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Journal of Peking University(Health Sciences) ›› 2019, Vol. 51 ›› Issue (1): 151-158. doi: 10.19723/j.issn.1671-167X.2019.01.026

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In vitro fertilization-embryo transfer affects focal adhension kinase signaling pathway in early placenta

Liang ZHAO1,△(),Li-fang SUN1,Xiu-li ZHENG1,Jing-fang LIU1,Rong ZHENG1,Ying WANG2,Rui YANG2,Lei ZHANG3,Li YU4,Han ZHANG1   

  1. 1. Department of Obstetrics and Gynecology, Beijing Jishuitan Hospital, Beijing 100035, China
    2. Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, China
    3. Department of Obstetrics and Gynecology, Beijing Tsinghua Changgung Hospital, Beijing 102218, China
    4. Department of Obstetrics and Gynecology, Peking University First Hospital, Beijing 100034, China
  • Received:2018-04-23 Online:2019-02-18 Published:2019-02-26
  • Contact: Liang ZHAO E-mail:zhaol1972@aliyun.com
  • Supported by:
    Supported by the National Natural Science Foundation of China(81070493)

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Abstract:

Objective: To study the effects of in vitro fertilization-embryo transfer (IVF-ET) technique on gene expression of focal adhension kinase (FAK) signaling pathway in early placental trophoblast cells, and to explore the effects of IVF-ET technology on the development and function of early placenta. Methods: We collected 7-8 weeks of gestation placenta tissue as a study group by ultrasound guided reduction of fetal from double embryo transfer under IVF-ET technology. In the control group, placenta tissues were obtained from the spontaneous abortion of natural pregnancy twin 7-8 weeks. Microarray hybridization analysis was performed on the placenta tissue of the two groups using the Affymetrix HG-U133 Plus 2.0 gene chip. Eight differentially expressed genes were identified by real-time quantitative polyme-rase chain reaction (qRT-PCR), and unsupervised clustering analysis and functional bioinformatics analy-sis were performed for the differentially expressed genes. Results: Twenty-eight cases of IVF-ET reduced fetal villi and 8 cases of spontaneous abortion villi were collected. A total of 8 placental villi were detected by the gene chip. Compared with the natural pregnancy control group, 32 differentially expressed genes in the placental FAK signaling pathway were expressed in IVF-ET. The differential expression was greater than or equal to 2 times, of which 12 genes were up-regulated and 20 were down-regulated. The qRT-PCR showed that the expression of the 8 genes in FAK signaling pathways of IVF-ET was significantly different from that in the placenta of natural pregnancy, which was consistent with the result of the gene chip detection. The FAK signal pathway gene localization showed that the FAK gene was mainly located in the upstream of the signal pathway in the placenta of IVF-ET. The placental trophoblast cells maintained the FAK signaling pathway function through gene expression compensation. Conclusion: There are gene expression differences in the FAK signaling pathway between the IVF-ET derived early placenta and the natural pregnancy placenta. The differentially expressed genes are involved in many key functions of the FAK signaling pathway and affect the early development and function of the IVF-ET placenta, while the placental trophoblast cells change gene expression for interference to compensate for IVF-ET technology itself, maintain normal function of the FAK signaling pathway, and satisfy the need for placental and fetal development.

Key words: Fertilization in vitro, Embryo transfer, Trophoblasts, Focal adhension kinase, Gene expression

CLC Number: 

  • R321-33

Table 1

General information on gene chip detection cases"

Case number Age/years Number of pregnancies Number of births Pregnancy time
Control 1 34 3 0 7 weeks+1
Control 2 34 3 0 7 weeks+5
Control 3 31 1 0 7 weeks+2
Control 4 33 2 0 7 weeks+6
IVF-ET 1 34 3 1 7 weeks+2
IVF-ET 2 30 2 0 7 weeks+6
IVF-ET 3 33 3 1 7 weeks+1
IVF-ET 4 34 3 1 7 weeks+3

Figure 1

Gene chip detection of FAK signaling pathway differential gene Heatmap analysis The expression of FAK signaling pathway genes in the placenta tissue of IVF-ET group and control group, the red represents gene up-regulated and the green represents gene expression down-regulated. VTN, vitronectin; ECM, extracellular matrix protein; ITGA, integrin, alpha; PDGFRA, platelet-derived growth factor receptor, alpha polypeptide; JUN, Jun proto-oncogene; PDGFC, platelet derived growth factor C; PDGFD, platelet derived growth factor D; CCND1, cyclin D1; FYN, FYN oncogene related to SRC; SHC, SHC (Src homology 2 domain containing) transforming protein 2; GSK3B, glycogen synthase kinase 3 beta; PAK, P21 protein (Cdc42/Rac)-activated kinase; BRAF, V-raf murine sarcoma viral oncogene homolog B1; ITGB3, integrin, beta 3; CAV2, caveolin 2; ERBB2, V-erb-b2 erythroblastic leukemia viral oncogene homolog 2; ITGB6, integrin, beta 6; FN1, fibronectin 1; CAV1, caveolin 1; HGF, hepatocyte growth factor; VAV3, Vav 3 guanine nucleotide exchange factor; PTEN, phosphate and tension homology deleted on chromsome ten; ITGA5, integrin, alpha 5; ITGB4, integrin, beta 4; FLT1, Fms-related tyrosine kinase 1; PAK6, P21 protein (Cdc42/Rac)-activated kinase 6; PIK3CB, phosphoinositide-3-kinase, catalytic, beta polypeptide; PGF, placental growth factor; MAPK9, mitogen-activated protein kinase 9; LAMA2, laminin, alpha 2; BCL2, B-cell lymphoma 2; EGFR, epidermal growth factor receptor."

Figure 2

Double log scatter plot of gene chip detection FAK signaling pathway differential gene The double-log scatter plot showed that the expression of FAK signaling pathway genes were different by gene chip in the IVF-ET group and the control group. Red pots represent up-regulation of gene expression; Green pots represent down-regulation of gene expression."

Figure 3

Unsupervised cluster analysis of differential genes in FAK signaling pathway by gene chip Unsupervised cluster analysis showed FAK signaling pathway differential gene expression between placental villus tissues, red for IVF-ET and green for control."

Figure 4

Effect of IVF-ET on gene expression of FAK signaling pathway in placental villi The changes in gene expression in the FAK signaling pathway of placental villus in the IVF-ET technique and the position and mutual regulation of the genes are shown.Red represents up-regulation of gene expression and green represents down-regulation of gene expression.Abbreviations as in Figure 1."

Figure 5

qRT-PCR comparison of differential expression of genes in FAK signaling pathway in IVF-ET and control placenta qRT-PCR results showing differential expression of eight FAK signaling pathway genes, including JUN, GSK3B, PDGFRA, ITGA, MAPK9, EGFR, FN1 and BCL2, showing up-regulated genes (red columns), down-regulated genes (green columns) and control groups (blue column). Abbreviations as in Figure 1. All data were x ? ±s. *P<0.05, **P<0.01. "

[1] Ata B, Mumusoglu S, Aslan K , et al. Which is worse? A compa-rison of ART outcome between women with primary or recurrent endometriomas[J]. Hum Reprod, 2017,32(7):1539-1549.
doi: 10.1093/humrep/dex099 pmid: 28498960
[2] Shapiro AJ, Darmon SK, Barad DH , et al. Effect of race and ethnicity on utilization and outcomes of assisted reproductive technology in the USA[J]. Reprod Biol Endocrinol, 2017,15(1):44-60.
doi: 10.1186/s12958-017-0262-5 pmid: 28595591
[3] Luo WW, Zhao WW, Lu JJ , et al. Cucurbitacin B suppresses metastasis mediated by reactive oxygen species (ROS) via focal adhesion kinase (FAK) in breast cancer MDA-MB-231 cells[J]. Chin J Nat Med, 2018,16(1):10-19.
doi: 10.1016/S1875-5364(18)30025-6 pmid: 29425586
[4] Zhang Y, Cui Y, Zhou Z , et al. Altered global gene expressions of human placentae subjected to assisted reproductive technology treatments[J]. Placenta, 2010,31(4):251-258.
doi: 10.1016/j.placenta.2010.01.005 pmid: 20116094
[5] Sakian S, Louie K, Wong EC , et al. Altered gene expression of H19 and IGF2 in placentas from ART pregnancies[J]. Placenta, 2015,36(10):1100-1105.
doi: 10.1016/j.placenta.2015.08.008 pmid: 26386650
[6] Zhan Q, Qi X, Wang N , et al. Altered methylations of H19, Snrpn, Mest and Peg3 are reversible by developmental reprogramming in kidney tissue of ICSI-derived mice[J]. Sci Rep, 2017,7(1):11936-11944.
doi: 10.1038/s41598-017-11778-w pmid: 28931827
[7] Mohammed BT, Sontakke SD, Ioannidis J , et al. The adequate corpus luteum: miR-96 promotes luteal cell survival and progeste-rone production[J]. J Clin Endocrinol Metab, 2017,102(7):2188-2198.
doi: 10.1210/jc.2017-00259 pmid: 28368475
[8] Herzog EM, Eggink AJ, Willemsen SP , et al. Early- and late-onset preeclampsia and the tissue-specific epigenome of the placenta and newborn[J]. Placenta, 2017,58:122-132.
doi: 10.1016/j.preghy.2017.07.075 pmid: 28962690
[9] 赵亮, 孙丽芳, 郑秀丽 , 等. 植入前囊胚滋养外胚层基因空间表达[J]. 北京大学学报(医学版), 2017,49(6):965-973.
doi: 10.3969/j.issn.1671-167X.2017.06.006
[10] Labarrere CA, DiCarlo HL, Bammerlin E , et al. Failure of phy-siologic transformation of spiral arteries, endothelial and trophoblast cell activation, and acute atherosis in the basal plate of the placenta[J]. Am J Obstet Gynecol, 2017,216(3):287-297.
[11] Song GY, Na Q, Wang D , et al. Microarray expression profile of lncRNAs and mRNAs in the placenta of non-diabetic macrosomia[J]. J Dev Orig Health Dis, 2018,9(2):191-197.
doi: 10.1017/S2040174417000927 pmid: 29141697
[12] Xin L, Xu B, Ma L , et al. Proteomics study reveals that the dysregulation of focal adhesion and ribosome contribute to early pregnancy loss[J]. Proteomics Clin Appl, 2016,10(5):554-563.
doi: 10.1002/prca.201500136 pmid: 26947931
[13] Hilal T, Fauble V, Ketterling RP , et al. Myeloid neoplasm with eosinophilia associated with isolated extramedullary FIP1L1/PDGFRA rearrangement[J]. Cancer Genet, 2018,220(1):13-18.
doi: 10.1016/j.cancergen.2017.10.004 pmid: 29310833
[14] Zhang A, Lakshmanan J, Motameni A , et al. MicroRNA-203 suppresses proliferation in liver cancer associated with PIK3CA, p38MAPK, c-Jun, and GSK3 signaling[J]. Mol Cell Biochem, 2018,441(1-2):89-98.
doi: 10.1007/s11010-017-3176-9 pmid: 28887744
[15] Wu L, Hafiz MZ, Guan Y , et al. 17β-estradiol suppresses carboxylesterases by activating c-Jun/AP-1 pathway in primary human and mouse hepatocytes[J]. Eur J Pharmacol, 2018,15(8):98-107.
doi: 10.1016/j.ejphar.2017.11.036 pmid: 29175444
[16] He Y, Bai J, Liu P , et al. miR-494 protects pancreatic β-cell function by targeting PTEN in gestational diabetes mellitus[J]. EXCLI J, 2017,16:1297-1307.
doi: 10.17179/excli2017-491 pmid: 29333131
[17] Mathew SA, Chandravanshi B, Bhonde R . Hypoxia primed placental mesenchymal stem cells for wound healing[J]. Life Sci, 2017,182:85-92.
doi: 10.1016/j.lfs.2017.06.016 pmid: 28625360
[18] Guo M, Zhao X, Yuan X , et al. Elevated microRNA-34a contri-butes to trophoblast cell apoptosis in preeclampsia by targeting BCL-2[J]. J Hum Hypertens, 2017,31(12):815-820.
doi: 10.1038/jhh.2017.65 pmid: 29022890
[19] Jiang C, Xu M, Kuang X , et al. Treponema pallidum flagellins stimulate MMP-9 and MMP-13 expression via TLR5 and MAPK/NF-κB signaling pathways in human epidermal keratinocytes[J]. Exp Cell Res, 2017,361(1):46-55.
doi: 10.1016/j.yexcr.2017.09.040 pmid: 28982539
[20] Choux C, Binquet C, Carmignac V , et al. The epigenetic control of transposable elements and imprinted genes in newborns is affec-ted by the mode of conception: ART versus spontaneous conception without underlying infertility[J]. Hum Reprod, 2018,33(2):331-340.
doi: 10.1093/humrep/dex366 pmid: 29237055
[21] Nakamura K, Kusama K, Bai R , et al. Increase in complement iC3b is associated with anti-inflammatory cytokine expression during late pregnancy in mice[J]. PLoS One, 2017,12(5):8442-8456.
doi: 10.1371/journal.pone.0178442 pmid: 28542608
[22] Song X, Rui C, Meng L , et al. Long non-coding RNA RPAIN regulates the invasion and apoptosis of trophoblast cell lines via complement protein C1q[J]. Oncotarget, 2017,8(5):7637-7646.
doi: 10.18632/oncotarget.13826 pmid: 28032589
[23] Weinerman R, Ord T, Bartolomei MS , et al. The superovulated environment, independent of embryo vitrification, results in low birthweight in a mouse model[J]. Biol Reprod, 2017,97(1):133-142.
doi: 10.1093/biolre/iox067 pmid: 28859279
[24] Martin E, Smeester L, Bommarito PA , et al. Sexual epigenetic dimorphism in the human placenta: implications for susceptibility during the prenatal period[J]. Epigenomics, 2017,9(3):267-278.
doi: 10.2217/epi-2016-0132 pmid: 28234023
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