Beclin-1在稽留流产中的表达模式及临床意义
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摘要
目的自噬在稽留流产中的相关研究鲜有报道。文中通过检测自噬相关因子Beclin-1及LC3在正常妊娠者和稽留流产者的绒毛及蜕膜中的表达情况,探讨稽留流产与细胞自噬之间的关系,为稽留流产的早期防治提供理论依据。方法选取2017年1月至3月在江苏大学第四附属医院妇科收治的30例稽留流产患者作为稽留组,同期收治的30例正常妊娠患者作为对照组。利用荧光定量PCR方法检测2组绒毛及蜕膜组织中Beclin-1 mRNA的表达水平,利用Western Blot检测两组绒毛及蜕膜组织中Beclin-1、LC3II/LC3I蛋白的表达水平,利用免疫组化观察Beclin-1在2组绒毛及蜕膜组织中的定位情况。结果与对照组相比,Beclin-1 mRNA在稽留流产绒毛(1.00±0.28 vs 2.17±0.87)及蜕膜组织(1.75±0.41 vs 4.74±0.93)中显著性上调,差异有统计学意义(P<0.05)。Beclin-1和LC3Ⅱ/LC3Ⅰ蛋白在稽留流产绒毛及蜕膜组织中均呈上调趋势,差异有统计学意义(P<0.05)。免疫组化显示Beclin-1蛋白在稽留流产绒毛及蜕膜组织中的表达水平均呈现上调趋势。结论Beclin-1 mRNA及蛋白、LC3Ⅱ/LC3Ⅰ在稽留流产的绒毛及蜕膜组织中均上调,提示细胞自噬参与了稽留流产的发生,为研究稽留流产的发病机制提供新的理论依据。
Objective Reports are rarely seen on the role of autophagy in missed abortion. This study aimed to explore the association of missed abortion with autophagy by detecting the expressions of the autophagy-related factors Beclin-1 and LC3 in the villus and decidua of normal pregnancy and missed abortion women so as to provide some theoretical evidence for the early prevention of missed abortion. Methods We included in the experimental group 30 missed abortion patients treated in the Department of Obstetrics and Gynecology,the Fourth Affiliated Hospital of Jiangsu University,from January to March 2017 and enrolled another 30 normal pregnancy women as controls. We determined the expression level of Beclin-1 mRNA in the villus and decidua by quantitative fluorescence PCR and those of Beclin-1 and LC3 II/LC3 I proteins by Western blot. We also positioned Beclin-1 in the villus and decidua tissues of the two groups of women by immunohistochemistry. Results Compared with the normal pregnancy women,the patients with missed abortion showed significantly up-regulated expression of Beclin-1 mRNA in the villus( 1.00±0.28 vs 2.17±0.87,P<0.05) and decidua( 1.75±0.41 vs 4.74±0.93,P< 0.05) and those of Beclin-1 and LC3 II/LC3 I proteins( P< 0.05). Immunohistochemistry revealed markedly increased expression of the Beclin-1 protein in the villus and decidua of the missed abortion patients( P< 0.05).Conclusion Beclin-1 mRNA,Beclin-1 protein and LC3 II/LC3 I are all up-regulated in the villus and decidua of missed abortion patients,suggesting the involvement of autophagy in the occurrence of missed abortion,which may provide a new theoretical basis for studying the pathogenesis of missed abortion.
Objective Reports are rarely seen on the role of autophagy in missed abortion. This study aimed to explore the association of missed abortion with autophagy by detecting the expressions of the autophagy-related factors Beclin-1 and LC3 in the villus and decidua of normal pregnancy and missed abortion women so as to provide some theoretical evidence for the early prevention of missed abortion. Methods We included in the experimental group 30 missed abortion patients treated in the Department of Obstetrics and Gynecology,the Fourth Affiliated Hospital of Jiangsu University,from January to March 2017 and enrolled another 30 normal pregnancy women as controls. We determined the expression level of Beclin-1 mRNA in the villus and decidua by quantitative fluorescence PCR and those of Beclin-1 and LC3 II/LC3 I proteins by Western blot. We also positioned Beclin-1 in the villus and decidua tissues of the two groups of women by immunohistochemistry. Results Compared with the normal pregnancy women,the patients with missed abortion showed significantly up-regulated expression of Beclin-1 mRNA in the villus( 1.00±0.28 vs 2.17±0.87,P<0.05) and decidua( 1.75±0.41 vs 4.74±0.93,P< 0.05) and those of Beclin-1 and LC3 II/LC3 I proteins( P< 0.05). Immunohistochemistry revealed markedly increased expression of the Beclin-1 protein in the villus and decidua of the missed abortion patients( P< 0.05).Conclusion Beclin-1 mRNA,Beclin-1 protein and LC3 II/LC3 I are all up-regulated in the villus and decidua of missed abortion patients,suggesting the involvement of autophagy in the occurrence of missed abortion,which may provide a new theoretical basis for studying the pathogenesis of missed abortion.
引文
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[10]Gao L,Qi HB,Kc K,et al.Excessive autophagy induces the failure of trophoblast invasion and vasculature:possible relevance to the pathogenesis of preeclampsia[J].J Hypert,2015,33(1):106-117.
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[12]Mizushima N,Levine B,Cuervo AM,et al.Autophagy fights disease through cellular self-digestion[J].Nature,2008,451(7182):1069.
[13]Yang Z,Klionsky DJ.Eaten alive:a history of macroautophagy[J].Nat Cell Biol,2010,12(9):814.
[14]He C,Levine B.The Beclin 1 interactome[J].Curr Opin Cell Biol,2010,22(2):140-149.
[15]Oh SY,Choi SJ,Kim KH,et al.Autophagy-related proteins,LC3 and Beclin-1,in placentas from pregnancies complicated by preeclampsia[J].Rep Sci,2008,15(9):912-920.
[16]Su Z,Yang Z,Xu Y,et al.Apoptosis,autophagy,necroptosis,and cancer metastasis[J].Mol Cancer,2015,14(1):48.
[17]Gorski SM,Chittaranjan S,Pleasance ED,et al.A SAGE Approach to Discovery of Genes Involved in Autophagic Cell Death[J].Curr Biol,2003,13(4):358-363.
[18]Zhi Z,Yang W,Liu L,et al.Early missed abortion is associated with villous angiogenesis via the HIF-1α/VEGF signaling pathway[J].Arch Gynecol Obstet,2018,298(3):537-543.
[2]邹琳,何红华,庞小艳,等.低分子肝素在人类辅助生殖技术获得妊娠后发生复发性流产患者中的临床研究[J].医学研究生学报,2017,30(4):399-404.
[3]刘芳.IL-6及Survivin在胚胎停育中的临床意义[J].中国妇幼保健,2015,30(22):3882-3884.
[4]Xia S,Zhen Y,Ma H,et al.Abnormal expression of microRNA-575 leads to missed abortion through regulating apoptosis and angiogenesis[J].Exp Ther Med,2017,14(5):3993.
[5]苑杰,江沛,吕佩源,等.细胞自噬与疾病发生发展的关系及作为治疗靶点的研究进展[J].医学研究生学报,2018,31(1):88-91.
[6]Chen B,Longtine MS,Nelson DM.Hypoxia induces autophagy in primary human trophoblasts[J].Endocrinology,2012,153(10):4946-4954.
[7]Barbara C,Maria Teresa L,Gloria L,et al.Autophagy-related protein LC3 and Beclin-1 in the first trimester of pregnancy[J].Clin Exp Rep Med,2013,40(1):33.
[8]Katayama M,Kawaguchi TM,Pieper R.DNA damaging agentinduced autophagy produces a cytoprotective adenosine triphosphate surge in malignant glioma cells[J].Cell Death Differ,2007,14(4):548-558.
[9]Yang ZJ,Chee CE,Huang S,et al.The role of autophagy in cancer:therapeutic implications[J].Mol Cancer Ther,2011,10(9):1533-1541.
[10]Gao L,Qi HB,Kc K,et al.Excessive autophagy induces the failure of trophoblast invasion and vasculature:possible relevance to the pathogenesis of preeclampsia[J].J Hypert,2015,33(1):106-117.
[11]Agrawal V,Jaiswal MK,Mallers T,et al.Altered autophagic flux enhances inflammatory responses during inflammation-induced preterm labor[J].Sci Rep,2015,5:9410.
[12]Mizushima N,Levine B,Cuervo AM,et al.Autophagy fights disease through cellular self-digestion[J].Nature,2008,451(7182):1069.
[13]Yang Z,Klionsky DJ.Eaten alive:a history of macroautophagy[J].Nat Cell Biol,2010,12(9):814.
[14]He C,Levine B.The Beclin 1 interactome[J].Curr Opin Cell Biol,2010,22(2):140-149.
[15]Oh SY,Choi SJ,Kim KH,et al.Autophagy-related proteins,LC3 and Beclin-1,in placentas from pregnancies complicated by preeclampsia[J].Rep Sci,2008,15(9):912-920.
[16]Su Z,Yang Z,Xu Y,et al.Apoptosis,autophagy,necroptosis,and cancer metastasis[J].Mol Cancer,2015,14(1):48.
[17]Gorski SM,Chittaranjan S,Pleasance ED,et al.A SAGE Approach to Discovery of Genes Involved in Autophagic Cell Death[J].Curr Biol,2003,13(4):358-363.
[18]Zhi Z,Yang W,Liu L,et al.Early missed abortion is associated with villous angiogenesis via the HIF-1α/VEGF signaling pathway[J].Arch Gynecol Obstet,2018,298(3):537-543.