上调PHLPP1表达对食管癌细胞增殖、黏附、侵袭和迁移能力的影响
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摘要
目的:研究上调PHLPP1表达对食管癌细胞增殖、黏附、侵袭和迁移能力的影响。方法:用qRT-PCR和Western blot法检测食管癌细胞Eca109、EC9706、KYSE510和正常食管上皮细胞HEEC中PHLPP1表达水平。Eca109细胞感染过表达PHLPP1的重组慢病毒(Lv-PHLPP1组),以感染阴性对照重组慢病毒的细胞为Lv-NC组,未行感染的细胞为对照组,qRT-PCR和Western blot法检测3组细胞中PHLPP1的表达,MTT方法测定细胞增殖和黏附能力,Transwell小室检测细胞侵袭和迁移能力,Western blot法检测细胞中增殖细胞核抗原(PCNA)、神经性钙黏附素(N-cadherin)、基质金属蛋白酶1(MMP1)和9(MMP9)、上皮性钙黏附素(E-cadherin)蛋白的表达水平。结果:Eca109、EC9706、KYSE510中PHLPP1表达水平均低于HEEC细胞,Eca109细胞中PHLPP1表达水平最低(P <0. 05)。与对照组或Lv-NC组比较,Lv-PHLPP1组Eca109细胞中PHLPP1 mRNA和蛋白表达水平均升高(P <0. 05),细胞的增殖率、黏附率、侵袭和迁移细胞数均降低,细胞中PCNA、MMP1、MMP9、N-cadherin蛋白表达水平降低,E-cadherin蛋白表达水平升高。结论:PHLPP1在食管癌细胞中表达下调,上调其表达可以抑制食管癌细胞的增殖、黏附、侵袭和迁移能力。
Aim: To study the effects of PHLPP1 on proliferation adhesion,invasion and migration of esophageal cancer cells. Methods: The expressions of PHLPP1 in esophageal carcinoma cells Eca109,EC9706,KYSE510 and normal esophageal epithelial cells HEEC were detected by qRT-PCR and Western blot. Esophageal cancer cells were divided into three groups,the control group was not treated,the Lv-NC group infected with control recombinant lentivirus,and the LvPHLPP1 group infected with PHLPP1 overexpressed recombinant lentivirus. The expression of PHLPP1 was detected by qRT-PCR and Western blot,MTT assay was used to determine cell proliferation and adhesion,the cell invasion and migration ability was detected by Transwell chamber,Western blot was used to detect the expressions of PCNA,MMP1,MMP9,N-cadherin and E-cadherin in the cells. Results: The expression level of PHLPP1 in Eca109,EC9706 and KYSE510 cells was significantly lower than that in HEEC cells,and the expression level of PHLPP1 in Eca109 cells was the lowest(P <0. 05). PHLPP1 mRNA and protein levels in Lv-PHLPP1 group were all elevated. The proliferation,adhesion,invasion and migration of Eca109 cells increased,the levels of PCNA,MMP1,MMP9 and N-cadherin decreased,while the level of E-cadherin protein increased in Lv-PHLPP1 group(P < 0. 05). Conclusion: The expression of PHLPP1 is down-regulated in esophageal cancer cells,and up-regulation of its expression can inhibit the proliferation,adhesion,invasion and migration of esophageal cancer cells.
Aim: To study the effects of PHLPP1 on proliferation adhesion,invasion and migration of esophageal cancer cells. Methods: The expressions of PHLPP1 in esophageal carcinoma cells Eca109,EC9706,KYSE510 and normal esophageal epithelial cells HEEC were detected by qRT-PCR and Western blot. Esophageal cancer cells were divided into three groups,the control group was not treated,the Lv-NC group infected with control recombinant lentivirus,and the LvPHLPP1 group infected with PHLPP1 overexpressed recombinant lentivirus. The expression of PHLPP1 was detected by qRT-PCR and Western blot,MTT assay was used to determine cell proliferation and adhesion,the cell invasion and migration ability was detected by Transwell chamber,Western blot was used to detect the expressions of PCNA,MMP1,MMP9,N-cadherin and E-cadherin in the cells. Results: The expression level of PHLPP1 in Eca109,EC9706 and KYSE510 cells was significantly lower than that in HEEC cells,and the expression level of PHLPP1 in Eca109 cells was the lowest(P <0. 05). PHLPP1 mRNA and protein levels in Lv-PHLPP1 group were all elevated. The proliferation,adhesion,invasion and migration of Eca109 cells increased,the levels of PCNA,MMP1,MMP9 and N-cadherin decreased,while the level of E-cadherin protein increased in Lv-PHLPP1 group(P < 0. 05). Conclusion: The expression of PHLPP1 is down-regulated in esophageal cancer cells,and up-regulation of its expression can inhibit the proliferation,adhesion,invasion and migration of esophageal cancer cells.
引文
[1]GAO T,FURNARI F,NEWTON AC.PHLPP:a phosphatase that directly dephosphorylates Akt,promotes apoptosis,and suppresses tumor growth[J].Mol Cell,2005,18(1):13
[2]冶秀鹏,包慎,李叶琼,等.急性髓系白血病骨髓移植治愈后20年继发食管癌一例[J].白血病·淋巴瘤,2016,25(12):757
[3]HUANG YS,CHANG CC,LEE SS,et al.Xist reduction in breast cancer upregulates AKT phosphorylation via HDAC3-mediated repression of PHLPP1 expression[J].Oncotarget,2016,7(28):43256
[4]AVIV YA.Novel phosphatase PHLPP-1 regulates mitochondrial Akt activity and cardiac cell survival[J].Circ Res,2010,107(4):448
[5]CHEN M,PRATT CP,ZEEMAN ME,et al.Identification of PHLPP1 as a tumor suppressor reveals the role of feedback activation in PTEN-mutant prostate cancer progression[J].Cancer Cell,2011,20(2):173
[6]MEI ZF,HE YC,FENG JJ,et al.MicroRNA-141 promotes the proliferation of non-small cell lung cancer cells by regulating expression of PHLPP1 and PHLPP2[J].FEBSLett,2014,588(17):3055
[7]GAO G,KUN T,SHENG Y,et al.SGT1 regulates Akt signaling by promoting beta-TrCP-dependent PHLPP1 degradation in gastric cancer cells[J].Mol Biol Rep,2013,40(4):2947
[8]庄园.PHLPP1在乳腺癌发生发展中的作用及潜在机制的探讨[D].苏州:苏州大学,2013.
[9]ONO T,NAKAMURA S,KAZUYUKI S,et al.Expression analysis and prognostic significance of the PHLPP gene in acute leukemias and myelodysplastic syndromes[J].Blood,2007,110(11):126
[10]BRADLEY EW,CARPIO LR,WESTENDORF JJ.Histone deacetylase 3 suppression increases PH domain and leucine-rich repeat phosphatase(Phlpp)1 expression in chondrocytes to suppress Akt signaling and matrix secretion[J].J Biol Chem,2013,288(14):9572
[11]LIAO WT,LI TT,WANG ZG,et al.MicroRNA-224 promotes cell proliferation and tumor growth in human colorectal cancer by repressing PHLPP1 and PHLPP2[J].Clin Cancer Res,2013,19(17):4662
[12]ZHOU JY,YU XE,WANG J,et al.Aberrant expression of PHLPP1 and PHLPP2 correlates with poor prognosis in patients with hypopharyngeal squamous cell carcinoma[J].PLo S One,2015,10(3):e0119405
[13]SULJAGIC M,LAURENTI L,TARNANI M,et al.Reduced expression of the tumor suppressor PHLPP1 enhances the antiapoptotic B-cell receptor signal in chronic lymphocytic leukemia B-cells[J].Leukemia,2010,24(12):2063
[14]孙庆敏,王进.miR-200a与金雀异黄酮对胃癌细胞生长及侵袭能力的影响[J].郑州大学学报(医学版),2018,53(4):474
[15]BAI LH,LIN GP,SUN LH,et al.Upregulation of SIRT6predicts poor prognosis and promotes metastasis of nonsmall cell lung cancer via the ERK1/2/MMP9 pathway[J].Oncotarget,2016,7(26):40377
[16]林锋,王海娟,李春晓,等.食管癌细胞来源的外泌体对肿瘤细胞迁移及侵袭能力的影响及机制研究[J].解放军医学杂志,2017,42(4):307
[17]LI D,WEI Y,WANG D,et al.MicroRNA-26b suppresses the metastasis of non-small cell lung cancer by targeting MIEN1 via NF-κB/MMP-9/VEGF pathways[J].Biochem Biophys Res Commun,2016,472(3):465
[18]LIU YS,SUN XJ,FENG JF,et al.MT2-MMP induces proteolysis and leads to EMT in carcinomas[J].Oncotarget,2016,7(30):48193
[19]BISWAS S,SENGUPTA S,ROY CHOWDHURY S,et al.Erratum to:CXCL13-CXCR5 co-expression regulates epithelial to mesenchymal transition of breast cancer cells during lymph node metastasis[J].Breast Cancer Res Treat,2016,155(3):615
[20]YAO L,ZHANG D,ZHAO X,et al.Dickkopf-1-promoted vasculogenic mimicry in non-small cell lung cancer is associated with EMT and development of a cancer stem-like cell phenotype[J].J Cell Mol Med,2016,20(9):1673
[2]冶秀鹏,包慎,李叶琼,等.急性髓系白血病骨髓移植治愈后20年继发食管癌一例[J].白血病·淋巴瘤,2016,25(12):757
[3]HUANG YS,CHANG CC,LEE SS,et al.Xist reduction in breast cancer upregulates AKT phosphorylation via HDAC3-mediated repression of PHLPP1 expression[J].Oncotarget,2016,7(28):43256
[4]AVIV YA.Novel phosphatase PHLPP-1 regulates mitochondrial Akt activity and cardiac cell survival[J].Circ Res,2010,107(4):448
[5]CHEN M,PRATT CP,ZEEMAN ME,et al.Identification of PHLPP1 as a tumor suppressor reveals the role of feedback activation in PTEN-mutant prostate cancer progression[J].Cancer Cell,2011,20(2):173
[6]MEI ZF,HE YC,FENG JJ,et al.MicroRNA-141 promotes the proliferation of non-small cell lung cancer cells by regulating expression of PHLPP1 and PHLPP2[J].FEBSLett,2014,588(17):3055
[7]GAO G,KUN T,SHENG Y,et al.SGT1 regulates Akt signaling by promoting beta-TrCP-dependent PHLPP1 degradation in gastric cancer cells[J].Mol Biol Rep,2013,40(4):2947
[8]庄园.PHLPP1在乳腺癌发生发展中的作用及潜在机制的探讨[D].苏州:苏州大学,2013.
[9]ONO T,NAKAMURA S,KAZUYUKI S,et al.Expression analysis and prognostic significance of the PHLPP gene in acute leukemias and myelodysplastic syndromes[J].Blood,2007,110(11):126
[10]BRADLEY EW,CARPIO LR,WESTENDORF JJ.Histone deacetylase 3 suppression increases PH domain and leucine-rich repeat phosphatase(Phlpp)1 expression in chondrocytes to suppress Akt signaling and matrix secretion[J].J Biol Chem,2013,288(14):9572
[11]LIAO WT,LI TT,WANG ZG,et al.MicroRNA-224 promotes cell proliferation and tumor growth in human colorectal cancer by repressing PHLPP1 and PHLPP2[J].Clin Cancer Res,2013,19(17):4662
[12]ZHOU JY,YU XE,WANG J,et al.Aberrant expression of PHLPP1 and PHLPP2 correlates with poor prognosis in patients with hypopharyngeal squamous cell carcinoma[J].PLo S One,2015,10(3):e0119405
[13]SULJAGIC M,LAURENTI L,TARNANI M,et al.Reduced expression of the tumor suppressor PHLPP1 enhances the antiapoptotic B-cell receptor signal in chronic lymphocytic leukemia B-cells[J].Leukemia,2010,24(12):2063
[14]孙庆敏,王进.miR-200a与金雀异黄酮对胃癌细胞生长及侵袭能力的影响[J].郑州大学学报(医学版),2018,53(4):474
[15]BAI LH,LIN GP,SUN LH,et al.Upregulation of SIRT6predicts poor prognosis and promotes metastasis of nonsmall cell lung cancer via the ERK1/2/MMP9 pathway[J].Oncotarget,2016,7(26):40377
[16]林锋,王海娟,李春晓,等.食管癌细胞来源的外泌体对肿瘤细胞迁移及侵袭能力的影响及机制研究[J].解放军医学杂志,2017,42(4):307
[17]LI D,WEI Y,WANG D,et al.MicroRNA-26b suppresses the metastasis of non-small cell lung cancer by targeting MIEN1 via NF-κB/MMP-9/VEGF pathways[J].Biochem Biophys Res Commun,2016,472(3):465
[18]LIU YS,SUN XJ,FENG JF,et al.MT2-MMP induces proteolysis and leads to EMT in carcinomas[J].Oncotarget,2016,7(30):48193
[19]BISWAS S,SENGUPTA S,ROY CHOWDHURY S,et al.Erratum to:CXCL13-CXCR5 co-expression regulates epithelial to mesenchymal transition of breast cancer cells during lymph node metastasis[J].Breast Cancer Res Treat,2016,155(3):615
[20]YAO L,ZHANG D,ZHAO X,et al.Dickkopf-1-promoted vasculogenic mimicry in non-small cell lung cancer is associated with EMT and development of a cancer stem-like cell phenotype[J].J Cell Mol Med,2016,20(9):1673