and Cyclin D1 both in vivo and in vitro, which confirmed that USP22 was involved in cell proliferation and intestinal regeneration after intestinal I/R injury. Decreased levels of Cyclin D1 and cell cycle arrest were observed in the USP22 knockdown group(P < 0.05), while opposite results were observed in the USP22 overexpression group(P < 0.05). In addition, increased expression of USP22 was related to improved intestinal pathology or IEC-6 cell viability after I/R or hypoxia/reoxygenation. These results suggested that USP22 may exert a protective effect on intestinal I/R injury by regulating cell proliferation and facilitating tissue regeneration.CONCLUSION USP22 is correlated with promoting intestinal cell proliferation and accelerating intestinal tissue regeneration after intestinal I/R injury and may serve as a potential target for therapeutic development for tissue repair during intestinal I/R injury.
BACKGROUND Intestinal ischemia reperfusion(I/R) injury is a serious but common pathophysiological process of many diseases, resulting in a high mortality rate in clinical practice. Ubiquitin-specific protease 22(USP22) acts as regulator of cell cycle progression, proliferation, and tumor invasion. Depleted USP22 expression has been reported to contribute to arrested cell cycle and disrupted generation of differentiated cell types in crypts and villi. However, the role of USP22 in intestinal damage recovery has not been investigated. Therefore, elucidation of the underlying mechanism of USP22 in intestinal I/R injury may help to improve the tissue repair and patient prognosis in clinical practice.AIM To investigate the role of USP22 in intestinal cell proliferation and regeneration after intestinal I/R injury.METHODS An animal model of intestinal I/R injury was generated in male Sprague-Dawley rats by occlusion of the superior mesenteric artery followed by reperfusion.Chiu's scoring system was used to grade the damage to the intestinal mucosa. An in vitro model was developed by incubating rat intestinal epithelial IEC-6 cells in hypoxia/reoxygenation conditions in order to simulate I/R in vivo. siRNA and overexpression plasmid were used to regulate the expression of USP22. USP22,Cyclin D1, and proliferating cell nuclear antigen(PCNA) expression levels were measured by Western blot analysis and immunohistochemistry staining. Cell survival(viability) and cell cycle were evaluated using the Cell Counting Kit-8and flow cytometry, respectively.RESULTS USP22 expression was positively correlated with the expression levels of PCNA and Cyclin D1 both in vivo and in vitro, which confirmed that USP22 was involved in cell proliferation and intestinal regeneration after intestinal I/R injury. Decreased levels of Cyclin D1 and cell cycle arrest were observed in the USP22 knockdown group(P < 0.05), while opposite results were observed in the USP22 overexpression group(P < 0.05). In addition, increased expression of USP22 was related to improved intestinal pathology or IEC-6 cell viability after I/R or hypoxia/reoxygenation. These results suggested that USP22 may exert a protective effect on intestinal I/R injury by regulating cell proliferation and facilitating tissue regeneration.CONCLUSION USP22 is correlated with promoting intestinal cell proliferation and accelerating intestinal tissue regeneration after intestinal I/R injury and may serve as a potential target for therapeutic development for tissue repair during intestinal I/R injury.
引文
1 Collard CD,Gelman S.Pathophysiology,clinical manifestations,and prevention of ischemia-reperfusion injury.Anesthesiology 2001;94:1133-1138[PMID:11465607 DOI:10.1097/00000542-200106000-00030]
2 Tendler DA.Acute intestinal ischemia and infarction.Semin Gastrointest Dis 2003;14:66-76[PMID:12889581 DOI:10.1016/j.crma.2009.11.008]
3 Blikslager AT,Moeser AJ,Gookin JL,Jones SL,Odle J.Restoration of barrier function in injured intestinal mucosa.Physiol Rev 2007;87:545-564[PMID:17429041 DOI:10.1152/physrev.00012.2006]
4 Zhou W,Yao J,Wang G,Chen Z,Li Z,Feng D,Li Y,Qasim W,Tan W,Ning S,Tian X.PKCζphosphorylates TRAF2 to protect against intestinal ischemia-reperfusion-induced injury.Cell Death Dis2017;8:e2935[PMID:28726782 DOI:10.1038/cddis.2017.310]
5 Eltzschig HK,Eckle T.Ischemia and reperfusion--from mechanism to translation.Nat Med 2011;17:1391-1401[PMID:22064429 DOI:10.1038/nm.2507]
6 Higuchi S,Wu R,Zhou M,Marini CP,Ravikumar TS,Wang P.Gut hyperpermiability after ischemia and reperfusion:Attenuation with adrenomedullin and its binding protein treatment.Int J Clin Exp Pathol2008;1:409-418[PMID:18787625 DOI:10.1080/14992020802286202]
7 Zu G,Yao J,Ji A,Ning S,Luo F,Li Z,Feng D,Rui Y,Li Y,Wang G,Tian X.Nurr1 promotes intestinal regeneration after ischemia/reperfusion injury by inhibiting the expression of p21(Waf1/Cip1).J Mol Med(Berl)2017;95:83-95[PMID:27553040 DOI:10.1007/s00109-016-1464-6]
8 Grootjans J,Thuijls G,Derikx JP,van Dam RM,Dejong CH,Buurman WA.Rapid lamina propria retraction and zipper-like constriction of the epithelium preserves the epithelial lining in human small intestine exposed to ischaemia-reperfusion.J Pathol 2011;224:411-419[PMID:21547908 DOI:10.1002/path.2882]
9 Podolsky DK.Mucosal immunity and inflammation.V.Innate mechanisms of mucosal defense and repair:The best offense is a good defense.Am J Physiol 1999;277:G495-G499[PMID:10484372 DOI:10.1152/ajpgi.1999.277.3.G495]
10 Itoh H,Yagi M,Hasebe K,Fushida S,Tani T,Hashimoto T,Shimizu K,Miwa K.Regeneration of small intestinal mucosa after acute ischemia-reperfusion injury.Dig Dis Sci 2002;47:2704-2710[PMID:12498289 DOI:10.1023/A:1021049004188]
11 Mallick IH,Yang W,Winslet MC,Seifalian AM.Ischemia-reperfusion injury of the intestine and protective strategies against injury.Dig Dis Sci 2004;49:1359-1377[PMID:15481305 DOI:10.1093/rpd/ncm395]
12 Hart ML,Grenz A,Gorzolla IC,Schittenhelm J,Dalton JH,Eltzschig HK.Hypoxia-inducible factor-1α-dependent protection from intestinal ischemia/reperfusion injury involves ecto-5'-nucleotidase(CD73)and the A2B adenosine receptor.J Immunol 2011;186:4367-4374[PMID:21357264 DOI:10.4049/jimmunol.0903617]
13 Koutelou E,Hirsch CL,Dent SY.Multiple faces of the SAGA complex.Curr Opin Cell Biol 2010;22:374-382[PMID:20363118 DOI:10.1016/j.ceb.2010.03.005]
14 Atanassov BS,Dent SY.USP22 regulates cell proliferation by deubiquitinating the transcriptional regulator FBP1.EMBO Rep 2011;12:924-930[PMID:21779003 DOI:10.1038/embor.2011.140]
15 Zhang XY,Pfeiffer HK,Thorne AW,McMahon SB.USP22,an hSAGA subunit and potential cancer stem cell marker,reverses the polycomb-catalyzed ubiquitylation of histone H2A.Cell Cycle 2008;7:1522-1524[PMID:18469533 DOI:10.4161/cc.7.11.5962]
16 Wang L,Dent SY.Functions of SAGA in development and disease.Epigenomics 2014;6:329-339[PMID:25111486 DOI:10.2217/epi.14.22]
17 Stine ZE,Walton ZE,Altman BJ,Hsieh AL,Dang CV.MYC,Metabolism,and Cancer.Cancer Discov2015;5:1024-1039[PMID:26382145 DOI:10.1158/2159-8290.CD-15-0507]
18 Kim D,Hong A,Park HI,Shin WH,Yoo L,Jeon SJ,Chung KC.Deubiquitinating enzyme USP22positively regulates c-Myc stability and tumorigenic activity in mammalian and breast cancer cells.J Cell Physiol 2017;232:3664-3676[PMID:28160502 DOI:10.1002/jcp.25841]
19 Zhang XY,Varthi M,Sykes SM,Phillips C,Warzecha C,Zhu W,Wyce A,Thorne AW,Berger SL,McMahon SB.The putative cancer stem cell marker USP22 is a subunit of the human SAGA complex required for activated transcription and cell-cycle progression.Mol Cell 2008;29:102-111[PMID:18206973 DOI:10.1016/j.molcel.2007.12.015]
20 Benetatos L,Vartholomatos G,Hatzimichael E.Polycomb group proteins and MYC:The cancer connection.Cell Mol Life Sci 2014;71:257-269[PMID:23897499 DOI:10.1007/s00018-013-1426-x]
21 Kosinsky RL,Wegwitz F,Hellbach N,Dobbelstein M,Mansouri A,Vogel T,Begus-Nahrmann Y,Johnsen SA.Usp22 deficiency impairs intestinal epithelial lineage specification in vivo.Oncotarget 2015;6:37906-37918[PMID:26431380 DOI:10.18632/oncotarget.5412]
22 Kim J,Seo BS.How to calculate sample size and why.Clin Orthop Surg 2013;5:235-242[PMID:24009911 DOI:10.4055/cios.2013.5.3.235]
23 Faul F,Erdfelder E,Buchner A,Lang AG.Statistical power analyses using G*Power 3.1:Tests for correlation and regression analyses.Behav Res Methods 2009;41:1149-1160[PMID:19897823 DOI:10.3758/BRM.41.4.1149]
24 Faul F,Erdfelder E,Lang AG,Buchner A.G*Power 3:A flexible statistical power analysis program for the social,behavioral,and biomedical sciences.Behav Res Methods 2007;39:175-191[PMID:17695343]
25 Megison SM,Horton JW,Chao H,Walker PB.A new model for intestinal ischemia in the rat.J Surg Res1990;49:168-173[PMID:2381206 DOI:10.1016/0022-4804(90)90257-3]
26 Chiu CJ,McArdle AH,Brown R,Scott HJ,Gurd FN.Intestinal mucosal lesion in low-flow states.I.Amorphological,hemodynamic,and metabolic reappraisal.Arch Surg 1970;101:478-483[PMID:5457245DOI:10.1001/archsurg.1970.01340280030009]
27 Lee KK,Florens L,Swanson SK,Washburn MP,Workman JL.The deubiquitylation activity of Ubp8 is dependent upon Sgf11 and its association with the SAGA complex.Mol Cell Biol 2005;25:1173-1182[PMID:15657442 DOI:10.1128/mcb.25.3.1173-1182.2005]
28 Andika IB,Jamal A,Kondo H,Suzuki N.SAGA complex mediates the transcriptional up-regulation of antiviral RNA silencing.Proc Natl Acad Sci U S A 2017;114:E3499-E3506[PMID:28400515 DOI:10.1073/pnas.1701196114]
29 Li L,Osdal T,Ho Y,Chun S,McDonald T,Agarwal P,Lin A,Chu S,Qi J,Li L,Hsieh YT,Dos Santos C,Yuan H,Ha TQ,Popa M,Hovland R,Bruserud?,Gjertsen BT,Kuo YH,Chen W,Lain S,McCormack E,Bhatia R.SIRT1 activation by a c-MYC oncogenic network promotes the maintenance and drug resistance of human FLT3-ITD acute myeloid leukemia stem cells.Cell Stem Cell 2014;15:431-446[PMID:25280219 DOI:10.1016/j.stem.2014.08.001]
30 Lin Z,Yang H,Kong Q,Li J,Lee SM,Gao B,Dong H,Wei J,Song J,Zhang DD,Fang D.USP22antagonizes p53 transcriptional activation by deubiquitinating Sirt1 to suppress cell apoptosis and is required for mouse embryonic development.Mol Cell 2012;46:484-494[PMID:22542455 DOI:10.1016/j.molcel.2012.03.024]
31 Pelaseyed T,Bergstr?m JH,Gustafsson JK,Ermund A,Birchenough GM,Schütte A,van der Post S,Svensson F,Rodríguez-Pi?eiro AM,Nystr?m EE,Wising C,Johansson ME,Hansson GC.The mucus and mucins of the goblet cells and enterocytes provide the first defense line of the gastrointestinal tract and interact with the immune system.Immunol Rev 2014;260:8-20[PMID:24942678 DOI:10.1111/imr.12182]
32 Birchenough GM,Johansson ME,Gustafsson JK,Bergstr?m JH,Hansson GC.New developments in goblet cell mucus secretion and function.Mucosal Immunol 2015;8:712-719[PMID:25872481 DOI:10.1038/mi.2015.32]
33 Takasaki Y,Deng JS,Tan EM.A nuclear antigen associated with cell proliferation and blast transformation.J Exp Med 1981;154:1899-1909[PMID:6172535 DOI:10.1084/jem.154.6.1899]
34 Baldin V,Lukas J,Marcote MJ,Pagano M,Draetta G.Cyclin D1 is a nuclear protein required for cell cycle progression in G1.Genes Dev 1993;7:812-821[PMID:8491378 DOI:10.1101/gad.7.5.812]
35 Tam SW,Theodoras AM,Shay JW,Draetta GF,Pagano M.Differential expression and regulation of Cyclin D1 protein in normal and tumor human cells:Association with Cdk4 is required for Cyclin D1function in G1 progression.Oncogene 1994;9:2663-2674[PMID:8058330 DOI:10.1093/nar/22.17.3663]
36 Liu L,Zhang H,Shi L,Zhang W,Yuan J,Chen X,Liu J,Zhang Y,Wang Z.Inhibition of Rac1 activity induces G1/S phase arrest through the GSK3/cyclin D1 pathway in human cancer cells.Oncol Rep 2014;32:1395-1400[PMID:25109327 DOI:10.3892/or.2014.3388]
37 Gennaro VJ,Stanek TJ,Peck AR,Sun Y,Wang F,Qie S,Knudsen KE,Rui H,Butt T,Diehl JA,McMahon SB.Control of CCND1 ubiquitylation by the catalytic SAGA subunit USP22 is essential for cell cycle progression through G1 in cancer cells.Proc Natl Acad Sci U S A 2018;115:E9298-E9307[PMID:30224477 DOI:10.1073/pnas.1807704115]
38 Jiang S,Song C,Gu X,Wang M,Miao D,Lv J,Liu Y.Ubiquitin-Specific Peptidase 22 Contributes to Colorectal Cancer Stemness and Chemoresistance via Wnt/β-Catenin Pathway.Cell Physiol Biochem2018;46:1412-1422[PMID:29689565 DOI:10.1159/000489156]
39 Jiang S,Miao D,Wang M,Lv J,Wang Y,Tong J.MiR-30-5p suppresses cell chemoresistance and stemness in colorectal cancer through USP22/Wnt/β-catenin signaling axis.J Cell Mol Med 2019;23:630-640[PMID:30338942 DOI:10.1111/jcmm.13968]
40 Ma J,Rubin BK,Voynow JA.Mucins,Mucus,and Goblet Cells.Chest 2018;154:169-176[PMID:29170036 DOI:10.1016/j.chest.2017.11.008]
41 Johansson ME,Hansson GC.Immunological aspects of intestinal mucus and mucins.Nat Rev Immunol2016;16:639-649[PMID:27498766 DOI:10.1038/nri.2016.88]
2 Tendler DA.Acute intestinal ischemia and infarction.Semin Gastrointest Dis 2003;14:66-76[PMID:12889581 DOI:10.1016/j.crma.2009.11.008]
3 Blikslager AT,Moeser AJ,Gookin JL,Jones SL,Odle J.Restoration of barrier function in injured intestinal mucosa.Physiol Rev 2007;87:545-564[PMID:17429041 DOI:10.1152/physrev.00012.2006]
4 Zhou W,Yao J,Wang G,Chen Z,Li Z,Feng D,Li Y,Qasim W,Tan W,Ning S,Tian X.PKCζphosphorylates TRAF2 to protect against intestinal ischemia-reperfusion-induced injury.Cell Death Dis2017;8:e2935[PMID:28726782 DOI:10.1038/cddis.2017.310]
5 Eltzschig HK,Eckle T.Ischemia and reperfusion--from mechanism to translation.Nat Med 2011;17:1391-1401[PMID:22064429 DOI:10.1038/nm.2507]
6 Higuchi S,Wu R,Zhou M,Marini CP,Ravikumar TS,Wang P.Gut hyperpermiability after ischemia and reperfusion:Attenuation with adrenomedullin and its binding protein treatment.Int J Clin Exp Pathol2008;1:409-418[PMID:18787625 DOI:10.1080/14992020802286202]
7 Zu G,Yao J,Ji A,Ning S,Luo F,Li Z,Feng D,Rui Y,Li Y,Wang G,Tian X.Nurr1 promotes intestinal regeneration after ischemia/reperfusion injury by inhibiting the expression of p21(Waf1/Cip1).J Mol Med(Berl)2017;95:83-95[PMID:27553040 DOI:10.1007/s00109-016-1464-6]
8 Grootjans J,Thuijls G,Derikx JP,van Dam RM,Dejong CH,Buurman WA.Rapid lamina propria retraction and zipper-like constriction of the epithelium preserves the epithelial lining in human small intestine exposed to ischaemia-reperfusion.J Pathol 2011;224:411-419[PMID:21547908 DOI:10.1002/path.2882]
9 Podolsky DK.Mucosal immunity and inflammation.V.Innate mechanisms of mucosal defense and repair:The best offense is a good defense.Am J Physiol 1999;277:G495-G499[PMID:10484372 DOI:10.1152/ajpgi.1999.277.3.G495]
10 Itoh H,Yagi M,Hasebe K,Fushida S,Tani T,Hashimoto T,Shimizu K,Miwa K.Regeneration of small intestinal mucosa after acute ischemia-reperfusion injury.Dig Dis Sci 2002;47:2704-2710[PMID:12498289 DOI:10.1023/A:1021049004188]
11 Mallick IH,Yang W,Winslet MC,Seifalian AM.Ischemia-reperfusion injury of the intestine and protective strategies against injury.Dig Dis Sci 2004;49:1359-1377[PMID:15481305 DOI:10.1093/rpd/ncm395]
12 Hart ML,Grenz A,Gorzolla IC,Schittenhelm J,Dalton JH,Eltzschig HK.Hypoxia-inducible factor-1α-dependent protection from intestinal ischemia/reperfusion injury involves ecto-5'-nucleotidase(CD73)and the A2B adenosine receptor.J Immunol 2011;186:4367-4374[PMID:21357264 DOI:10.4049/jimmunol.0903617]
13 Koutelou E,Hirsch CL,Dent SY.Multiple faces of the SAGA complex.Curr Opin Cell Biol 2010;22:374-382[PMID:20363118 DOI:10.1016/j.ceb.2010.03.005]
14 Atanassov BS,Dent SY.USP22 regulates cell proliferation by deubiquitinating the transcriptional regulator FBP1.EMBO Rep 2011;12:924-930[PMID:21779003 DOI:10.1038/embor.2011.140]
15 Zhang XY,Pfeiffer HK,Thorne AW,McMahon SB.USP22,an hSAGA subunit and potential cancer stem cell marker,reverses the polycomb-catalyzed ubiquitylation of histone H2A.Cell Cycle 2008;7:1522-1524[PMID:18469533 DOI:10.4161/cc.7.11.5962]
16 Wang L,Dent SY.Functions of SAGA in development and disease.Epigenomics 2014;6:329-339[PMID:25111486 DOI:10.2217/epi.14.22]
17 Stine ZE,Walton ZE,Altman BJ,Hsieh AL,Dang CV.MYC,Metabolism,and Cancer.Cancer Discov2015;5:1024-1039[PMID:26382145 DOI:10.1158/2159-8290.CD-15-0507]
18 Kim D,Hong A,Park HI,Shin WH,Yoo L,Jeon SJ,Chung KC.Deubiquitinating enzyme USP22positively regulates c-Myc stability and tumorigenic activity in mammalian and breast cancer cells.J Cell Physiol 2017;232:3664-3676[PMID:28160502 DOI:10.1002/jcp.25841]
19 Zhang XY,Varthi M,Sykes SM,Phillips C,Warzecha C,Zhu W,Wyce A,Thorne AW,Berger SL,McMahon SB.The putative cancer stem cell marker USP22 is a subunit of the human SAGA complex required for activated transcription and cell-cycle progression.Mol Cell 2008;29:102-111[PMID:18206973 DOI:10.1016/j.molcel.2007.12.015]
20 Benetatos L,Vartholomatos G,Hatzimichael E.Polycomb group proteins and MYC:The cancer connection.Cell Mol Life Sci 2014;71:257-269[PMID:23897499 DOI:10.1007/s00018-013-1426-x]
21 Kosinsky RL,Wegwitz F,Hellbach N,Dobbelstein M,Mansouri A,Vogel T,Begus-Nahrmann Y,Johnsen SA.Usp22 deficiency impairs intestinal epithelial lineage specification in vivo.Oncotarget 2015;6:37906-37918[PMID:26431380 DOI:10.18632/oncotarget.5412]
22 Kim J,Seo BS.How to calculate sample size and why.Clin Orthop Surg 2013;5:235-242[PMID:24009911 DOI:10.4055/cios.2013.5.3.235]
23 Faul F,Erdfelder E,Buchner A,Lang AG.Statistical power analyses using G*Power 3.1:Tests for correlation and regression analyses.Behav Res Methods 2009;41:1149-1160[PMID:19897823 DOI:10.3758/BRM.41.4.1149]
24 Faul F,Erdfelder E,Lang AG,Buchner A.G*Power 3:A flexible statistical power analysis program for the social,behavioral,and biomedical sciences.Behav Res Methods 2007;39:175-191[PMID:17695343]
25 Megison SM,Horton JW,Chao H,Walker PB.A new model for intestinal ischemia in the rat.J Surg Res1990;49:168-173[PMID:2381206 DOI:10.1016/0022-4804(90)90257-3]
26 Chiu CJ,McArdle AH,Brown R,Scott HJ,Gurd FN.Intestinal mucosal lesion in low-flow states.I.Amorphological,hemodynamic,and metabolic reappraisal.Arch Surg 1970;101:478-483[PMID:5457245DOI:10.1001/archsurg.1970.01340280030009]
27 Lee KK,Florens L,Swanson SK,Washburn MP,Workman JL.The deubiquitylation activity of Ubp8 is dependent upon Sgf11 and its association with the SAGA complex.Mol Cell Biol 2005;25:1173-1182[PMID:15657442 DOI:10.1128/mcb.25.3.1173-1182.2005]
28 Andika IB,Jamal A,Kondo H,Suzuki N.SAGA complex mediates the transcriptional up-regulation of antiviral RNA silencing.Proc Natl Acad Sci U S A 2017;114:E3499-E3506[PMID:28400515 DOI:10.1073/pnas.1701196114]
29 Li L,Osdal T,Ho Y,Chun S,McDonald T,Agarwal P,Lin A,Chu S,Qi J,Li L,Hsieh YT,Dos Santos C,Yuan H,Ha TQ,Popa M,Hovland R,Bruserud?,Gjertsen BT,Kuo YH,Chen W,Lain S,McCormack E,Bhatia R.SIRT1 activation by a c-MYC oncogenic network promotes the maintenance and drug resistance of human FLT3-ITD acute myeloid leukemia stem cells.Cell Stem Cell 2014;15:431-446[PMID:25280219 DOI:10.1016/j.stem.2014.08.001]
30 Lin Z,Yang H,Kong Q,Li J,Lee SM,Gao B,Dong H,Wei J,Song J,Zhang DD,Fang D.USP22antagonizes p53 transcriptional activation by deubiquitinating Sirt1 to suppress cell apoptosis and is required for mouse embryonic development.Mol Cell 2012;46:484-494[PMID:22542455 DOI:10.1016/j.molcel.2012.03.024]
31 Pelaseyed T,Bergstr?m JH,Gustafsson JK,Ermund A,Birchenough GM,Schütte A,van der Post S,Svensson F,Rodríguez-Pi?eiro AM,Nystr?m EE,Wising C,Johansson ME,Hansson GC.The mucus and mucins of the goblet cells and enterocytes provide the first defense line of the gastrointestinal tract and interact with the immune system.Immunol Rev 2014;260:8-20[PMID:24942678 DOI:10.1111/imr.12182]
32 Birchenough GM,Johansson ME,Gustafsson JK,Bergstr?m JH,Hansson GC.New developments in goblet cell mucus secretion and function.Mucosal Immunol 2015;8:712-719[PMID:25872481 DOI:10.1038/mi.2015.32]
33 Takasaki Y,Deng JS,Tan EM.A nuclear antigen associated with cell proliferation and blast transformation.J Exp Med 1981;154:1899-1909[PMID:6172535 DOI:10.1084/jem.154.6.1899]
34 Baldin V,Lukas J,Marcote MJ,Pagano M,Draetta G.Cyclin D1 is a nuclear protein required for cell cycle progression in G1.Genes Dev 1993;7:812-821[PMID:8491378 DOI:10.1101/gad.7.5.812]
35 Tam SW,Theodoras AM,Shay JW,Draetta GF,Pagano M.Differential expression and regulation of Cyclin D1 protein in normal and tumor human cells:Association with Cdk4 is required for Cyclin D1function in G1 progression.Oncogene 1994;9:2663-2674[PMID:8058330 DOI:10.1093/nar/22.17.3663]
36 Liu L,Zhang H,Shi L,Zhang W,Yuan J,Chen X,Liu J,Zhang Y,Wang Z.Inhibition of Rac1 activity induces G1/S phase arrest through the GSK3/cyclin D1 pathway in human cancer cells.Oncol Rep 2014;32:1395-1400[PMID:25109327 DOI:10.3892/or.2014.3388]
37 Gennaro VJ,Stanek TJ,Peck AR,Sun Y,Wang F,Qie S,Knudsen KE,Rui H,Butt T,Diehl JA,McMahon SB.Control of CCND1 ubiquitylation by the catalytic SAGA subunit USP22 is essential for cell cycle progression through G1 in cancer cells.Proc Natl Acad Sci U S A 2018;115:E9298-E9307[PMID:30224477 DOI:10.1073/pnas.1807704115]
38 Jiang S,Song C,Gu X,Wang M,Miao D,Lv J,Liu Y.Ubiquitin-Specific Peptidase 22 Contributes to Colorectal Cancer Stemness and Chemoresistance via Wnt/β-Catenin Pathway.Cell Physiol Biochem2018;46:1412-1422[PMID:29689565 DOI:10.1159/000489156]
39 Jiang S,Miao D,Wang M,Lv J,Wang Y,Tong J.MiR-30-5p suppresses cell chemoresistance and stemness in colorectal cancer through USP22/Wnt/β-catenin signaling axis.J Cell Mol Med 2019;23:630-640[PMID:30338942 DOI:10.1111/jcmm.13968]
40 Ma J,Rubin BK,Voynow JA.Mucins,Mucus,and Goblet Cells.Chest 2018;154:169-176[PMID:29170036 DOI:10.1016/j.chest.2017.11.008]
41 Johansson ME,Hansson GC.Immunological aspects of intestinal mucus and mucins.Nat Rev Immunol2016;16:639-649[PMID:27498766 DOI:10.1038/nri.2016.88]