Effect and mechanism of the aβ2‑GP I/rhβ2‑GP I complex on JEG‑3 cell proliferation, migration and invasion

Lu,Xiumin; Ren,Lei; Zhang,Wenjing; Liu,Yanhong

doi:10.3892/mmr.2018.8822

Effect and mechanism of the aβ2‑GP I/rhβ2‑GP I complex on JEG‑3 cell proliferation, migration and invasion

Authors:
- Xiumin Lu
- Lei Ren
- Wenjing Zhang
- Yanhong Liu
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Affiliations: Departments of Prenatal Diagnosis, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Departments of Laboratory Diagnosis, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: March 29, 2018 https://doi.org/10.3892/mmr.2018.8822
Pages: 7505-7512
Copyright: © Lu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Antiphospholipid antibody (aPL)‑mediated antiphospholipid syndrome (APS) is an autoimmune disease. Upon binding to aPL, the primary antigen of aPL, β2‑glycoprotein I (β2‑GP I), induces abnormal immune function, which further activates downstream signaling pathways in the cell and eventually leads to APS. The present study aimed to determine whether β2‑GP I antigen and anti‑β2‑glycoprotein I antibody (aβ2‑GP I), which belong to the aPL class of antibodies, may affect human chorionic epithelium cell (JEG‑3) proliferation, migration and invasion. Recombinant human (rh)β2‑GP I protein was expressed using a prokaryotic expression system and aβ2‑GP I antibody was purified from the blood serum of 10 patients with recurrent pregnancy loss. JEG‑3 cells were stimulated with rhβ2‑GP I and aβ2‑GP I separately or simultaneously, and serum immunoglobulin G of normal pregnant women was used as negative control. Using cell counting kit‑8, cell cycle and transwell assays in addition to EdU staining, it was determined that aβ2‑GP I/rhβ2‑GP I complex markedly increased JEG‑3 cell proliferation, migration and invasion. The results revealed that mRNA levels of inhibitor of nuclear factor (NF)‑κB kinase subunit (IKKβ), myeloid differentiation primary response protein MyD88 (MyD88), NF‑κB and NF‑κB inhibitor α (IκBα), as well as the protein levels of MyD88, IκBα and phospho(p)‑IκBα in JEG‑3 cells increased following incubation with the aβ2‑GP I/rhβ2‑GP I complex. The observed upregulation of p‑IκBα protein suggested that IκBα‑mediated inhibition of NF‑κB was weakened. Furthermore, JEG‑3 cells were transfected with PGMLV‑NF‑κB‑Lu vector. Luciferase activity in JEG‑3‑NFκB‑Luc1 and JEG‑3‑NFκB‑Luc2 cells was enhanced following treatment with aβ2‑GP I/rhβ2‑GP I complex. The present study demonstrated that aβ2‑GP I/rhβ2‑GP I complex activates NF‑κB through MyD88 signal transduction pathway, which further enhances JEG‑3 cell proliferation, migration and invasion.

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1	Lee NS, Brewer HB Jr and Osborne JC Jr: Beta 2-Glycoprotien I: Molecular properties of an unusual apolipoprotein H. J Biol Chem. 258:4765–4770. 1983.PubMed/NCBI
2	Rand JH, Wu XX, Andree HA, Ross JB, Rusinova E, Gascon-Lema MG, Calandri C and Harpel PC: Antiphospholipid antibodies accelerate plasma coagulation by inhibiting annexin-V binding to phospholipids: A ‘Lupus Procoagulant’ phenomenon. Blood. 92:1652–1660. 1998.PubMed/NCBI
3	Pittoni V, Ravirajan CT, Donohoe S, Machin SJ, Lydyard PM and Isenberg DA: Human monoclonal anti-phospholipid antibodies selectively bind to membrane phospholipid and beta2-glycoprotein I (beta2-GPI) on apoptotic cells. Clin Exp Immunol. 119:533–543. 2000. View Article : Google Scholar : PubMed/NCBI
4	Gao PJ, Shi Y, Gao YH, Liu YW and Tan Y: The receptor for beta(2)GP I on membrane of hepatocellular carcinoma cell line SMMC-7721 is annexin II. World J Gastroenterol. 13:3364–3368. 2007. View Article : Google Scholar : PubMed/NCBI
5	Miyakis S, Lockshin MD, Atsumi T, Branch DW, Brey RL, Cervera R, Derksen RH, DE Groot PG, Koike T, Meroni PL, et al: International consensus statement on an update of the classification criteria for definite antiphospholipid syndrome (APS). J Thromb Haemost. 4:295–306. 2006. View Article : Google Scholar : PubMed/NCBI
6	Pablo Demetrio R, Muñoz P, Hoyos López M, Calvo V, Riancho L and Taboada Martinez VM: Thrombocytopenia as a thrombotic risk factor in patients with antiphospholipid antibodies without disease criteria. Med Clin (Barc). 148:394–400. 2017.PubMed/NCBI
7	Cris JC, Bouvier S, Nouvellon E, Lissalde-Lavigne G, Mercier E, Balducchi JP and Marès P: Antiphospholid antibodies and the risk of pregnancy complication. Thromb Res. 151 Suppl 1:S34–S37. 2017. View Article : Google Scholar : PubMed/NCBI
8	Chighizola CB, Raimondo MG and Meroni PL: Management of thrombotic antiphospholipid syndrome. Semin Thromb Hemost. Mar 9–2017.(Epub ahead of print). View Article : Google Scholar : PubMed/NCBI
9	Kandiah DA, Sheng YH and Krilis SA: beta 2-Glycoprotein I: Target antigen for autoantibodies in the ‘antiphospholipid syndrome. Lupus. 5:381–385. 1996. View Article : Google Scholar : PubMed/NCBI
10	Dalekos GN, Zachou K and Liaskos C: The antiphospholipid syndrome and infection. Curr Rheumatol Rep. 3:277–285. 2001. View Article : Google Scholar : PubMed/NCBI
11	Raschi E, Chighizola CB, Grossi C, Ronda N, Gatti R, Meroni PL and Borghi MO: β2-glycoprotein I, lipopolysaccharide and endothelial TLR4: Three players in the two hit theory for anti-phospholipid-mediated thrombosis. J Autoimmun. 55:42–50. 2014. View Article : Google Scholar : PubMed/NCBI
12	Robertson SA, Roberts CT, van Beijering E, Pensa K, Sheng Y, Shi T and Krilis SA: Effect of beta2-glycoprotein I null mutation on reproductive outcome and antiphospholipid antibody-mediated pregnancy pathology in mice. Mol Hum Reprod. 10:409–416. 2004. View Article : Google Scholar : PubMed/NCBI
13	Steinkasserer A, Cockburn DJ, Black DM, Boyd Y, Solomon E and Sim RB: Assignment of apolipoprotein H (APOH: beta-2-glycoprotein I) to human chromosome 17q23----qter; determination of the major expression site. Cytogenet Cell Genet. 60:31–33. 1992. View Article : Google Scholar : PubMed/NCBI
14	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
15	Hernández-Ramirez DF, Olivares-Martinez E, Núñez-Álvarez CA, Chavelas EA, Garcia-Hernandez E, Gómez-Hernandez G, Llorente L and Cabral AR: The role of β2-glycoprotein I (β2GPI) carbohydrate chains in the reactivity of anti-β2GPI antibodies from patients with primary antiphospholipid syndrome and in the activation and differentiation of U937 cells. Biochem Biophys Res Commun. 453:94–100. 2014. View Article : Google Scholar : PubMed/NCBI
16	Muzio M, Polentarutti N, Bosisio D, Kumar Manoj PP and Mantovani A: Toll-like receptor family and signalling pathway. Biochem Soc Trans. 28:563–566. 2000. View Article : Google Scholar : PubMed/NCBI
17	Gyrd-Hansen M and Meier P: IAPs: From caspase inhibitors to modulators of NF-kappaB, inflammation and cancer. Nat Rev Cancer. 10:561–574. 2010. View Article : Google Scholar : PubMed/NCBI
18	Xie H, Zhou H, Wang H, Chen D, Xia L, Wang T and Yan J: Anti-β(2)GPI/β(2)GPI induced TF and TNF-α expression in monocytes involving both TLR4/MyD88 and TLR4/TRIF signaling pathways. Mol Immunol. 53:246–254. 2013. View Article : Google Scholar : PubMed/NCBI
19	De Laat B, Derksen RH, van Lummel M, Pennings MT and de Groot PG: Pathogenic anti-beta2-glycoprotein I antibodies recognize domain I of beta2-glycoprotein I only after a conformational change. Blood. 107:1916–1924. 2006. View Article : Google Scholar : PubMed/NCBI
20	Lutters BC, Derksen RH, Tekelenburg WL, Lenting PJ, Arnout J and de Groot PG: Dimers of beta 2-glycoprotein I increase platelet deposition to collagen via interaction with phospholipids and the apolipoprotein E receptor 2. J Biol Chem. 278:33831–33838. 2003. View Article : Google Scholar : PubMed/NCBI
21	De Groot PG and Meijers JC: β(2)-glycoprotein I: Evolution, structure and function. J Thromb Haemost. 9:1275–1284. 2011. View Article : Google Scholar : PubMed/NCBI
22	Ailus K, Tulppala M, Palosuo T, Ylikorkala O and Vaarala O: Antibodies to beta 2-glycoprotein I and prothrombin in habitual abortion. Fertil Steril. 66:937–941. 1996. View Article : Google Scholar : PubMed/NCBI
23	Vinatier D, Dufour P, Cosson M and Houpeau JL: Antiphospholipid syndrome and recurrent miscarriages. Eur Obstet Gynecol Reproduct Biol. 96:37–50. 2001. View Article : Google Scholar
24	Sada PR, Cohen H and Isenberg D: The pathophysiology of antiphospholipid syndrome. Open Urol Nephrol J. 8:2–9. 2015. View Article : Google Scholar
25	Alijotas-Reig J, Esteve-Valverde E, Ferrer-Oliveras R, Llurba E and Gris JM: Tumor necrosis factor-alpha and pregnancy: Focus on biologics. An updated and comprehensive review. Clin Rev Allergy Immunol. 53:40–53. 2017. View Article : Google Scholar : PubMed/NCBI
26	Weiss R, Bitton A, Nahary L, Arango MT, Benhar I, Blank M, Shoenfeld Y and Chapman J: Cross-reactivity between annexin A2 and Beta-2-glycoprotein I in animal models of antiphospholipid syndrome. Immunol Res. 65:355–362. 2017. View Article : Google Scholar : PubMed/NCBI
27	Yoshimura A: Signal transduction of inflammatory cytokines and tumor development. Cancer Sci. 97:439–447. 2006. View Article : Google Scholar : PubMed/NCBI
28	Ball E, Bulmer JN, Ayis S, Lyall F and Robson SC: Late sporadic miscarriage is associated with abnormalities in spiral artery transformation and trophoblast invasion. J Pathol. 208:535–542. 2006. View Article : Google Scholar : PubMed/NCBI
29	Ferretti C, Bruni L, Dangles-Marie V, Pecking AP and Bellet D: Molecular circuits shared by placental and cancer cells, and their implications in the proliferative, invasive and migratory capacities of trophoblasts. Hum Reprod Update. 13:121–141. 2007. View Article : Google Scholar : PubMed/NCBI
30	Aardema MW, Oosterhof H, Timmer A, van Rooy I and Aarnoudse JG: Uterine artery Doppler flow and uteroplacental vascular pathology in normal pregnancies and pregnancies complicated by pre-eclampsia and small for gestational age fetuses. Placenta. 22:405–411. 2001. View Article : Google Scholar : PubMed/NCBI
31	Kim YM, Chaiworapongsa T, Gomez R, Bujold E, Yoon BH, Rotmensch S, Thaler HT and Romero R: Failure of physiologic transformation of the spiral arteries in the placental bed in preterm premature rupture of membranes. Am J Obstet Gynecol. 187:1137–1142. 2002. View Article : Google Scholar : PubMed/NCBI
32	Kim YM, Bujold E, Chaiworapongsa T, Gomez R, Yoon BH, Thaler HT, Rotmensch S and Romero R: Failure of physiologic transformation of the spiral arteries in patients with preterm labor and intact membranes. Am J Obstet Gynecol. 189:1063–1069. 2003. View Article : Google Scholar : PubMed/NCBI
33	Cui JQ, Shi YF, Zhou HJ and Li JQ: Changes in gene expression profiles of hydatidiform mole and choriocarcinoma as compared with trophoblast hyperplasia. Zhonghua Zhong Liu Za Zhi. 26:727–731. 2004.(In Chinese). PubMed/NCBI