Fibronectin 1 inhibits the apoptosis of human trophoblasts by activating the PI3K/Akt signaling pathway

Ji,Jinlong; Chen,Liping; Zhuang,Yanyan; Han,Yun; Tang,Weichun; Xia,Fei

doi:10.3892/ijmm.2020.4735

Fibronectin 1 inhibits the apoptosis of human trophoblasts by activating the PI3K/Akt signaling pathway

Authors:
- Jinlong Ji
- Liping Chen
- Yanyan Zhuang
- Yun Han
- Weichun Tang
- Fei Xia
View Affiliations

Affiliations: Department of Gynecology and Obstetrics, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China, Department of Gynecology and Obstetrics, The Second Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
Published online on: September 23, 2020 https://doi.org/10.3892/ijmm.2020.4735
Pages: 1908-1922
Copyright: © Ji et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The excessive apoptosis of human trophoblasts can cause pregnancy‑related diseases. It has been reported that fibronectin 1 (FN1) is closely associated with the invasion of human trophoblasts. The aim of the present study was to examine the effects of FN1 on the apoptosis of human trophoblasts and to investigate the underlying molecular mechanisms. It was found that FN1, a differentially expressed gene (DEG) in the GSE127170 dataset, was identified as the hub gene in a protein‑protein interaction (PPI) network generated using the cytoHubba plug‑in of Cytoscape software. The Metascape website was used to perform GO enrichment analysis, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database was used to perform KEGG pathway analysis. Experimental analyses revealed that FN1 expression was downregulated in the chorionic villus tissues of patients diagnosed with and mice subjected to spontaneous abortion (SA). CCK‑8 and flow cytometric assays revealed that the knockdown of FN1 decreased the viability and promoted the apoptosis of JEG‑3 and BeWo cells. In vivo experiments demonstrated that the knockdown of FN1 promoted the apoptosis of trophoblasts in the chorionic villus tissues obtained from mice subjected to SA, whereas FN1 overexpression increased cell viability and inhibited cell apoptosis. The protein levels of cleaved caspase‑3 and Bax were increased by the silencing of FN1 and decreased by FN1 overexpression. The protein expression levels of Bcl‑2, proliferating cell nuclear antigen (PCNA) and Ki67 were decreased by the silencing of FN1; however, the overexpression of FN1 increased these levels. The results of western blot analysis revealed that the knockdown of FN1 inhibited the PI3K/Akt signaling pathway, while the overexpression of FN1 activated the PI3K/Akt signaling pathway. Consistently, the apoptosis‑inhibiting effect of FN1 overexpression was reversed by a PI3K/Akt signaling pathway inhibitor, and the apoptosis‑promoting effect of FN1 silencing was reversed by a PI3K/Akt signaling pathway activator. On the whole, the findings of the present study demonstrate that the inhibition of FN1 induces the apoptosis of JEG‑3 and BeWo cells, and the overexpression of FN1 inhibits cell apoptosis by activating the PI3K/Akt signaling pathway.

View Figures

View References

1	Moser G, Windsperger K, Pollheimer J, de Sousa Lopes SC and Huppertz B: Human trophoblast invasion: New and unexpected routes and functions. Histochem Cell Biol. 150:361–370. 2018. View Article : Google Scholar : PubMed/NCBI
2	Takahashi H, Ogoyama M, Nagayama S, Suzuki H, Ohkuchi A, Matsubara S and Takizawa T: Extravillous trophoblast invasion accelerated by WNT3A, 5A, and 10B via CD44. J Matern Fetal Neonatal Med. 1–9. 2019.PubMed/NCBI
3	Walentin K, Hinze C and Schmidt-Ott KM: The basal chorionic trophoblast cell layer: An emerging coordinator of placenta development. Bioessays. 38:254–265. 2016. View Article : Google Scholar : PubMed/NCBI
4	Ohgaki R, Ohmori T, Hara S, Nakagomi S, Kanai-Azuma M, Kaneda-Nakashima K, Okuda S, Nagamori S and Kanai Y: Essential roles of L-Type amino acid transporter 1 in syncytiotrophoblast development by presenting fusogenic 4F2hc. Mol Cell Biol. 37:e00427–16. 2017. View Article : Google Scholar : PubMed/NCBI
5	Gu Y, Zhao S, Wan J, Meng J, Zuo C, Wang S, Zhou Y, Li H and Wang X: Hsa-miRNA-125b may induce apoptosis of HTR8/SVneo cells by targeting MCL1. Reprod Biol. 19:368–373. 2019. View Article : Google Scholar : PubMed/NCBI
6	Ding J, Yin T, Yan N, Cheng Y and Yang J: FasL on decidual macrophages mediates trophoblast apoptosis: A potential cause of recurrent miscarriage. Int J Mol Med. 43:2376–2386. 2019.PubMed/NCBI
7	Liu HN, Tang XM, Wang XQ, Gao J, Li N, Wang YY and Xia HF: MiR-93 inhibits trophoblast cell proliferation and promotes cell apoptosis by targeting BCL2L2 in recurrent spontaneous abortion. Reprod Sci. 27:152–162. 2020. View Article : Google Scholar : PubMed/NCBI
8	Tian S, Yu J, Zhang Y, Bian Y, Ma J and Yan J: Overexpression of PTEN regulated by miR-19b and miR-494 in the villous of recurrent spontaneous abortion patients. J Reprod Immunol. 140:1031332020. View Article : Google Scholar : PubMed/NCBI
9	Yang D, Ding J, Wang Y, Yuan M, Xian S, Zhang L, Liu S, Dai D, Wang F, Zheng Y, et al: YY1-PVT1 affects trophoblast invasion and adhesion by regulating mTOR pathway-mediated autophagy. J Cell Physiol. 235:6637–6646. 2020. View Article : Google Scholar : PubMed/NCBI
10	Xu TP, Huang MD, Xia R, Liu XX, Sun M, Yin L, Chen WM, Han L, Zhang EB, Kong R, et al: Decreased expression of the long non-coding RNA FENDRR is associated with poor prognosis in gastric cancer and FENDRR regulates gastric cancer cell metastasis by affecting fibronectin1 expression. J Hematol Oncol. 7:632014. View Article : Google Scholar : PubMed/NCBI
11	Chen Z, Tao Q, Qiao B and Zhang L: Silencing of LINC01116 suppresses the development of oral squamous cell carcinoma by up-regulating microRNA-136 to inhibit FN1. Cancer Manag Res. 11:6043–6059. 2019. View Article : Google Scholar : PubMed/NCBI
12	Dong Z, Lin C, Liu Y, Jin H, Wu H, Li Z, Sun L, Zhang L, Hu X, Wei Y, et al: Upregulation of sestrins protect atriums against oxidative damage and fibrosis in human and experimental atrial fibrillation. Sci Rep. 7:463072017. View Article : Google Scholar : PubMed/NCBI
13	Satoh A, Niwano S, Niwano H, Kishihara J, Aoyama Y, Oikawa J, Fukaya H, Tamaki H and Ako J: Aliskiren suppresses atrial electrical and structural remodeling in a canine model of atrial fibrillation. Heart Vessels. 32:90–100. 2017. View Article : Google Scholar
14	Tian L, Lu ZP, Cai BB, Zhao LT, Qian D, Xu QC, Wu PF, Zhu Y, Zhang JJ, Du Q, et al: Activation of pancreatic stellate cells involves an EMT-like process. Int J Oncol. 48:783–792. 2016. View Article : Google Scholar
15	Wang Y, Fu Y, Yan Z, Zhang XB and Pei M: Impact of fibronectin knockout on proliferation and differentiation of human infrapatellar fat pad-derived stem cells. Front Bioeng Biotechnol. 7:3212019. View Article : Google Scholar : PubMed/NCBI
16	Hou J, Li L, Zhu H, Chen H, Wei N, Dai M, Ni Q and Guo X: Association between breast cancer cell migration and radiosensitivity in vitro. Oncol Lett. 18:6877–6884. 2019.PubMed/NCBI
17	Gao R, Feng Q and Tan G: microRNA-613 exerts anti-angiogenic effect on nasopharyngeal carcinoma cells through inactivating the AKT signaling pathway by down-regulating FN1. Biosci Rep. 39:BSR201821962019. View Article : Google Scholar : PubMed/NCBI
18	Zhang S, Wang Q, Han Q, Han H and Lu P: Identification and analysis of genes associated with papillary thyroid carcinoma by bioinformatics methods. Biosci Rep. 39:BSR201900832019. View Article : Google Scholar : PubMed/NCBI
19	Litovkin KV, Ivanova OV, Bauer A, Hoheisel JD, Bubnov VV and Zaporozhan VN: Microarray study of gene expression in uterine leiomyoma. Exp Oncol. 30:106–111. 2008.PubMed/NCBI
20	Mu Y, Zhou DN, Yan NN, Ding JL and Yang J: Upregulation of ADAMTS7 and downregulation of COMP are associated with spontaneous abortion. Mol Med Rep. 19:2620–2626. 2019.PubMed/NCBI
21	Lu X, Cui J, Cui L, Luo Q, Cao Q, Yuan W and Zhang H: The effects of human umbilical cord-derived mesenchymal stem cell transplantation on endometrial receptivity are associated with Th1/Th2 balance change and uNK cell expression of uterine in autoimmune premature ovarian failure mice. Stem Cell Res Ther. 10:2142019. View Article : Google Scholar : PubMed/NCBI
22	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
23	Zipori Y, Haas J, Berger H and Barzilay E: Multifetal pregnancy reduction of triplets to twins compared with non-reduced triplets: A meta-analysis. Reprod Biomed Online. 35:296–304. 2017. View Article : Google Scholar : PubMed/NCBI
24	Lorek D, Kedzierska AE, Slawek A and Chelmonska-Soyta A: Expression of Toll-like receptors and costimulatory molecules in splenic B cells in a normal and abortion-prone murine pregnancy model. Am J Reprod Immunol. 82:e131482019. View Article : Google Scholar : PubMed/NCBI
25	Xu WM, Xiao ZN, Wang XB and Huang Y: IL-17 induces fetal loss in a CBA/JxBALB/c mouse model, and an anti-IL-17 antibody prevents fetal loss in a CBA/JxDBA/2 mouse model. Am J Reprod Immunol. 75:51–58. 2016. View Article : Google Scholar
26	Ciaramella V, Sasso FC, Di Liello R, Corte CM, Barra G, Viscardi G, Esposito G, Sparano F, Troiani T, Martinelli E, et al: Activity and molecular targets of pioglitazone via blockade of proliferation, invasiveness and bioenergetics in human NSCLC. J Exp Clin Cancer Res. 38:1782019. View Article : Google Scholar : PubMed/NCBI
27	Moon JR, Oh SJ, Lee CK, Chi SG and Kim HJ: TGF-β1 protects colon tumor cells from apoptosis through XAF1 suppression. Int J Oncol. 54:2117–2126. 2019.PubMed/NCBI
28	Tang RZ, Gu SS, Chen XT, He LJ, Wang KP and Liu XQ: Immobilized transforming growth factor-Beta 1 in a stiffness-tunable artificial extracellular matrix enhances mechanotransduction in the epithelial mesenchymal transition of hepatocellular carcinoma. ACS Appl Mater Interfaces. 11:14660–14671. 2019. View Article : Google Scholar : PubMed/NCBI
29	Cox J, Malik M, Britten J, Lewis T and Catherino WH: Ulipristal acetate and extracellular matrix production in human leiomyomas in vivo: A laboratory analysis of a randomized placebo controlled trial. Reprod Sci. 25:198–206. 2018. View Article : Google Scholar :
30	Graubner FR, Boos A, Aslan S, Kucukaslan I and Kowalewski MP: Uterine and placental distribution of selected extracellular matrix (ECM) components in the dog. Reproduction. 155:403–421. 2018. View Article : Google Scholar : PubMed/NCBI
31	Szostek-Mioduchowska AZ, Lukasik K, Skarzynski DJ and Okuda K: Effect of transforming growth factor -β1 on alpha-smooth muscle actin and collagen expression in equine endometrial fibroblasts. Theriogenology. 124:9–17. 2019. View Article : Google Scholar
32	Hao G, Zhai J, Jiang H, Zhang Y, Wu M, Qiu Y, Fan C, Yu L, Bai S, Sun L and Yang Z: Acetylshikonin induces apoptosis of human leukemia cell line K562 by inducing S phase cell cycle arrest, modulating ROS accumulation, depleting Bcr-Abl and blocking NF-κB signaling. Biomed Pharmacother. 122:1096772019. View Article : Google Scholar
33	Zhang C, Liang R, Gan X, Yang X, Chen L and Jian J: MicroRNA-384-5p/Beclin-1 as potential indicators for epigallocatechin gallate against cardiomyocytes ischemia reperfusion injury by inhibiting autophagy via PI3K/Akt pathway. Drug Des Devel Ther. 13:3607–3623. 2019. View Article : Google Scholar : PubMed/NCBI
34	Lv CL, Zhang T, Yan TZ, Yi GK and Gao K: MicroRNA-448 inhibits the regeneration of spinal cord injury through PI3K/AKT/Bcl-2 axis. Eur Rev Med Pharmacol Sci. 23:2719–2726. 2019.PubMed/NCBI
35	Wang S, Gao B, Yang H, Liu X, Wu X and Wang W: MicroRNA-432 is downregulated in cervical cancer and directly targets FN1 to inhibit cell proliferation and invasion. Oncol Lett. 18:1475–1482. 2019.PubMed/NCBI