Mel-18在子宫内膜癌中的表达及临床意义
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摘要
目的:探讨黑色素瘤蛋白(Mel-18)在子宫内膜癌(EC)中的表达及其在肿瘤细胞恶性生物学行为中的作用及机制。方法:免疫组化染色法(IHC)和实时荧光定量聚合酶链式反应(RT-qPCR)检测Mel-18在EC组织及正常子宫内膜组织中的表达。小干扰(siRNA)及腺病毒下调或上调Mel-18在EC细胞系中的表达,MTT法检测Mel-18对EC细胞增殖能力的影响;Transwell小室实验检测Mel-18对EC细胞迁移能力的影响;Western blot法检测相关目的蛋白的表达。结果:IHC和RT-qPCR结果显示,Mel-18在EC组织中显著高表达(P<0.05);腺病毒上调Mel-18在EC细胞系中的表达后,细胞的增殖、迁移能力升高,PI3K/Akt/mTOR信号转导通路表达升高;siRNA下调Mel-18表达抑制细胞的增殖、迁移能力,PI3K/Akt/mTOR信号转导通路表达降低。结论:Mel-18在EC中高表达,在EC的发生发展中可能起到促癌基因的作用,可通过活化PI3K/Akt/mTOR信号转导通路促进EC细胞增殖、迁移能力。
Objective: To detect the expression of Melanoma nuclear protein 18( Mel-18) in endometrial cancer( EC) and evaluate the biological effects of Mel-18 on the proliferation and immigration of EC. Methods: Immunohistochemistry( IHC) and reverse transcription c quantitative polymerase chain reaction( RT-qPCR) assays were used to examine the expression of Mel-18 in EC. Adenovirus and small interfering RNA( siRNA) were used to regulate Mel-18 gene levels in cells.MTT were used to detect the cell proliferation ability.Transwell migration assay was used to detect the cell immigration ability.Western blot was used to detect the related proteins expression. Results: Mel-18 mRNA and protein were both highly expressed in EC( P<0.05).Up-regulation of Mel-18 was significantly promoted the cell proliferation and migration capacity.Loss of Mel-18 was inhibited the cell proliferation and migration ability.We also explored the potential mechanism of Mel-18 in EC cell lines.Overexpression of Mel-18 activated the PI3K/Akt/mT OR pathway,the expressions of PI3K p85α,p-Akt,p-mTOR proteins were significantly increased.Loss of Mel-18 inactivated the PI3K/Akt/mT OR pathway. Conclusion:Mel-18 was highly expressed in EC and promoted cell proliferation and migration via PI3K/Akt/mT OR pathway.
Objective: To detect the expression of Melanoma nuclear protein 18( Mel-18) in endometrial cancer( EC) and evaluate the biological effects of Mel-18 on the proliferation and immigration of EC. Methods: Immunohistochemistry( IHC) and reverse transcription c quantitative polymerase chain reaction( RT-qPCR) assays were used to examine the expression of Mel-18 in EC. Adenovirus and small interfering RNA( siRNA) were used to regulate Mel-18 gene levels in cells.MTT were used to detect the cell proliferation ability.Transwell migration assay was used to detect the cell immigration ability.Western blot was used to detect the related proteins expression. Results: Mel-18 mRNA and protein were both highly expressed in EC( P<0.05).Up-regulation of Mel-18 was significantly promoted the cell proliferation and migration capacity.Loss of Mel-18 was inhibited the cell proliferation and migration ability.We also explored the potential mechanism of Mel-18 in EC cell lines.Overexpression of Mel-18 activated the PI3K/Akt/mT OR pathway,the expressions of PI3K p85α,p-Akt,p-mTOR proteins were significantly increased.Loss of Mel-18 inactivated the PI3K/Akt/mT OR pathway. Conclusion:Mel-18 was highly expressed in EC and promoted cell proliferation and migration via PI3K/Akt/mT OR pathway.
引文
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[14] Wiederschain D,Chen L,Johnson B,et al.Contribution of polycomb homologues Bmi-1 and Mel-18 to medulloblastoma pathogenesis[J].Mol Cell Biol,2007,27(13):4968-4979
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[18] Honig A,Weidler C,Hausler S,et al.Overexpression of polycomb protein BMI-1 in human specimens of breast,ovarian,endometrial and cervical cancer[J].Anticancer Res,2010,30(5):1559-1564
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[20] Ji H,Cao M,Ren K,Et al.Expression and clinicopathological significance of Mel-18 and Bmi-1 in esophageal squamous cell carcinoma[J].Technol Cancer Res Treat,2017:1533034617705055
[21] Tagawa M,Sakamoto T,Shigemoto K,et al.Expression of novel DNA-binding protein with zinc finger structure in various tumor-cells[J].J Biol Chem,1990,265(32):20021-20026
[22] Guo WJ,Zeng MS,Yadav A,et al.Mel-18 acts as a tumor suppressor by repressing Bmi-1 expression and down-regulating Akt activity in breast cancer cells[J].Cancer Research,2007,67(11):5083-5089
[23] Guo WJ,Datta S,Band V,et al.Mel-18,a polycomb group protein,regulates cell proliferation and senescence via transcriptional repression of Bmi-1 and c-Myc oncoproteins[J].Mol Biol Cell,2007,18(2):536-546
[24] Lee JY,Jang KS,Shin DH,et al.Mel-18 negatively regulates INK4a/ARF-independent cell cycle progression via Akt inactivation in breast cancer[J].Cancer Res,2008,68(11):4201-4209
[25] Chun T,Rho SB,Byun HJ,et al.The polycomb group gene product Mel-18 interacts with cyclin D2 and modulates its activity[J].FEBS Lett,2005,579(24):5275-5280
[26] Mabuchi S,Kuroda H,Takahashi R,et al.The PI3K/AKT/mTOR pathway as a therapeutic target in ovarian cancer[J].Gynecol Oncol,2015,137(1):173-17
[27] Jung JH,Choi HJ,Maeng YS,et al.Mel-18,a mammalian Polycomb gene,regulates angiogenic gene expression of endothelial cells[J].Biochem Biophys Res Commun,2010,400(4):523-530
[28] 刘黎琼,刘泽林,崔海燕,等.抑制mel18表达增强桂皮醛诱导的HL60细胞分化[J].中国病理生理杂志,2017,33(10):1852-1857
[2] 王遥.子宫内膜癌分子分型及临床应用研究进展[J].现代妇产科进展,2018,27(7):549-551
[3] Zhou M,Zhang Z,Zhao H,et al.A novel lncRNA-focus expression signature for survival prediction in endometrial carcinoma[J].BMC Cancer,2018,18(1):39
[4] Dukers DF,van Galen JC,Giroth C,et al.Unique polycomb gene expression pattern in hodgkin's lymphoma and hodgkin's lymphoma-derived cell lines[J].Am J Pathol,2004,164(3):873-881
[5] Lessard J,Sauvageau G.Polycomb group genes as epigenetic regulators of normal and leukemic hemopoiesis[J].Exp Hematol,2003,31(7):567-585
[6] Abd al kader L,oka T,Takata K,et al.In aggressive variants of non-Hodgkin lymphomas,Ezh2 is strongly expressed and polycomb repressive complex PRC1.4 dominates over PRC1.2[J].Virchows Arch,2013,463(5):697-711
[7] Riis ML,Luders T,Nesbakken AJ,et al.Expression of BMI-1 and Mel-18 in breast tissue--a diagnostic marker in patients with breast cancer[J].BMC Cancer,2010,10:686
[8] Lee JY,Park MK,Park JH,et al.Loss of the polycomb protein Mel-18 enhances the epithelial-mesenchymal transition by ZEB1 and ZEB2 expression through the downregulation of miR-205 in breast cancer[J].Oncogene,2014,33(10):1325-1335
[9] Zhang X W,Sheng Y P,Li Q,et al.BMI1 and Mel-18 oppositely regulate carcinogenesis and progression of gastric cancer[J].Mol Cancer,2010,9:40
[10] Wang XF,Zhang XW,Hua RX,et al.Mel-18 negatively regulates stem cell-like properties through downregulation of miR-21 in gastric cancer[J].Oncotarget,2016,7(39):63352-63361
[11] Wang W,Lin T,Huang J,et al.Analysis of Mel-18 expression in prostate cancer tissues and correlation with clinicopathologic features[J].Urol Oncol,2011,29(3):244-251
[12] Wang W,Yuasa T,Tsuchiya N,et al.The novel tumor-suppressor Mel-18 in prostate cancer:its functional polymorphism,expression and clinical significance[J].Int J Cancer,2009,125(12):2836-2843
[13] Dukers DF,Van galen JC,Giroth C,et al.Unique Polycomb Gene Expression Pattern in Hodgkin's Lymphoma and Hodgkin's Lymphoma-Derived Cell Lines[J].Am J Pathol,2004,164(3):873-881
[14] Wiederschain D,Chen L,Johnson B,et al.Contribution of polycomb homologues Bmi-1 and Mel-18 to medulloblastoma pathogenesis[J].Mol Cell Biol,2007,27(13):4968-4979
[15] Park JH,Lee JY,Shin DH,et al.Loss of Mel-18 induces tumor angiogenesis through enhancing the activity and expression of HIF-1alpha mediated by the PTEN/PI3K/Akt pathway[J].Oncogene,2011,30(45):4578-4589
[16] Kanno M,Hasegawa M,Ishida A,et al.mel-18,a Polycomb group-related mammalian gene,encodes a transcriptional negative regulator with tumor suppressive activity[J].EMBO J,1995,14(22):5672-5678
[17] Violante C,Chiodo L,Conte A M,et al.Si(111)2×1 surface isomers:DFT investigations on stability and doping effects[J].Surface Science,2014,621(4):123-127
[18] Honig A,Weidler C,Hausler S,et al.Overexpression of polycomb protein BMI-1 in human specimens of breast,ovarian,endometrial and cervical cancer[J].Anticancer Res,2010,30(5):1559-1564
[19] Goebl MG.The bmi1 and me18 gene products define a new family of dna binding proteins involved in cell proliferation and tumorigenesis[J].Cell,1991,66(4):623
[20] Ji H,Cao M,Ren K,Et al.Expression and clinicopathological significance of Mel-18 and Bmi-1 in esophageal squamous cell carcinoma[J].Technol Cancer Res Treat,2017:1533034617705055
[21] Tagawa M,Sakamoto T,Shigemoto K,et al.Expression of novel DNA-binding protein with zinc finger structure in various tumor-cells[J].J Biol Chem,1990,265(32):20021-20026
[22] Guo WJ,Zeng MS,Yadav A,et al.Mel-18 acts as a tumor suppressor by repressing Bmi-1 expression and down-regulating Akt activity in breast cancer cells[J].Cancer Research,2007,67(11):5083-5089
[23] Guo WJ,Datta S,Band V,et al.Mel-18,a polycomb group protein,regulates cell proliferation and senescence via transcriptional repression of Bmi-1 and c-Myc oncoproteins[J].Mol Biol Cell,2007,18(2):536-546
[24] Lee JY,Jang KS,Shin DH,et al.Mel-18 negatively regulates INK4a/ARF-independent cell cycle progression via Akt inactivation in breast cancer[J].Cancer Res,2008,68(11):4201-4209
[25] Chun T,Rho SB,Byun HJ,et al.The polycomb group gene product Mel-18 interacts with cyclin D2 and modulates its activity[J].FEBS Lett,2005,579(24):5275-5280
[26] Mabuchi S,Kuroda H,Takahashi R,et al.The PI3K/AKT/mTOR pathway as a therapeutic target in ovarian cancer[J].Gynecol Oncol,2015,137(1):173-17
[27] Jung JH,Choi HJ,Maeng YS,et al.Mel-18,a mammalian Polycomb gene,regulates angiogenic gene expression of endothelial cells[J].Biochem Biophys Res Commun,2010,400(4):523-530
[28] 刘黎琼,刘泽林,崔海燕,等.抑制mel18表达增强桂皮醛诱导的HL60细胞分化[J].中国病理生理杂志,2017,33(10):1852-1857