Skip to main content
NCBI home page
International Journal of Clinical and Experimental Pathology logoLink to International Journal of Clinical and Experimental Pathology
. 2018 Jan 1;11(1):110–119.

Upregulated miR-222 targets BCL2L11 and promotes apoptosis of mesenchymal stem cells in preeclampsia patients in response to severe hypoxia

Hong-Mei Qu 1,*, Li-Ping Qu 1,*, Xian-Zhen Pan 1,*, Lin-Song Mu 2
PMCID: PMC6957949  PMID: 31938092

Abstract

Abnormal maternal trophoblast invasion is a common finding in preeclampsia pregnancy. A hypoxic environment develops in the placenta after the 10th week of pregnancy and exerts a major influence over trophoblast activity. In the present study, we investigated expression of miR-222 and apoptosis-related BCL2L11 in preeclampsia placenta and in primary placenta mesenchymal stem cells (MSCs) under hypoxia. The results demonstrate that miR-222 is upregulated in the placenta of preeclampsia patients, along with the downregulation of BCL2L11 in both mRNA and protein levels. In vitro results demonstrate that miR-222 is upregulated either in preeclampsia placenta tissues or in the MSCs under hypoxia. Western blotting showed downregulation of BCL2L11 in the trophoblasts under hypoxia, along with an increased MSC apoptosis. miR-222 was also confirmed to downregulate BCL2L11 expression via targeting the 3’ untranslated region (UTR) of the BCL2L11 gene. miR-222 inhibitor transfection markedly ameliorated expression and transcriptional activity of BCL2L11. Altogether, the present study found that upregulation of miR-222 promotes apoptosis of mesenchymal stem cells in preeclampsia patients in response to hypoxia, via targeting BCL2L11. This suggests that a key regulatory role of miR-222 is in preeclampsia progression.

Keywords: Apoptosis, BCL2L11, hypoxia, miR-222, preeclampsia

Introduction

Preeclampsia is an abnormal pregnancy-related syndrome with proteinuria and hypertension following 20-weeks of gestation [1]. Preeclampsia affects 3-5% of pregnancies worldwide and 9.4-10.4% of Chinese pregnancies every year, and is a leading risk factor for maternal, fetal, and neonatal mortality [2-4]. Preeclampsia is currently recognized to associate with inadequate trophoblast invasion and placentation [3], with decreased circulation in the early placenta, and with continuous placental ischemia after 20 weeks of gestation [5,6]. The sustained ischemia/hypoxia promotes oxidative stress, and thus induces production of excessive carbon monoxide, nitric oxide, and androgen in the local microcirculation [7]. Thus, increased apoptosis of trophoblast cells in the placenta, which are driven by the oxidative stress, can decrease trophoblast invasion and may be the major pathological cause of preeclampsia [8,9].

Mesenchymal stem cells (MSCs) represent a group of cells with multi-differentiation potential [10]. MSCs are mainly from bone marrow and human umbilical cord and have high regenerative capacities [11,12]. MSCs are capable of differentiating into a wide variety of cell types [13] and display key roles in tissue regeneration and repair [14]. Interestingly, human umbilical cord MSCs from preeclampsia patients show differentiation, indicating clinical potential for the preeclampsia treatment [15]. Recent findings in mouse models indicated that MSCs have a significant influence on the course of preeclampsia [16].

microRNAs (miRNAs) are a group of short non-coding single-stranded RNA molecules, which inhibit mRNA translation or promote mRNA degradation through specific binding mainly to the 3’-untranslated region (UTR) of targeted genes [17]. miRNAs have been recognized to be involved in the pathogenesis of preeclampsia. Abnormal miRNA profiles have been identified in placental tissues [18], placental trophoblasts [19], and placental MSCs [20], plasma/whole blood samples [21]. Accumulating endothelial-related miRNAs have been reported to be potentially important in hypoxia-promoted endothelial damage in preeclampsia [20,21]. In particular, miRNAs, such as miR-16, miR-181a, and miR-494 are abnormally expressed in placental MSCs and inhibit the angiogenesis potential at the maternal-fetal interface in preeclampsia [16,22-24]. Therefore, miRNAs may act as an important regulators involved in the pathogenesis of preeclampsia.

In this study, we analyzed expression of miR-222 and BCL2L11 (B-cell lymphoma 2) in placental tissues, and in MSCs from patients with preeclampsia. We explored miR-222 or BCL2L11 expression in MSCs under normoxia or hypoxia. Finally we investigated the role of miR-222 in hypoxia-induced apoptosis and its effect on expression of BCL2L11 in hypoxia-treated MSCs. Our findings contribute to a greater understanding of miRNA function in preeclampsia MSCs.

Materials and methods

Placenta specimens

17 pregnant women with preeclampsia and 17 normal pregnant subjects were included in this study. All included subjects were registered at the Department of Obstetrics and Gynecology, the Affiliated Yantai Yuhuangding Hospital of Qingdao University and formally signed consent at the time of study enrollment. The present study was approved by the Ethics Committee of our hospital. Detailed clinic-pathological information was reported previously (https://doi.org/10.5114/aoms.2016.63261).

Isolation and culture of MSCs

Placentas from two groups of donor mothers were obtained immediately post-delivery. For BCL2L11 immuno-staining, placenta sections were fixed with 10% formalin and were embedded with paraffin. Decidua basalis was dissected from the maternal-facing surface of placenta, and were immediately set in ice-cold phosphate buffered saline (PBS) for the MSC isolation. The MSC isolation was performed according to a reported protocol [25]. In brief, decidua basalis was mechanically minced, and enzymatically digested with 0.1% collagenase IV (Thermo Fisher Scientific, Inc., Waltman, MA, USA) to produce the cell suspension. Following a series of centrifugation steps (with or without Ficoll) and washes with PBS, the cell pellet was suspended in the complete low-glucose Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, Rockville, MD, USA), supplemented with 10% fetal bovine serum (FBS) (Ameresco, Framingham, MA, USA).

MSCs were seeded and cultured in 24-well plates with complete medium. 6-10 days post culture, the fibroblast-like cells adhered in the well during primary passage and were harvested using 0.25% trypsinase (Gibco-BRL, Hangzhou, China), and then seeded in 24-well plates at the density of 5×104 cells/cm2 for secondary passage. The non-adherent cells were harvested and transferred into another 24-well plate by updating 1/2 medium. Cells were cultured at 37°C under 5% CO2 and 20% O2 routinely. Hypoxia treatment in vitro was simulated by the chemical hypoxia agent CoCl2 (100 mmol/L).

Quantitative real-time polymerase chain reaction for BCL2L11 mRNA and miR-222

PureLink® RNA Mini Kit (Invitrogen, Carlsbad, CA, USA) and mirVanaTM miRNA Isolation Kit (Ambion, Austin, TX, USA) were utilized for the mRNA and miRNA isolation respectively. Each sample of mRNA or miRNA was supplemented with ribonuclease inhibitor (Promega, Madison, WI, USA) and was was quantified for BCL2L11 mRNA level or for miR-222 level by the quantitative real-time PCR (qRT-PCR) method with Takara One Step RT-PCT kit (Takara, Tokyo, Japan) (for mRNA) or with mirVanaTM qRT-PCR miRNA Detection Kit (Thermo Scientific, Rockford, IL, USA) (for miRNA). Relative levels of BCL2L11 mRNA or miR-222 were calculated using the ΔΔCt method using β-actin or U6 as internal control gene [26].

Immuno-staining for BCL2L11 in placenta tissues

Paraffinized placenta tissue slices were firstly deparaffinized, and subjected to antigen resurrection according to a general standard protocol. Polyclonal rabbit anti-BCL2L11 (Cell Signaling Technology Inc., Danvers, MA, USA) and HRP-coupled goat anti-rabbit IgG antibodies (Abcam, Cambridge, UK) were utilized for the BCL2L11 immuno-staining in the placental tissue. 3-amino-9-ethylcarbazole (AEC) (Sigma-Aldrich, St. Louis, MO, USA) was utilized as an HRP substrate for staining and the immune-staining was semi-quantified by counting AEC-positive cells.

Western blot analysis

Protein samples from placental tissue or MSCs were isolated with NE-PER Nuclear and Cytoplasmic Extraction Reagents Kit (Pierce, Rockford, IL, USA) under the guidance of kit’s manual. For the Western blot of BCL2L11, each sample was separated by electrophoresis (12% SDS-PAGE gel), transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), blocked with 2% BSA (Sigma-Aldrich, St. Louis, MO, USA) and then the membrane was incubated successively with rabbit polyclonal antibody against human BCL2L11 or β-actin respectively at room temperature for 2 hours. HRP-conjugated secondary antibody against rabbit IgG at 4°C was added for another one hour, and finally electrochemiluminescence (ECL) (Amersham, Uppsala, Sweden) was performed at room temperature for 25 minutes. The levels of BCL2L11 are presented as a percent gray value normalized to β-actin.

Luciferase reporting assay

The 3’UTR of BCL2L11 and the CMV (Cytomegalovirus) promoter were cloned into the pGL3-luciferase basic vector (Promega, Madison, WI, USA). Sequences of primers and cloning strategy are available on request. For the luciferase assays, 50 nM hsa-miR-222 miRNA precursor (Thermo Scientific, Rockford, IL, USA) or scrambled Pre-miRTM miRNA Precursor Negative Control (Thermo Scientific, Rockford, IL, USA) were transfected with siPORTTM NeoFXTM Transfection Agent (Thermo Scientific, Rockford, IL, USA) into the cultured 80%-confluent MSCs, 2 hours later, MSCs were then transfected with the pGL3-luciferase reporter vector and the Renilla control vector (Promega, Madison, WI, USA) by lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). 24 h post transfection, the Dual Luciferase reporter assay kit (Promega, Madison, WI, USA) was utilized to examine luciferase activity.

Apoptosis assay

Annexin V/Propidium Iodide double staining was performed to determine the placental MSC apoptosis using an Annexin V-FITC Apoptosis Detection Kit (Abcam, Cambridge, UK). In brief, 1×104 MSCs per ml aliquot were suspended in PBS, and 5 μl annexin V-FITC and 10 μl PI were added, followed by incubation for 10 min at 4°C in the dark. Finally, the stained cells were visualized with a live cell imaging system (Olympus LCS SYSTEM, Tokyo, Japan). Early apoptotic MSCs (Annexin V+/PI-) and late apoptotic MSCs (Annexin V+/PI+) were expressed as apoptotic MSCs to total MSCs.

MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay

MTT cell viability assay kit (Biotium Inc, Beijing, China) was utilized to examine the MSC viability. MSCs were seeded in 96-well plates to approximately 85% confluence and then cells were incubated at 37°C under hypoxia or normoxia for 6 or 12 hours. 10 μL MTT was added to each well and was incubated at 37°C for another 2 hours. Finally, 100 μl DMSO was added into each well to dissolve the formazan, and the absorbance was measured with a test wavelength of 570 nm and a reference wavelength of 630 nm.

Statistical analysis

Quantitative data were presented as means ± SEM. A nonparametric Mann-Whitney test or ANOVA StatView was used for data analysis on GraphPad Prism 5 (GraphPad Software, La Jolla, CA, USA). Statistical significance was considered when P<0.05 or less.

Results

Upregulation of miR-222 in association with BCL2L11 downregulation in preeclampsia placentas specimens

Significant increase of miR-222 has been found in maternal plasma at the time of severe preeclampsia [27]. However, it is not clear whether its expression in preeclamptic placental tissues is of functional significance. To investigate the miR-222 level in preeclampsia placentas, we quantified miR-222 (relative to U6) in 17 preeclampsia subjects. As indicated in Figure 1A, the relative miR-222 level was upregulated in the preeclampsia group (P = 0.0022). Various genes, including BCL2L11, are targeted by miR-222 [28,29]. Therefore, BCL2L11 expression in the two groups of samples was examined and it is shown in Figure 1B that BCL2L11 mRNA level was significantly downregulated in the preeclampsia group (P = 0.0014). Moreover, there was a negative association of BCL2L11 downregulation with the miR-222 upregulation (P = 0.0064, R2 = 0.4010, Figure 1C).

Figure 1.

Figure 1

Open in a new tab

Quantification of miR-222 and BCL2L11 mRNA in preeclampsia placental specimens. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to measure the relative miR-222 (A) (to U6) and the relative BCL2L11 mRNA (B) (to β-actin) in the placental specimens from preeclampsia (N = 17) or normal (N = 17) subjects. (C) Correlation of miR-222 level with BCL2L11 mRNA in the placental specimens from preeclampsia subjects. Statistical significance with a P value <0.05 or less.

Immunohistochemical analysis was also performed to verify the BCL2L11 downregulation in the preeclampsia group. In contrast to staining for BCL2L11 in the placentas tissues of the control group (Figure 2A), there were fewer stained cells in the preeclampsia placentas tissues (Figure 2B), which was confirmed by quantitative analysis (P<0.001, Figure 2C). Thus, we found that upregulation of miR-222 in association with BCL2L11 downregulation was prominent in preeclampsia placentas specimens.

Figure 2.

Figure 2

Open in a new tab

Immunohistochemical staining for BCL2L11 in preeclampsia placental specimens. BCL2L11 staining by immunofluorescence on the placenta tissues from normal (n = 17) (A) or from preeclampsia (n = 17) (B) subjects. The BCL2L11 positive staining is indicated with white arrows. (C) BCL2L11-positive cell number in preeclampsia and normal groups. Statistical significance with a P value <0.05 or less.

Hypoxia upregulates miR-222 and downregulates BCL2L11 in placental mesenchymal stem cells (MSCs)

The role of hypoxia in promoting preeclampsia has been identified previously [30,31]. We were thus interested in whether hypoxia regulates both miR-222 and BCL2L11 in human placental cells. Human placenta mesenchymal stem cells (MSCs) were cultured under hypoxia and normoxia and then expression of both molecules was examined. Figure 3A demonstrates a higher level of miR-222 in hypoxia-treated MSCs, either at 6 or 12 hours post treatment (P<0.05 or P<0.01). On the other hand, BCL2L11 mRNA level (relative to β-actin) was downregulated in the hypoxia group (P<0.05 or P<0.01 for 6 or 12 hours, Figure 3B). Such downregulation was also confirmed at the protein level by Western blotting at 12 or 24 hour post hypoxia (P<0.05 or P<0.01, Figure 2C and 2D).

Figure 3.

Figure 3

Open in a new tab

Expression of miR-222 and BCL2L11 in placental MSCs under hypoxia. Approximately 85%-confluent MSCs were cultured under normoxia or hypoxia for another 6, 12, or 24 hours, then qRT-PCR was performed to measure the miR-222 (A) and BCL2L11 mRNA (B) levels in MSCs under normoxia or hypoxia for 6 or 12 hours. (C and D) Western blotting (C) and quantification (D) of BCL2L11 in the normoxia- or hypoxia-treated MSCs under normoxia or hypoxia for 12 or 24 hours. Triple independent experiments were performed. *P<0.05 or **P<0.01.

miR-222 downregulates BCL2L11 via targeting the 3’UTR of BCL2L11 gene in MSCs

BCL2L11 is identified as one target for miR-222 [28,29]. We then investigated such targeting regulation by miR-222 on the BCL2L11 gene in placental MSCs. Manipulated upregulation of miR-222 with miR-222 mimics (P<0.0001, Figure 4A) markedly downregulated BCL2L11 mRNA level in MSCs with a transfection concentration of 20 or 40 nM, with scrambled RNA as a control (Ctrl miRNA) (P<0.05 or P<0.01, Figure 4B). The BCL2L11 protein level was also significantly downregulated by transfection of miR-222 mimics at a concentration of 20 or 40 nM (P<0.05 or P<0.01, Figure 4C).

Figure 4.

Figure 4

Open in a new tab

Downregulation by miR-222 on BCL2L11 in MSCs in vitro. Approximately 85%-confluent MSCs were transfected with 20 or 40 nM miR-222 mimics (to U6) or scrambled RNA (Ctrl miRNA) for 12 or 24 hours, and the relative miR-222 level (to U6) (A) and the relative mRNA level of BCL2L11 (to β-actin) (B) were quantified. (C and D) Western blot analysis (C) of and the quantification (D) of BCL2L11 protein in the miR-222 mimics- or the scrambled RNA-transfected (for 24 hours) MSCs. Triple independent experiments were performed. *P<0.05, **P<0.01 or ****P<0.0001.

In addition, we also constructed a luciferase-based reporter for the paired sequence (Figure 5A) to miR-222 in the 3’UTR of the BCL2L11 gene (Figure 5B). A control reporter was also constructed with a mutant 3’UTR sequence (unpaired to miR-222) of the BCL2L11 gene (Figure 5B). The luciferase reporter assay confirmed that transfection with 20 or 40 nM miR-222 mimics markedly reduced luciferase activity in MSCs, compared to control miRNA (P<0.01 or P<0.001, Figure 5C). However, such differential regulation was not found between miR-222 and control miRNA with the mutant reporter (Figure 5D).

Figure 5.

Figure 5

Open in a new tab

Regulation by miR-222 on BCL2L11 expression via targeting 3’UTR of BCL2L11 in MSCs with a luciferase reporter. (A and B) Sequence analysis of Homo sapiens miR-222 and the 3’UTR of Homo sapiens BCL2L11 (A) and a sketch (B) of the luciferase reporter with wild-type and mutant 3’UTR of BCL2L11 inserted behind the cytomegalovirus promoter. (B and C) Relative luciferase in MSCs, which were transfected with the reporter with wild-type (C) or mutant (D) 3’UTR of BCL2L11. Triple independent experiments were performed. **P<0.01, ****P<0.0001, ns: no significance.

miR-222 regulates sensitivity of MSCs to hypoxia

Cellular viability and apoptosis were examined respectively with MTT assay and flow cytometry analysis. Hypoxia treatment significantly reduced cellular viability of MSCs for either 6 or 12 hours (P<0.05 or P<0.01, Figure 6A). Moreover, a significantly higher apoptosis was also induced in MSCs by hypoxia for 12 or 24 hours (P<0.05 or P<0.01, Figure 6B). To evaluate the role of hypoxiapromoted miR-222 in the hypoxia-mediated cellular viability reduction and apoptosis induction, we knocked down miR-222 in the hypoxia-treated MSCs, and then re-examined cell viability and apoptosis. Figure 6C demonstrates that transfection with 20 or 40 nM miR-222 inhibitor significantly knocked down the miR-222 level (P<0.01 respectively). Moreover, the miR-222 inhibitor-mediated miR-222 knockdown with 40 nM markedly ameliorated the hypoxia-induced loss of MSC viability (P<0.01, Figure 6D). Hypoxia-induced apoptosis was also reduced (P<0.05 for 20 nM or P<0.01 for 40 nM, Figure 6E).

Figure 6.

Figure 6

Open in a new tab

miR-222 regulates hypoxia-induced cellular viability reduction and apoptosis in MSCs. (A and B) Relative cellular viability (A) and apoptosis (B) of the MSCs, which were cultured under normoxia or hypoxia for 6, 12 or 24 hours. Cellular viability was examined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and cell apoptosis was determined by flow cytometry analysis. (C) Relative miR-222 level (to U6) in the MSCs, which were transfected with 20 or 40 nM miR-222 inhibitor (with Scramble RNA as control) for 24 hours. (D and E) Relative cellular viability (D) and apoptosis (E) of the MSCs, post the miR-222 inhibitor- or scrambled RNA-transfection (20 or 40 nM) and under hypoxia. Triple independent experiments were performed. *P<0.05, **P<0.01, ns: no significance.

Discussion

Increasing evidence indicates that there are impairments in preeclampsia [32] affecting trophoblast-mediated implantation and placentation, which are essential for the future development of embryo organisms [33,34]. In particular, apoptosis of trophoblasts is increased in preeclampsia [35-38]. Therefore, placental hypoxia developed during preeclampsia represents one of the most common causes of placental trophoblast damage via apoptosis [37,39,40]. Our previous study also showed a promoting role for hypoxia in preeclampsia [41].

Significant increases of miR-222 have been found in maternal plasma at the time of severe preeclampsia [27]. We found here that miR-222 is upregulated in preeclampsia placentas specimens. Therefore, we speculate that miR-222 might play a role in preeclampsia. We also found that BCL2L11 was significantly downregulated in both mRNA and protein levels in preeclampsia placentas specimens. BCL2L11 is targeted by miR-222 [28,29] and there is a negative association between the two deregulated molecules. This data implies that miR-222-mediated regulation might depend on targeting BCL2L11. Our in vitro results confirmed negative regulation by miR-222 on BCL2L11 via targeting the 3’UTR of BCL2L11 gene in human placental MSCs. Manipulated upregulation of miR-222 with miR-222 mimics also markedly downregulated BCL2L11 in both mRNA and protein levels in MSCs with a concentration of 20 and 40 nM. Moreover, a luciferase-based approach using the paired sequence of BCL2L11 gene indicated that miR-222 transfection markedly reduces luciferase activity in MSCs. Thus, we confirmed negative regulation by miR-222 on BCL2L11 in human placental MSCs.

Hypoxia has been identified to promote the development of preeclampsia [30,31]. Our results further demonstrate that hypoxia upregulates miR-222 and downregulates BCL2L11 in placental MSCs. A higher level of miR-222 and reduced mRNA and protein levels of BCL2L11 were found in hypoxia-treated MSCs. The mechanism of the promotion by hypoxia to preeclampsia has not been clear. Our study indicates that hypoxia markedly downregulates cellular viability and promotes apoptosis of MSCs in vitro. Furthermore, hypoxia-upregulated miR-222 regulates the sensitivity of MSCs to hypoxia. The knockdown of miR-222 with a miR-222 inhibitor in the hypoxia-treated MSCs also significantly ameliorated the hypoxia-caused cellular viability reduction and the hypoxia-induced apoptosis in MSCs.

Taken together, we included that miR-222 is upregulated in preeclampsia placenta, associating with a reduced BCL2L11. miR-222 also modulates hypoxia-induced damage to human placental MSCs by downregulating BCL2L11 and inducing apoptosis. Our study implies that miR-222/BCL2L11 might be an important signal in preeclampsia.

Acknowledgements

The present study was supported by the grant from the National Nature Science Foundation of China (No. 81601288).

Disclosure of conflict of interest

None.

References

  • 1.Phipps E, Prasanna D, Brima W, Jim B. Preeclampsia: updates in pathogenesis, definitions, and guidelines. Clin J Am Soc Nephrol. 2016;11:1102–13. doi: 10.2215/CJN.12081115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Duley L. The global impact of pre-eclampsia and eclampsia. Semin Perinatol. 2009;33:130–37. doi: 10.1053/j.semperi.2009.02.010. [DOI] [PubMed] [Google Scholar]
  • 3.Steegers EA, von Dadelszen P, Duvekot JJ, Pijnenborg R. Pre-eclampsia. Lancet. 2010;376:631–44. doi: 10.1016/S0140-6736(10)60279-6. [DOI] [PubMed] [Google Scholar]
  • 4.Ghulmiyyah L, Sibai B. Maternal mortality from preeclampsia/eclampsia. Semin Perinatol. 2012;36:56–59. doi: 10.1053/j.semperi.2011.09.011. [DOI] [PubMed] [Google Scholar]
  • 5.Red-Horse K, Zhou Y, Genbacev O, Prakobphol A, Foulk R, McMaster M, McMaster M, Fisher SJ. Trophoblast differentiation during embryo implantation and formation of the maternalfetal interface. J Clin Invest. 2004;114:744–54. doi: 10.1172/JCI22991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Genbacev O, Zhou Y, Ludlow JW, Fisher SJ. Regulation of human placental development by oxygen tension. Science. 1997;277:1669–72. doi: 10.1126/science.277.5332.1669. [DOI] [PubMed] [Google Scholar]
  • 7.Ferguson KK, McElrath TF, Chen YH, Mukherjee B, Meeker JD. Longitudinal profiling of inflammatory cytokines and C-reactive protein during uncomplicated and preterm pregnancy. Am J Reprod Immunol. 2014;72:326–36. doi: 10.1111/aji.12265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.DiFederico E, Genbacev O, Fisher SJ. Preeclampsia is associated with widespread apoptosis of placental cytotrophoblasts within the uterine wall. Am J Pathol. 1999;155:293–301. doi: 10.1016/S0002-9440(10)65123-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Ishihara N, Matsuo H, Murakoshi H, Laoag-Fernandez JB, Samoto T, Maruo T. Increased apoptosis in the syncytiotrophoblast in human term placentas complicated by either preeclampsia or intrauterine growth retardation. Am J Obstet Gynecol. 2002;186:158–66. doi: 10.1067/mob.2002.119176. [DOI] [PubMed] [Google Scholar]
  • 10.Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143–47. doi: 10.1126/science.284.5411.143. [DOI] [PubMed] [Google Scholar]
  • 11.El Omar R, Beroud J, Stoltz JF, Menu P, Velot E, Decot V. Umbilical cord mesenchymal stem cells: the new gold standard for mesenchymal stem cell-based therapies? Tissue Eng Part B Rev. 2014;20:523–44. doi: 10.1089/ten.TEB.2013.0664. [DOI] [PubMed] [Google Scholar]
  • 12.Lu LL, Liu YJ, Yang SG, Zhao QJ, Wang X, Gong W, Han ZB, Xu ZS, Lu YX, Liu D, Chen ZZ, Han ZC. Isolation and characterization of human umbilical cord mesenchymal stem cells with hematopoiesis-supportive function and other potentials. Haematologica. 2006;91:1017–26. [PubMed] [Google Scholar]
  • 13.De Schauwer C, Goossens K, Piepers S, Hoogewijs MK, Govaere JL, Smits K, Meyer E, Van Soom A, Van de Walle GR. Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources. Stem Cell Res Ther. 2014;5:6. doi: 10.1186/scrt395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Karlupia N, Manley NC, Prasad K, Schafer R, Steinberg GK. Intraarterial transplantation of human umbilical cord blood mononuclear cells is more efficacious and safer compared with umbilical cord mesenchymal stromal cells in a rodent stroke model. Stem Cell Res Ther. 2014;5:45. doi: 10.1186/scrt434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Joerger-Messerli M, Bruhlmann E, Bessire A, Wagner A, Mueller M, Surbek DV, Schoeberlein A. Preeclampsia enhances neuroglial marker expression in umbilical cord Wharton's jellyderived mesenchymal stem cells. J Matern Fetal Neonatal Med. 2015;28:464–69. doi: 10.3109/14767058.2014.921671. [DOI] [PubMed] [Google Scholar]
  • 16.Liu L, Wang Y, Fan H, Zhao X, Liu D, Hu Y, Kidd AR 3rd, Bao J, Hou Y. MicroRNA-181a regulates local immune balance by inhibiting proliferation and immunosuppressive properties of mesenchymal stem cells. Stem Cells. 2012;30:1756–70. doi: 10.1002/stem.1156. [DOI] [PubMed] [Google Scholar]
  • 17.Ouellet DL, Perron MP, Gobeil LA, Plante P, Provost P. MicroRNAs in gene regulation: when the smallest governs it all. J Biomed Biotechnol. 2006;2006:69616. doi: 10.1155/JBB/2006/69616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Weedon-Fekjaer MS, Sheng Y, Sugulle M, Johnsen GM, Herse F, Redman CW, Lyle R, Dechend R, Staff AC. Placental miR-1301 is dysregulated in early-onset preeclampsia and inversely correlated with maternal circulating leptin. Placenta. 2014;35:709–17. doi: 10.1016/j.placenta.2014.07.002. [DOI] [PubMed] [Google Scholar]
  • 19.Li Q, Pan Z, Wang X, Gao Z, Ren C, Yang W. miR-125b-1-3p inhibits trophoblast cell invasion by targeting sphingosine-1-phosphate receptor 1 in preeclampsia. Biochem Biophys Res Commun. 2014;453:57–63. doi: 10.1016/j.bbrc.2014.09.059. [DOI] [PubMed] [Google Scholar]
  • 20.Chen S, Zhao G, Miao H, Tang R, Song Y, Hu Y, Wang Z, Hou Y. MicroRNA-494 inhibits the growth and angiogenesis-regulating potential of mesenchymal stem cells. FEBS Lett. 2015;589:710–17. doi: 10.1016/j.febslet.2015.01.038. [DOI] [PubMed] [Google Scholar]
  • 21.Hromadnikova I, Kotlabova K, Ondrackova M, Kestlerova A, Novotna V, Hympanova L, Doucha J, Krofta L. Circulating C19MC microRNAs in preeclampsia, gestational hypertension, and fetal growth restriction. Mediators Inflamm. 2013;2013:186041. doi: 10.1155/2013/186041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Wang Y, Fan H, Zhao G, Liu D, Du L, Wang Z, Hu Y, Hou Y. miR-16 inhibits the proliferation and angiogenesis-regulating potential of mesenchymal stem cells in severe pre-eclampsia. FEBS J. 2012;279:4510–24. doi: 10.1111/febs.12037. [DOI] [PubMed] [Google Scholar]
  • 23.Zhao G, Zhou X, Chen S, Miao H, Fan H, Wang Z, Hu Y, Hou Y. Differential expression of microRNAs in decidua-derived mesenchymal stem cells from patients with pre-eclampsia. J Biomed Sci. 2014;21:81. doi: 10.1186/s12929-014-0081-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Welten SM, Bastiaansen AJ, de Jong RC, de Vries MR, Peters EA, Boonstra MC, Sheikh SP, La Monica N, Kandimalla ER, Quax PH, Nossent AY. Inhibition of 14q32 MicroRNAs miR-329, miR-487b, miR-494, and miR-495 increases neovascularization and blood flow recovery after ischemia. Circ Res. 2014;115:696–708. doi: 10.1161/CIRCRESAHA.114.304747. [DOI] [PubMed] [Google Scholar]
  • 25.In’t Anker PS, Scherjon SA, Kleijburg-van der Keur C, de Groot-Swings GM, Claas FH, Fibbe WE, Kanhai HH. Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta. Stem Cells. 2004;22:1338–45. doi: 10.1634/stemcells.2004-0058. [DOI] [PubMed] [Google Scholar]
  • 26.Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25:402–08. doi: 10.1006/meth.2001.1262. [DOI] [PubMed] [Google Scholar]
  • 27.Murphy MS, Casselman RC, Tayade C, Smith GN. Differential expression of plasma microRNA in preeclamptic patients at delivery and 1 year postpartum. Am J Obstet Gynecol. 2015;213:361–67. doi: 10.1016/j.ajog.2015.05.013. [DOI] [PubMed] [Google Scholar]
  • 28.Terasawa K, Ichimura A, Sato F, Shimizu K, Tsujimoto G. Sustained activation of ERK1/2 by NGF induces microRNA-221 and 222 in PC12 cells. FEBS J. 2009;276:3269–76. doi: 10.1111/j.1742-4658.2009.07041.x. [DOI] [PubMed] [Google Scholar]
  • 29.Hafner M, Landthaler M, Burger L, Khorshid M, Hausser J, Berninger P, Rothballer A, Ascano M Jr, Jungkamp AC, Munschauer M, Ulrich A, Wardle GS, Dewell S, Zavolan M, Tuschl T. Transcriptome-wide identification of RNA-binding protein and microRNA target sites by PAR-CLIP. Cell. 2010;141:129–41. doi: 10.1016/j.cell.2010.03.009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Iriyama T, Wang W, Parchim NF, Song A, Blackwell SC, Sibai BM, Kellems RE, Xia Y. Hypoxiaindependent upregulation of placental hypoxia inducible factor-1alpha gene expression contributes to the pathogenesis of preeclampsia. Hypertension. 2015;65:1307–15. doi: 10.1161/HYPERTENSIONAHA.115.05314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Bosco C, Diaz E, Gutierrez R, Gonzalez J, Parra-Cordero M, Rodrigo R, Barja PY. Placental hypoxia developed during preeclampsia induces telocytes apoptosis in chorionic villi affecting the maternal-fetus metabolic exchange. Curr Stem Cell Res Ther. 2016;11:420–5. doi: 10.2174/1574888x10666150202144855. [DOI] [PubMed] [Google Scholar]
  • 32.Perez-Sepulveda A, Monteiro LJ, Dobierzewska A, Espana-Perrot PP, Venegas-Araneda P, Guzman-Rojas AM, Rodrigo RS, Barja PY. Placental aromatase is deficient in placental ischemia and preeclampsia. PLoS One. 2015;10:e139682. doi: 10.1371/journal.pone.0139682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Jauniaux E, Burton GJ. Pathophysiology of histological changes in early pregnancy loss. Placenta. 2005;26:114–23. doi: 10.1016/j.placenta.2004.05.011. [DOI] [PubMed] [Google Scholar]
  • 34.Norwitz ER. Defective implantation and placentation: laying the blueprint for pregnancy complications. Reprod Biomed Online. 2006;13:591–99. doi: 10.1016/s1472-6483(10)60649-9. [DOI] [PubMed] [Google Scholar]
  • 35.Sharp AN, Heazell AE, Crocker IP, Mor G. Placental apoptosis in health and disease. Am J Reprod Immunol. 2010;64:159–69. doi: 10.1111/j.1600-0897.2010.00837.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Li J, Chen Z, Zhou X, Shi S, Qi H, Baker PN, Zhang H. Imbalance between proliferation and apoptosis-related impaired GPR30 expression is involved in preeclampsia. Cell Tissue Res. 2016;366:499–508. doi: 10.1007/s00441-016-2466-y. [DOI] [PubMed] [Google Scholar]
  • 37.Bosco CB, Diaz EG, Gutierrez RR, Gonzalez JM, Parra-Cordero M, Rodrigo RS, Barja PY. Placental hypoxia developed during preeclampsia induces telocytes apoptosis in chorionic villi affecting the maternal-fetus metabolic exchange. Curr Stem Cell Res Ther. 2016;11:420–25. doi: 10.2174/1574888x10666150202144855. [DOI] [PubMed] [Google Scholar]
  • 38.Zou Y, Zuo Q, Huang S, Yu X, Jiang Z, Zou S, Fan M, Sun L. Resveratrol inhibits trophoblast apoptosis through oxidative stress in preeclampsia-model rats. Molecules. 2014;19:20570–79. doi: 10.3390/molecules191220570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Fang M, Du H, Han B, Xia G, Shi X, Zhang F, Fu Q, Zhang T. Hypoxia-inducible microRNA-218 inhibits trophoblast invasion by targeting LASP1: Implications for preeclampsia development. Int J Biochem Cell Biol. 2017;87:95–103. doi: 10.1016/j.biocel.2017.04.005. [DOI] [PubMed] [Google Scholar]
  • 40.Korkes HA, De Oliveira L, Sass N, Salahuddin S, Karumanchi SA, Rajakumar A. Relationship between hypoxia and downstream pathogenic pathways in preeclampsia. Hypertens Pregnancy. 2017;36:145–50. doi: 10.1080/10641955.2016.1259627. [DOI] [PubMed] [Google Scholar]
  • 41.Qu H, Qu L, Li X, Pan X. Overexpressed HO-1 is associated with reduced STAT3 activation in preeclampsia placenta and inhibits STAT3 phosphorylation in placental JEG-3 cells under hypoxia. Arch Med Sci. 2016 doi: 10.5114/aoms.2016.63261. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from International Journal of Clinical and Experimental Pathology are provided here courtesy of e-Century Publishing Corporation

RESOURCES