体外受精-胚胎移植对早期胎盘黏着斑激酶信号通路的影响
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摘要
目的:研究辅助生殖中体外受精-胚胎移植(in vitro fertilization and embryo transfer,IVF-ET)技术对早期胎盘滋养层细胞黏着斑激酶(focal adhension kinase,FAK)信号通路基因表达的影响,探讨IVF-ET技术对早期胎盘发育和功能的影响。方法:收集IVF-ET来源的于7～8周经超声引导下减胎获得的胎盘绒毛组织作为研究组,对照组采用自然妊娠双胎7～8周人工流产术中获得的胎盘绒毛组织。利用美国Affymetrix HG-U133 Plus 2. 0基因芯片对两组胎盘绒毛组织进行芯片杂交分析,实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,qRT-PCR)验证其中8个差异表达基因,选取差异表达基因进行无监督聚类分析和生物信息学分析。结果:获得4例IVF-ET减胎绒毛组织和4例自然妊娠人工流产绒毛组织进行基因芯片检测。与自然妊娠组相比,IVF-ET组FAK信号通路中有32个基因差异表达,差异表达倍数≥2,其中12个基因上调,20个基因下调。经qRT-PCR验证,IVF-ET与自然妊娠早期胎盘绒毛中的8个FAK信号通路基因表达确实存在差异,与基因芯片检测结果一致。FAK信号通路基因定位显示,IVF-ET来源胎盘绒毛组织FAK信号通路上游基因表达受到影响,胎盘滋养层细胞通过基因表达代偿维持FAK信号通路功能基本正常。结论:IVF-ET来源和自然妊娠来源的早期胎盘FAK信号通路存在基因表达差异,差异表达基因涉及多种FAK信号通路关键功能,影响IVF-ET胎盘绒毛早期发育和功能,同时胎盘滋养层细胞通过改变相关基因表达来代偿IVF-ET技术本身的干扰,以维持FAK信号通路正常功能,满足胎盘绒毛和胎儿发育需要。
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 polymerase chain reaction(qRT-PCR),and unsupervised clustering analysis and functional bioinformatics analysis 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.
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 polymerase chain reaction(qRT-PCR),and unsupervised clustering analysis and functional bioinformatics analysis 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.
引文
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[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.
[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.
[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.
[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.
[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.
[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.
[7] Mohammed BT,Sontakke SD,Ioannidis J,et al. The adequate corpus luteum:miR-96 promotes luteal cell survival and progesterone production[J]. J Clin Endocrinol Metab,2017,102(7):2188-2198.
[8] Herzog EM,Eggink AJ,Willemsen SP,et al. Early-and lateonset preeclampsia and the tissue-specific epigenome of the placenta and newborn[J]. Placenta,2017,58:122-132.
[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.
[10] Labarrere CA,Di Carlo HL,Bammerlin E,et al. Failure of physiologic 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.
[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.
[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.
[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.
[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.
[16] He Y,Bai J,Liu P,et al. miR-494 protects pancreaticβ-cell function by targeting PTEN in gestational diabetes mellitus[J].EXCLI J,2017,16:1297-1307.
[17] Mathew SA,Chandravanshi B,Bhonde R. Hypoxia primed placental mesenchymal stem cells for wound healing[J]. Life Sci,2017,182:85-92.
[18] Guo M,Zhao X,Yuan X,et al. Elevated microRNA-34a contributes to trophoblast cell apoptosis in preeclampsia by targeting BCL-2[J]. J Hum Hypertens,2017,31(12):815-820.
[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.
[20] Choux C,Binquet C,Carmignac V,et al. The epigenetic control of transposable elements and imprinted genes in newborns is affected by the mode of conception:ART versus spontaneous conception without underlying infertility[J]. Hum Reprod,2018,33(2):331-340.
[21] Nakamura K,Kusama K,Bai R,et al. Increase in complement i C3b is associated with anti-inflammatory cytokine expression during late pregnancy in mice[J]. PLo S One,2017,12(5):8442-8456.
[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.
[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.
[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.