Septin2对大鼠肾小球系膜细胞增殖的影响
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摘要
目的：探讨Septin2基因表达变化对大鼠肾小球系膜细胞增殖的影响。
方法：培养原代大鼠肾小球系膜细胞,进行以下实验：1)将绿色荧光蛋白表达质粒pEGFP-C1用电穿孔转染的方法导入细胞,24h后荧光显微镜下观察绿色荧光蛋白表达情况。2)质粒电穿孔转染：细胞分组：①pRK5空质粒转染组,②pRK5-Septin2质粒转染组,分别进行电穿孔转染,Western blot检测两组细胞Septin2蛋白质表达水平,流式细胞仪分析Septin2对细胞周期进程的影响,Western blot检测系膜细胞周期调节蛋白(cyclin D1、cyclinE、p21)的表达变化。3)设计合成GAPDH的siRNA,依不同浓度分为①200nmol／L,②400nmol／L,③800nmol／L组,④无关对照转染组(siCon),电穿孔转染后培养48h,Western blot检测GAPDH蛋白质表达变化。4)siRNA电穿孔转染：细胞分组：①无关对照转染组(Sicon);②siSeptin2转染组。收集细胞蛋白,Western blot检测两组细胞Septin2蛋白质表达水平,流式细胞仪分析siSeptin2对细胞周期进程的影响。
结果：1)在80μg质粒,340V电压、550μF电容的电穿孔条件下,原代大鼠肾小球系膜细胞的电转染效率可以达到50%以上；2)与pRK5空质粒转染组相比,pRK5-Septin2质粒可使系膜细胞Septin2的蛋白质表达水平明显提高(P<0.05),流式细胞仪检测发现pRK5-Septin2质粒使G0／G1期细胞百分比明显增多而S期细胞百分比显著减少(P<0.05),同时细胞周期调节蛋白cyclin D1、cyclinE表达水平下调,p21的表达水平上调(P<0.05)；3) siGAPDH浓度为800nmol／L时蛋白质水平抑制效果最为显著(P<0.05)；4)与无关对照转染组(siCon)比较,siSeptin2转染组Septin2蛋白质表达水平明显降低,流式细胞仪检测结果显示两组细胞G0／G1期及S期细胞百分比无明显差异。
结论：Septin2过表达可使大鼠肾小球系膜细胞由G0／G1期向S期转化的百分比减少,细胞周期正调控蛋白表达减少而负调控蛋白表达增多,提示可能通过调节细胞周期蛋白的表达来抑制系膜细胞增殖,影响细胞周期,有望为系膜增生性肾炎提供新的治疗靶点。
Objective:
To research the in vitro effects of Septin2 on Mesangial Proliferation in Rats.
Methods:
Cultured primary rat mesangial cells, processing the following experiment:1)The plasmids pEGFP-C 1 which could express the green fluorescent protein (GFP)were transfected into the cells. The transfection efficiency were determined by observing under fluorescent microscope 24 hours subsequently.2)Transfection of Septin2 plasmids by electroporation: Groups dividing:①pRK5 transfection group;②pRK5-Septin2 group. The plasmids pRK5 and pRK5-Setin2 were transfected into the mesangial cell under the optimal transfection conditions. The expression of Septin2 protein were examined by Western blot, cell cycle profiles were determined by flow cytometric analysis, the expressions of cell cycle regulatory proteins (cyclin D1、cyclin E、p21) were examined by Western blot.3) Design and synthesis the siRNA of GAPDH, according to different concentrations cells were divided into 4 groups:①siCon group,②200 nmol/L group,③400 nmol/L,④800 nmol/L group.48h later, Western blot to detect the expression of GAPDH protein.4) Electransfection of siSeptin2:Groups dividing:①siCon group,②siSeptin2 group. Western-blot detects the expression of Septin2 protein, flow cytometric analysis checks the cell cycle profiles.
Results:
1)When plasmids is 80μg, voltage is 340V, capacitor is 550μF, the electroporation transfection efficiency in primary rat mesangial cells can reach more than 50%.2)Compared to pRK5 transfection group, plasmids pRK5-Septin2 increased protein expression level of Septin2(P<0.05). Cell cycle analysis showed that the percentage of the cells in S stage were markedly reduced in pRK5-Septin2 group (P<0.05), the percentage of the cells in G0/G1 stage were markedly increased (P<0.05). The expression of cell cycle regulatory protein cyclin D1, cyclinE decreased and P21 reduced(P<0.05).3)When the concentration of siGAPDH was 800nmol/L, the inhibitory effect of protein level was the most significant (P<0.05).4) The synthesized siRNA of Septin2 gene inhibited the Septin2 protein expression levels (P<0.05), flow cytometry analysis didn't show any difference of significance.
Conclusion:
Overexpression of Septin2 is able to arrest cell cycle progression in the G0/G1 phase,the expression of positive regulators driving the G0/G1 phase, for example cyclin D1,cyclin E, were downregulated in cells transfected with Septin2, whereas expression of negative regulators p21 were upregulated. This is suggested that Septin2 may inhibite proliferation of mesangial cell by regulating the expression of cell cycle proteins. This may have provided a new target of mesangial proliferative glomerulonephritis therapy.
方法：培养原代大鼠肾小球系膜细胞,进行以下实验：1)将绿色荧光蛋白表达质粒pEGFP-C1用电穿孔转染的方法导入细胞,24h后荧光显微镜下观察绿色荧光蛋白表达情况。2)质粒电穿孔转染：细胞分组：①pRK5空质粒转染组,②pRK5-Septin2质粒转染组,分别进行电穿孔转染,Western blot检测两组细胞Septin2蛋白质表达水平,流式细胞仪分析Septin2对细胞周期进程的影响,Western blot检测系膜细胞周期调节蛋白(cyclin D1、cyclinE、p21)的表达变化。3)设计合成GAPDH的siRNA,依不同浓度分为①200nmol／L,②400nmol／L,③800nmol／L组,④无关对照转染组(siCon),电穿孔转染后培养48h,Western blot检测GAPDH蛋白质表达变化。4)siRNA电穿孔转染：细胞分组：①无关对照转染组(Sicon);②siSeptin2转染组。收集细胞蛋白,Western blot检测两组细胞Septin2蛋白质表达水平,流式细胞仪分析siSeptin2对细胞周期进程的影响。
结果：1)在80μg质粒,340V电压、550μF电容的电穿孔条件下,原代大鼠肾小球系膜细胞的电转染效率可以达到50%以上；2)与pRK5空质粒转染组相比,pRK5-Septin2质粒可使系膜细胞Septin2的蛋白质表达水平明显提高(P<0.05),流式细胞仪检测发现pRK5-Septin2质粒使G0／G1期细胞百分比明显增多而S期细胞百分比显著减少(P<0.05),同时细胞周期调节蛋白cyclin D1、cyclinE表达水平下调,p21的表达水平上调(P<0.05)；3) siGAPDH浓度为800nmol／L时蛋白质水平抑制效果最为显著(P<0.05)；4)与无关对照转染组(siCon)比较,siSeptin2转染组Septin2蛋白质表达水平明显降低,流式细胞仪检测结果显示两组细胞G0／G1期及S期细胞百分比无明显差异。
结论：Septin2过表达可使大鼠肾小球系膜细胞由G0／G1期向S期转化的百分比减少,细胞周期正调控蛋白表达减少而负调控蛋白表达增多,提示可能通过调节细胞周期蛋白的表达来抑制系膜细胞增殖,影响细胞周期,有望为系膜增生性肾炎提供新的治疗靶点。
Objective:
To research the in vitro effects of Septin2 on Mesangial Proliferation in Rats.
Methods:
Cultured primary rat mesangial cells, processing the following experiment:1)The plasmids pEGFP-C 1 which could express the green fluorescent protein (GFP)were transfected into the cells. The transfection efficiency were determined by observing under fluorescent microscope 24 hours subsequently.2)Transfection of Septin2 plasmids by electroporation: Groups dividing:①pRK5 transfection group;②pRK5-Septin2 group. The plasmids pRK5 and pRK5-Setin2 were transfected into the mesangial cell under the optimal transfection conditions. The expression of Septin2 protein were examined by Western blot, cell cycle profiles were determined by flow cytometric analysis, the expressions of cell cycle regulatory proteins (cyclin D1、cyclin E、p21) were examined by Western blot.3) Design and synthesis the siRNA of GAPDH, according to different concentrations cells were divided into 4 groups:①siCon group,②200 nmol/L group,③400 nmol/L,④800 nmol/L group.48h later, Western blot to detect the expression of GAPDH protein.4) Electransfection of siSeptin2:Groups dividing:①siCon group,②siSeptin2 group. Western-blot detects the expression of Septin2 protein, flow cytometric analysis checks the cell cycle profiles.
Results:
1)When plasmids is 80μg, voltage is 340V, capacitor is 550μF, the electroporation transfection efficiency in primary rat mesangial cells can reach more than 50%.2)Compared to pRK5 transfection group, plasmids pRK5-Septin2 increased protein expression level of Septin2(P<0.05). Cell cycle analysis showed that the percentage of the cells in S stage were markedly reduced in pRK5-Septin2 group (P<0.05), the percentage of the cells in G0/G1 stage were markedly increased (P<0.05). The expression of cell cycle regulatory protein cyclin D1, cyclinE decreased and P21 reduced(P<0.05).3)When the concentration of siGAPDH was 800nmol/L, the inhibitory effect of protein level was the most significant (P<0.05).4) The synthesized siRNA of Septin2 gene inhibited the Septin2 protein expression levels (P<0.05), flow cytometry analysis didn't show any difference of significance.
Conclusion:
Overexpression of Septin2 is able to arrest cell cycle progression in the G0/G1 phase,the expression of positive regulators driving the G0/G1 phase, for example cyclin D1,cyclin E, were downregulated in cells transfected with Septin2, whereas expression of negative regulators p21 were upregulated. This is suggested that Septin2 may inhibite proliferation of mesangial cell by regulating the expression of cell cycle proteins. This may have provided a new target of mesangial proliferative glomerulonephritis therapy.
引文
[1]黎磊石,关天俊,刘志红,等。4298例成年人肾小球疾病病理类型及流行病学特点[J].肾脏病与透析肾移植杂志,1997,6(2)：103～109.
[2]马玉凤,孝文泉,赵利。195例肾小球疾病的临床病理分型与中医辨证分型的相关分析[J].北京中医药大学学报,1998,5(21)：48～50.
[3]Cove-Smith, Hendry BM.The regulation of mesangial cell proliferation. Nephron Exp Nephrol,2008,108:74-79.
[4]Qiu LQ, Sinniah R, Hsu SI. Role of differential and cell type-specific expression of cell cycle regulatory proteins in mediating progressive glomerular injury in human IgA nephropathy. Lab Invest,2004,84 (9):1112～1125.
[5]Floege J, Eitner F, Alpers CE. A new look at platelet-derived growth factor in renal disease. J Am Soc Nephrol,2008,19 (1):12～23.
[6]Ghosh PM, Mikhailova M, Bedolla R, et al. Arginine vasopressin stimulates mesangial cell proliferation by activating the epidermal growth factor receptor. Am J Physiol Renal Physiol, 2001,280 (6):972～979.
[7]Breck Byers, Loretta Goetsch. A highly ordered ring of membrane-associated filaments in budding yeast. Cell Biology,1976,69:717～721.
[8]Hall PA,Jung K,Hillan KJ,et al. Expression profiling the human septin gene family.J Pathol.2005,206:269～271.
[9]Martinez C,Sanjuan MA,Dent JA,et al. Human septin-septin interaction as a prerequisite for targeting septin complex in the cytosol. Biochem J,2004,382:783～791.
[10]Kinoshita M, Kumar S, Mizoguchi A, Ide C, Kinoshita A, Haraguchi T, et al. Nedd5, a mammalian septin, is a novel cytoskeletal component interacting with actin-based structures. Genes Dev.1997,11:1535-47.
[11]Preeti Ahuja, Evelyne Perriard, William Trimbleb, et al. Probing the role of septins in cardiomyocytes. Experimental Cell Research 2006:1598～1609.
[12]Jin-Liang Zhu,Sheng-Li Lin,Mo Li et al.Septin2 is modifed by SUMOylation and required for chromosome congression in mouse oocytes. Cell Cycle 2010,9:1-10.
[13]张军,王胜建,唐小柯等。septin2基因短剪接本转表达产物对小鼠皮肤培养细胞作用的实验研究,中华整形外科杂志,2007,23：147～150.
[14]Jaroszeski MJ, Dang V, Pottinger C, et al. Toxicity of anticancer agents media ted by electroporation in vitrol. Anticancer Drugs,2000.11:201～208
[15]Mene P.Mesangial cell cultures.[J]. Nephrol,2001,14:198～203.
[16]Gene Gulser Electroporation System Instruction Mannual Bio—RAD
[17]Yang R, Bie W, Haegebarth A, Tyner AL. Differential regulation of D-type cyclins in the mouse intestine. Cell Cycle 2006; 5:180～183.
[18]Zhi-fan Jia, Qiang Huang, Chun-sheng Kang, et al. Overexpression of septin 7 suppresses glioma cell growth. J Neurooncol 2009.
[19]Brandon E. Kremer, Laura A. Adang, Ian G. Macara. Septins Regulate Actin Organization and Cell Cycle Arrest Through SOCS7-Mediated Nuclear Accumulation of NCK. Cell. 2007,130(5):837～850.
[20]Sirajuddin M, Farkasovsky M, Hauer F, et al. Structural insight into filament formation by mammalian septins. Nature,2007,449:311～315.
[1]Adam JC, Pringle JR, Peifer M. Evidence for functional differentiation among Drosophila septins in cytokinesis and cellularization. Mol Biol Cell,2000,11:3123～3135.
[2]Barral Y, Mermall V, Mooseker MS, et al. Compartmentalization of the cell cortex by septins is required for maintenance of cell polarity in yeast. Mol Cell,2000,5:841～851.
[3]Beites CL, Xie H, Bowser R, et al. The septin CDCrel-1 binds syntaxin and inhibits exocytosis. Nat Neurosci,1999,2:434～439.
[4]Bourne HR, Sanders DA, McCormick F. The GTPase superfamily:conserved structure and molecular mechanism. Nature,1991,349:117～127.
[5]Hall PA,Jung K,Hillan KJ,et al. Expression profiling the human septin gene family.J Pathol,2005,206:269～278.
[6]Peter A Hall, SE Hilary Russell. The pathobiology of the septin gene family. Journal of Pathology J Pathol,2004,204:489～505.
[7]Martinez C,Sanjuan MA,Dent JA,et al. Human septin-septin interaction as a prerequisite for targeting septin complex in the cytosol. Biochem J,2004,382:783～791.
[8]Saraste M, Sibbald PR, Wittinghofer A. The P-loop—a common motif in ATP-and GTP-binding proteins. Trends Biochem Sci,1990,15:430～434.
[9]Sirajuddin M, Farkasovsky M, Hauer F, et al. Structural insight into filament formation by mammalian septins. Nature,2007,449:311～315.
[10]Kinoshita M. Assembly of mammalian septins. J Biochem,2003,134:491～496.
[11]Hartwell LH. Genetic control of the cell division cycle in yeast (Ⅳ). Genes controlling bud emergence and cytokinesis. Exp Cell Res,1971,69:265～276.
[12]Gladfelter AS, Pringle JR, Lew DJ. The septin cortex at the yeast mother-bud neck. Curr Opin Microbiol,2001,4:681～685.
[13]Kusch J, Meyer A, Snyder MP, Barral Y. Microtubule capture by the cleavage apparatus is required for proper spindle positioning in yeast. Genes Dev,2002,16: 1627～1639.
[14]Longtine MS, Bi E. Regulation of septin organization and function in yeast. Trends Cell Biol,2003,13:403～410.
[15]Sheffield PJ,Oliver CJ,Kremer BE,et al. Borg/septin interactions and the assembly of Mammalian septin heterodimers,trimers,and filaments. J Biol Chem,2003,278:348～353.
[16]Kremer BE,Haystead T,Macara IG. Mammalian septin regulate microtubule stability though binding protein MAP4. Mol Biol Cell,2005,16:4648～4659.
[17]Spiliotis, Kinoshita M, Nelson WJ. A mitotic septin scaffold required for Mammalian chromosome congression and segregation. Science,2005,307:1781～1785.
[18]Kinoshita M,Field CM,Coughlin ML,et al. Self-and actin-templated assembly of Mammalian septins. Dev Cell,2002,3:791～802.
[19]Edgar R. Gomes,Shantanu Jani,Gregg G. Gundersen.MSF-A interacts with hypoxia-inducible factor-lalpha and augments hypoxia-inducible factor transcriptional activation to affect tumorigenicity and angiogenesis. Cell,2005,121:451～463.
[20]Julien Caul,Alan Hall.Cdc42 controls the polarity of the actin and microtubule cytoskeletons through two distinct signal transduction pathways. Cell Science,118:2579～2587.
[21]Chant J. septin scaffolds and cleavage planes in Saccharomyces. Cell,1996,84: 187～190.
[22]Dong-Seok Kim, Sherri-Lynn Hubbard,Aurelia Peraud,et al.Analysis of Mammalian Septin Expression in HumanMalignant Brain Tumors. Canada Neoplasia.2004, 6(2):168～178.
[23]Preeti Ahujaa, Evelyne erriarda, et al. Probing the role of septins in cardiomyocytes Experimental cell research,2006,1598～1609.
[24]Kinoshita M, Kumar S, Mizoguchi A, et al. Nedd5, a mammalian septin, is a novel cytoskeletal component interacting with actin-based structures.Genes Dev,1997,11: 1535～1547.
[25]Surka MC, Tsang CW, Trimble WS. The mammalian septin MSF localizes with microtubules and is required for completion of cytokinesis. Mol Biol Cell,2002,13: 3532-3545.
[26]Robertson C, Church SW, Nagar HA, et al. Properties of SEPT9 isoforms and the requirement for GTP binding. J Pathol 2004,203:519-527.
[27]Zhi-fan Jia, Qiang Huang, Chun-sheng Kang,et al. Overexpression of septin 7 suppresses glioma cell growth.. J Neurooncol.
[28]Maria E. Gonzalezl, Olga Makaroval, Esther A,et al. Up-regulation of SEPT9_vl stabilizes c-Jun-N-Terminal Kinase and contributes to its pro-proliferative activity in mammary epithelial cells. Cell Signal,2009,21:477-487.
[29]Sharon Amir,Ruoxiang Wang,Haim Matzkin, et al. MSF-A Interacts with Hypoxia-Inducible Factor-1A and Augments Hypoxia-Inducible Factor Transcriptional Activation to Affect Tumorigenicity and Angiogenesis. Cancer Res,2006,66:856-866.
[30]Katrina M. Comerford, Eoin P. Cummins,et al. c-Jun NH2-Terminal Kinase Activation Contributes to Hypoxia-Inducible Factor la-Dependent P-Glycoprotein Expression in Hypoxia. Cancer Research,2004,64:9057-9061.
[31]Ron Wisdom, Randall S.Johnson, Connie Moore. c-Jun regulates cell cycle progression and apoptosis by distinct mechanisms. The EMBO Journal 1999,18:188-197.
[32]Zhang EY, DeTure MA, Bubb MR, Caviston TL, Self-assembly of the brain MAP-2 microtubule-binding region into polymeric structures resembling Alzheimer filaments. Biochem Biophys Res Commun.1996,229:176-181.
[33]Whitfield ML, Sherlock G, Saldanha AJ, et al. Identification of genes periodically expressed in the human cell cycle and theirexpression in tumors. Mol Biol Cell 2002,13: 1977-2000.
[34]Kinoshita M, Field CM, Coughlin ML, et al. Self-and actin-templated assembly of mammalian septins. Dev Cell 2002; 3:791-802.
[35]Schmidt, Nichols. A barrier to lateral diffusion in the cleavage furrow of dividing mammalian cells. Curr Biol. 2004,14:1002-1006.
[36]Kartmann B, RothD.Novel roles for mammalian septins:from vesicle trafficking to oncogenesis. J Cell Sci,2001,114:839-844.
[37]Elias T. Spiliotis, W. James Nelson.Here come the septins:novel polymers that coordinate intracellular functions and organization Journal of Cell Science 2005,119:4-10.
[38]Kinoshita A, Kinoshita M, Akiyama H, et al. Identification of septins in neurofibrillary tangles in Alzheimer's disease. Am J Pathol 1998; 153:1551-1560.
[39]Cheon MS, Fountoulakis M, Dierssen M, et al.Expression profiles of proteins in fetal brain with Down syndrome. J Neural Transm Suppl 2001; 61:311-319.
[40]Sakai K, Kurimoto M, Tsugu A, Hubbard SL, Trimble WS, Rutka JT. Expression of Nedd5, a mammalian septin, in human brain tumors. J Neurooncol 2002; 57:169～177.
[41]Kalikin LM, Sims HL, Petty EM. Genomic and expression analyses of alternatively spliced transcripts of the MLL septin-like fusion gene (MSF) that map to a 17q25 region of loss in breast and ovarian tumors. Genomics 2000; 63:165～172.
[42]Russell SE, McIlhatton MA, Burrows JF, et al. Isolation and mapping of a human septin gene to a region on chromosome 17q, commonly deleted in sporadic epithelial ovarian tumors. Cancer Res,2000; 60:4729～4734.
[43]Proteomic identification of a role for the von Hippel Lindau tumour suppressor in changes in the expression of mitochondrial proteins and septin2 in renal cell carcinoma. Proteomics,2006,6:3880～3893.
[44]Kim DS, Hubbard SL, Peraud A, Salhia B, Sakai K, Rutka, JT. Analysis of mammalian septin expression in human malignant brain tumors. Neoplasia,2004,6:168～178.
[45]Liu M, Shen S, Chen F, et al. Linking the septin expression with carcinogenesis, Mol Biol Rep,2010 [Epub ahead of print].
[2]马玉凤,孝文泉,赵利。195例肾小球疾病的临床病理分型与中医辨证分型的相关分析[J].北京中医药大学学报,1998,5(21)：48～50.
[3]Cove-Smith, Hendry BM.The regulation of mesangial cell proliferation. Nephron Exp Nephrol,2008,108:74-79.
[4]Qiu LQ, Sinniah R, Hsu SI. Role of differential and cell type-specific expression of cell cycle regulatory proteins in mediating progressive glomerular injury in human IgA nephropathy. Lab Invest,2004,84 (9):1112～1125.
[5]Floege J, Eitner F, Alpers CE. A new look at platelet-derived growth factor in renal disease. J Am Soc Nephrol,2008,19 (1):12～23.
[6]Ghosh PM, Mikhailova M, Bedolla R, et al. Arginine vasopressin stimulates mesangial cell proliferation by activating the epidermal growth factor receptor. Am J Physiol Renal Physiol, 2001,280 (6):972～979.
[7]Breck Byers, Loretta Goetsch. A highly ordered ring of membrane-associated filaments in budding yeast. Cell Biology,1976,69:717～721.
[8]Hall PA,Jung K,Hillan KJ,et al. Expression profiling the human septin gene family.J Pathol.2005,206:269～271.
[9]Martinez C,Sanjuan MA,Dent JA,et al. Human septin-septin interaction as a prerequisite for targeting septin complex in the cytosol. Biochem J,2004,382:783～791.
[10]Kinoshita M, Kumar S, Mizoguchi A, Ide C, Kinoshita A, Haraguchi T, et al. Nedd5, a mammalian septin, is a novel cytoskeletal component interacting with actin-based structures. Genes Dev.1997,11:1535-47.
[11]Preeti Ahuja, Evelyne Perriard, William Trimbleb, et al. Probing the role of septins in cardiomyocytes. Experimental Cell Research 2006:1598～1609.
[12]Jin-Liang Zhu,Sheng-Li Lin,Mo Li et al.Septin2 is modifed by SUMOylation and required for chromosome congression in mouse oocytes. Cell Cycle 2010,9:1-10.
[13]张军,王胜建,唐小柯等。septin2基因短剪接本转表达产物对小鼠皮肤培养细胞作用的实验研究,中华整形外科杂志,2007,23：147～150.
[14]Jaroszeski MJ, Dang V, Pottinger C, et al. Toxicity of anticancer agents media ted by electroporation in vitrol. Anticancer Drugs,2000.11:201～208
[15]Mene P.Mesangial cell cultures.[J]. Nephrol,2001,14:198～203.
[16]Gene Gulser Electroporation System Instruction Mannual Bio—RAD
[17]Yang R, Bie W, Haegebarth A, Tyner AL. Differential regulation of D-type cyclins in the mouse intestine. Cell Cycle 2006; 5:180～183.
[18]Zhi-fan Jia, Qiang Huang, Chun-sheng Kang, et al. Overexpression of septin 7 suppresses glioma cell growth. J Neurooncol 2009.
[19]Brandon E. Kremer, Laura A. Adang, Ian G. Macara. Septins Regulate Actin Organization and Cell Cycle Arrest Through SOCS7-Mediated Nuclear Accumulation of NCK. Cell. 2007,130(5):837～850.
[20]Sirajuddin M, Farkasovsky M, Hauer F, et al. Structural insight into filament formation by mammalian septins. Nature,2007,449:311～315.
[1]Adam JC, Pringle JR, Peifer M. Evidence for functional differentiation among Drosophila septins in cytokinesis and cellularization. Mol Biol Cell,2000,11:3123～3135.
[2]Barral Y, Mermall V, Mooseker MS, et al. Compartmentalization of the cell cortex by septins is required for maintenance of cell polarity in yeast. Mol Cell,2000,5:841～851.
[3]Beites CL, Xie H, Bowser R, et al. The septin CDCrel-1 binds syntaxin and inhibits exocytosis. Nat Neurosci,1999,2:434～439.
[4]Bourne HR, Sanders DA, McCormick F. The GTPase superfamily:conserved structure and molecular mechanism. Nature,1991,349:117～127.
[5]Hall PA,Jung K,Hillan KJ,et al. Expression profiling the human septin gene family.J Pathol,2005,206:269～278.
[6]Peter A Hall, SE Hilary Russell. The pathobiology of the septin gene family. Journal of Pathology J Pathol,2004,204:489～505.
[7]Martinez C,Sanjuan MA,Dent JA,et al. Human septin-septin interaction as a prerequisite for targeting septin complex in the cytosol. Biochem J,2004,382:783～791.
[8]Saraste M, Sibbald PR, Wittinghofer A. The P-loop—a common motif in ATP-and GTP-binding proteins. Trends Biochem Sci,1990,15:430～434.
[9]Sirajuddin M, Farkasovsky M, Hauer F, et al. Structural insight into filament formation by mammalian septins. Nature,2007,449:311～315.
[10]Kinoshita M. Assembly of mammalian septins. J Biochem,2003,134:491～496.
[11]Hartwell LH. Genetic control of the cell division cycle in yeast (Ⅳ). Genes controlling bud emergence and cytokinesis. Exp Cell Res,1971,69:265～276.
[12]Gladfelter AS, Pringle JR, Lew DJ. The septin cortex at the yeast mother-bud neck. Curr Opin Microbiol,2001,4:681～685.
[13]Kusch J, Meyer A, Snyder MP, Barral Y. Microtubule capture by the cleavage apparatus is required for proper spindle positioning in yeast. Genes Dev,2002,16: 1627～1639.
[14]Longtine MS, Bi E. Regulation of septin organization and function in yeast. Trends Cell Biol,2003,13:403～410.
[15]Sheffield PJ,Oliver CJ,Kremer BE,et al. Borg/septin interactions and the assembly of Mammalian septin heterodimers,trimers,and filaments. J Biol Chem,2003,278:348～353.
[16]Kremer BE,Haystead T,Macara IG. Mammalian septin regulate microtubule stability though binding protein MAP4. Mol Biol Cell,2005,16:4648～4659.
[17]Spiliotis, Kinoshita M, Nelson WJ. A mitotic septin scaffold required for Mammalian chromosome congression and segregation. Science,2005,307:1781～1785.
[18]Kinoshita M,Field CM,Coughlin ML,et al. Self-and actin-templated assembly of Mammalian septins. Dev Cell,2002,3:791～802.
[19]Edgar R. Gomes,Shantanu Jani,Gregg G. Gundersen.MSF-A interacts with hypoxia-inducible factor-lalpha and augments hypoxia-inducible factor transcriptional activation to affect tumorigenicity and angiogenesis. Cell,2005,121:451～463.
[20]Julien Caul,Alan Hall.Cdc42 controls the polarity of the actin and microtubule cytoskeletons through two distinct signal transduction pathways. Cell Science,118:2579～2587.
[21]Chant J. septin scaffolds and cleavage planes in Saccharomyces. Cell,1996,84: 187～190.
[22]Dong-Seok Kim, Sherri-Lynn Hubbard,Aurelia Peraud,et al.Analysis of Mammalian Septin Expression in HumanMalignant Brain Tumors. Canada Neoplasia.2004, 6(2):168～178.
[23]Preeti Ahujaa, Evelyne erriarda, et al. Probing the role of septins in cardiomyocytes Experimental cell research,2006,1598～1609.
[24]Kinoshita M, Kumar S, Mizoguchi A, et al. Nedd5, a mammalian septin, is a novel cytoskeletal component interacting with actin-based structures.Genes Dev,1997,11: 1535～1547.
[25]Surka MC, Tsang CW, Trimble WS. The mammalian septin MSF localizes with microtubules and is required for completion of cytokinesis. Mol Biol Cell,2002,13: 3532-3545.
[26]Robertson C, Church SW, Nagar HA, et al. Properties of SEPT9 isoforms and the requirement for GTP binding. J Pathol 2004,203:519-527.
[27]Zhi-fan Jia, Qiang Huang, Chun-sheng Kang,et al. Overexpression of septin 7 suppresses glioma cell growth.. J Neurooncol.
[28]Maria E. Gonzalezl, Olga Makaroval, Esther A,et al. Up-regulation of SEPT9_vl stabilizes c-Jun-N-Terminal Kinase and contributes to its pro-proliferative activity in mammary epithelial cells. Cell Signal,2009,21:477-487.
[29]Sharon Amir,Ruoxiang Wang,Haim Matzkin, et al. MSF-A Interacts with Hypoxia-Inducible Factor-1A and Augments Hypoxia-Inducible Factor Transcriptional Activation to Affect Tumorigenicity and Angiogenesis. Cancer Res,2006,66:856-866.
[30]Katrina M. Comerford, Eoin P. Cummins,et al. c-Jun NH2-Terminal Kinase Activation Contributes to Hypoxia-Inducible Factor la-Dependent P-Glycoprotein Expression in Hypoxia. Cancer Research,2004,64:9057-9061.
[31]Ron Wisdom, Randall S.Johnson, Connie Moore. c-Jun regulates cell cycle progression and apoptosis by distinct mechanisms. The EMBO Journal 1999,18:188-197.
[32]Zhang EY, DeTure MA, Bubb MR, Caviston TL, Self-assembly of the brain MAP-2 microtubule-binding region into polymeric structures resembling Alzheimer filaments. Biochem Biophys Res Commun.1996,229:176-181.
[33]Whitfield ML, Sherlock G, Saldanha AJ, et al. Identification of genes periodically expressed in the human cell cycle and theirexpression in tumors. Mol Biol Cell 2002,13: 1977-2000.
[34]Kinoshita M, Field CM, Coughlin ML, et al. Self-and actin-templated assembly of mammalian septins. Dev Cell 2002; 3:791-802.
[35]Schmidt, Nichols. A barrier to lateral diffusion in the cleavage furrow of dividing mammalian cells. Curr Biol. 2004,14:1002-1006.
[36]Kartmann B, RothD.Novel roles for mammalian septins:from vesicle trafficking to oncogenesis. J Cell Sci,2001,114:839-844.
[37]Elias T. Spiliotis, W. James Nelson.Here come the septins:novel polymers that coordinate intracellular functions and organization Journal of Cell Science 2005,119:4-10.
[38]Kinoshita A, Kinoshita M, Akiyama H, et al. Identification of septins in neurofibrillary tangles in Alzheimer's disease. Am J Pathol 1998; 153:1551-1560.
[39]Cheon MS, Fountoulakis M, Dierssen M, et al.Expression profiles of proteins in fetal brain with Down syndrome. J Neural Transm Suppl 2001; 61:311-319.
[40]Sakai K, Kurimoto M, Tsugu A, Hubbard SL, Trimble WS, Rutka JT. Expression of Nedd5, a mammalian septin, in human brain tumors. J Neurooncol 2002; 57:169～177.
[41]Kalikin LM, Sims HL, Petty EM. Genomic and expression analyses of alternatively spliced transcripts of the MLL septin-like fusion gene (MSF) that map to a 17q25 region of loss in breast and ovarian tumors. Genomics 2000; 63:165～172.
[42]Russell SE, McIlhatton MA, Burrows JF, et al. Isolation and mapping of a human septin gene to a region on chromosome 17q, commonly deleted in sporadic epithelial ovarian tumors. Cancer Res,2000; 60:4729～4734.
[43]Proteomic identification of a role for the von Hippel Lindau tumour suppressor in changes in the expression of mitochondrial proteins and septin2 in renal cell carcinoma. Proteomics,2006,6:3880～3893.
[44]Kim DS, Hubbard SL, Peraud A, Salhia B, Sakai K, Rutka, JT. Analysis of mammalian septin expression in human malignant brain tumors. Neoplasia,2004,6:168～178.
[45]Liu M, Shen S, Chen F, et al. Linking the septin expression with carcinogenesis, Mol Biol Rep,2010 [Epub ahead of print].