[1]王 维a,唐 静b,杨永红,等.妊娠期高血压疾病患者胎盘组织叶酸代谢基因MTR 的表达水平及其作用机制研究[J].现代检验医学杂志,2022,37(04):134-137+173.[doi:10.3969/j.issn.1671-7414.2022.04.026]
　WANG Weia,TANG Jingb,YANG Yong-hong,et al.Expression Level and Effects of Folate Metabolism-related Gene MTR in Placenta of Patients with Hypertensive Disorder Complicating Pregnancy[J].Journal of Modern Laboratory Medicine,2022,37(04):134-137+173.[doi:10.3969/j.issn.1671-7414.2022.04.026]
点击复制

妊娠期高血压疾病患者胎盘组织叶酸代谢基因MTR 的表达水平及其作用机制研究()
分享到：

《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年04期
页码:
134-137+173
栏目:
论著
出版日期:
2022-07-15

文章信息/Info

Title:
Expression Level and Effects of Folate Metabolism-related Gene MTR in Placenta of Patients with Hypertensive Disorder Complicating Pregnancy
文章编号:
1671-7414（2022）04-134-06
作者:
王 维1a; 唐 静1b; 杨永红2; 薛 峰2
1. 成都市龙泉驿区妇幼保健院 a. 检验科；b. 妇产科，成都 610100;2. 四川大学华西医院龙泉医院检验科，成都 610100
Author(s):
WANG Wei1a;  TANG Jing1b;  YANG Yong-hong2;  XUE Feng2
1a.Department of Laboratory Medicine；1b.Department of Obstetrics and Gynecology, Longquanyi District Maternal and Child Health Hospital, Chengdu 610100, China; 2.Department of Laboratory Medicine, West China Longquan Hospital, Sichuan University, Chengdu 610100, China
关键词:
妊娠期高血压; 胎盘组织; 叶酸代谢; 甲硫氨酸合成酶
分类号:
R714.246；Q786
DOI:
10.3969/j.issn.1671-7414.2022.04.026
文献标志码:
A
摘要:
目的 研究叶酸代谢相关基因甲硫氨酸合成酶(methionine synthase，MTR) 在妊娠期高血压疾病患者胎盘组织中的差异表达以及对于人绒毛滋养层细胞HTR8 迁移及侵袭能力调控的机制研究。方法 2018 年1 月～ 2020 年12 月在成都市龙泉驿区妇幼保健院分娩的30 例妊娠期高血压，28 例子痫前期和20 例重度子痫前期患者及25 例正常产妇作为研究对象，分娩时采集研究对象胎盘组织。使用实时荧光定量PCR(real-time quantitative PCR detecting system,qPCR)及蛋白质印迹法(western blotting,WB) 检测了各组胎盘组织中MTR，上皮型钙黏附蛋白(E-Cadherin,E-Cad)，锌指E 盒结合蛋白-1(zinc finger E-box binding protein1,ZEB1) 基因转录及蛋白表达情况。进一步通过siRNA 敲减人绒毛滋养层细胞HTR8 中MTR 基因的表达，研究MTR 对于E-Cadherin 和ZEB1 基因的调控作用。通过Transwell 迁移及侵袭实验验证敲减MTR 后对于HTR8 细胞迁移和侵袭的影响。结果 与对照组相比，随着妊娠期高血压疾病分期的进展，子痫前期与重度子痫前期患者胎盘组织中E-Cadherin 的表达增加（1.14±0.35, 1.53±0.41 vs 1.00±0.30），胎盘组织中MTRmRNA 的表达量逐渐增加（1.72±0.17, 2.58±0.13 vs 1.00±0.33），妊娠期高血压疾病患者胎盘组织中ZEB1 的表达量下降（0.48±0.10, 0.13±0.06 vs 1.00±0.22），差异具有统计学意义（F=13.28，67.1，65.41，均 P ＜ 0.01）。随着疾病分期进展，MTR 及E-Cadherin 的蛋白表达量明显增加，而ZEB1 的蛋白表达量明显降低。siRNA 可以显著抑制HTR8中MTR 基因mRNA 的表达，敲低MTR 基因后可显著抑制上皮相关标志物E-Cadherin 基因mRNA 的表达, 可以显著促进间充质相关标志物ZEB1 基因mRNA 的表达，差异具有统计学意义（t=5.906，6.715，9.777, 均 P ＜ 0.01）。敲减HTR8 细胞中MTR 基因可显著促进迁移能力，侵袭实验结果显示敲减MTR 基因可促进HTR8 细胞的侵袭能力，差异具有统计学意义（t=6.241，22.37, 均 P ＜ 0.01）。结论 妊娠期高血压疾病患者胎盘组织中MTR 基因表达上调， MTR可通过调节HTR8 细胞EMT 相关基因E-Cadherin，ZEB1 表达，从而抑制其迁移及侵袭能力。MTR 可能成为妊娠期高血压疾病潜在的治疗靶点之一。
Abstract:
Objective To study the differential expression of folate metabolism-related gene methionine synthase (MTR) in the placenta tissue of patients with pregnancy-induced hypertension and its regulation of the migration and invasion of human chorionic trophoblast cells HTR8. Methods From January 2018 to December 2020, 30 cases of pregnancy-induced hypertension, 28 cases of preeclampsia, 20 cases of severe preeclampsia and 25 normal parturients who gave birth at the Longquanyi District Maternity and Child Health Hospital of Chengdu were the subjects of study. The placental tissue of the research subject was collected at the time. The transcription and protein expression of MTR, E-Cadherin and ZEB1 genes in each group of placental tissues were detected by qPCR and western blotting. Furthermore, siRNA was used to knock down the expression of MTR gene in human chorionic trophoblast cells HTR8 to study the regulation effect of MTR on E-Cadherin and ZEB1 genes. Transwell migration and invasion experiments were conducted to verify the effect of knocking down MTR on the migration and invasion of HTR8 cells. Results Compared with the control group, with the progress of the staging of hypertension in pregnancy, the expression of E-Cadherin （1.14±0.35, 1.53±0.41 vs 1.00±0.30）in the placenta of patients with preeclampsia and severe preeclampsia increased, and the expression of MTR mRNA in the placenta tissue gradually increased（1.72±0.17, 2.58±0.13 vs 1.00±0.33）. Pregnancy the expression of ZEB1 in the placenta of patients with hypertensive disease decreased（0.48±0.10, 0.13±0.06 vs 1.00±0.22）, and the differences were statistically significant （F=13.28，67.1，65.41，all P ＜ 0.01）. As the stage of the disease progressed, the protein expression of MTR and E-Cadherin increased significantly, while the protein expression of ZEB1 decreased significantly. siRNA could significantly inhibit the expression of MTR gene mRNA in HTR8. Knockdown of MTR gene could significantly inhibit the expression of epithelialrelated marker E-Cadherin gene mRNA, and could significantly promote the expression of mesenchymal-related marker ZEB1 gene mRNA, the differences were statistically significant（t=5.906, 6.715, 9.777, all P ＜ 0.01）. Knockdown of MTR gene in HTR8 cells can significantly promote the migration ability，the results of invasion experiments show that knockdown of MTR gene can promote the invasion ability of HTR8 cells, the differences were statistically significant （t=6.241, 22.37, all P ＜0.01）. Conclusion The expression of MTR gene in the placenta tissue of patients with hypertension in pregnancy was up-regulated, while the expression of EMT-related genes changes. MTR could inhibit the migration and invasion ability of HTR8 cells by regulating the expression of E-Cadherin and ZEB1 related genes of HTR8 cells. MTR may become one of the potential therapeutic targets of hypertension in pregnancy.

参考文献/References:

[1] CHAPPELL L C, CLUVER C A, KINGDOM J, et al.Pre-eclampsia[J]. Lancet, 2021, 398(10297):341-354.
[2] IVES C W, SINKEY R, RAJAPREYAR I, et al. Preeclampsia-pathophysiology and clinical presentations: JACC state-of-the-Art review[J]. Journal of the American College of Cardiology, 2020, 76(14):1690-1702.
[3] PHIPPS E A, THADHANI R, BENZING T, et al.Pre-eclampsia: pathogenesis, novel diagnostics and therapies[J]. Nature Reviews Nephrology, 2019, 15(5):275-289.
[4] LYALL F, ROBSON S C, BULMER J N. Spiral artery remodeling and trophoblast invasion in preeclampsia and fetal growth restriction: relationship to clinical outcome[J]. Hypertension, 2013, 62(6): 1046-1054.
[5] OSUNKALU V O, TAIWO I A, MAKWE C C,et al.Methylene tetrahydrofolate reductase and methionine synthase gene polymorphisms as genetic determinants of pre-eclampsia[J]. Pregnancy Hypertens, 2020, 20:7-13.
[6] DUCKER G S, RABINOWITZ J D. One-carbon metabolism in health and disease[J]. Cell Metabolism,2017, 25(1): 27-42.
[7] TRIPODI F, BADONE B, VESCOVI M, et al.Methionine supplementation affects metabolism and reduces tumor aggressiveness in liver cancer cells[J].Cells (Basel, Switzerland), 2020, 9(11): 2491.
[8] COPPOLA A, ILISSO C P, STELLAVATO A, et al. S-Adenosylmethionine inhibits cell growth and migration of triple negative breast cancer cells through upregulating miRNA-34c and miRNA-449a[J].International Journal of Molecular Sciences, 2020,22(1): 286.
[9] MOSCA L, MINOPOLI M, PAGANO M, et al. Effects of S-adenosyl-L-methionine on the invasion and migration of head and neck squamous cancer cells and analysis of the underlying mechanisms[J]. International Journal of Oncology, 2020, 56(5): 1212-1224.
[10] SEREMAK-MROZIKIEWICZ A, BOGACZ A, BARTKOWIAK-WIECZOREK J, et al. The importance of MTHFR, MTR, MTRR and CSE expression levels in Caucasian women with preeclampsia[J]. Eur J Obstet Gynecol Reprod Biol,2015, 188:113-117.
[11] SHIN J A, KIM Y J, PARK H, et al. Localization of folate metabolic enzymes, methionine synthase and 5,10-methylenetetrahydrofolate reductase in human placenta[J]. Gynecologic and Obstetric Investigation,2014, 78(4): 259-265.
[12] KIM Y R, KANG S H, HONG S H. Effects of methionine synthase and methionine synthase reductase polymorphisms on hypertension susceptibility[J].Genes Genomics, 2020 , 42(4):477-483.
[13] ALSO-RALLO E, LOPEZ-QUESADA E, URREIZTI R, et al. Polymorphisms of genes involved in homocysteine metabolism in preeclampsia and in uncomplicated pregnancies[J]. European Journal of Obstetrics & Gynecology and Reproductive Biology,2005, 120(1): 45-52.
[14] SEREMAK-MROZIKIEWICZ A, BOGACZ A,DEKA-PAWLIK D, et al. The polymorphisms of methionine synthase (MTR) and methionine synthase reductase (MTRR) genes in pathogenesis of preeclampsia[J]. The Journal of Maternal-fetal & Neonatal Medicine, 2017, 30(20): 2498-2504.
[15] E DAVIES J, POLLHEIMER J, YONG H E, et al.Epithelial-mesenchymal transition during extravillous trophoblast differentiation[J]. Cell Adh Migr, 2016,10(3):310-321.
[16] SU M T, TSAI P Y, WANG C Y, et al. Aspirin facilitates trophoblast invasion and epithelial-mesenchymal transition by regulating the miR-200-ZEB1 axis in preeclampsia[J]. Biomedicine & Pharmacotherapy,2021, 139: 111591.
[17] WANG Rongl i , ZOU Li , YANG Xinyuan. MicroRNA-210/ long non-coding RNA MEG3 axis inhibits trophoblast cell migration and invasion by suppressing EMT process[J]. Placenta, 2021, 109: 64-71.
[18] 李黎, 王绒, 郭进京. 孕早期补充叶酸与先兆子痫发生危险性的关系[J]. 现代检验医学杂志, 2014,29(3):127-128, 130. LI Li, WANG Rong, GUO Jinjing. Risk of preeclampsia in relation folic acid supplementation during the first trimester pregnancy [J]. Journal of Modern Laboratory Medicine,2014,29(3):127-128, 130.
[19] 张燕, 王伟红, 郝万鹏. 乌鲁木齐汉族妇女亚甲基四氢叶酸还原酶基因变异与血清叶酸和同型半胱氨酸水平的相关性研究[J]. 现代检验医学杂志, 2019,34(5)：98-100, 105. ZHANG Yan, WANG Weihong, HAO Wanpeng. Study on the correlation between the variation of Methylenetetrahydrofolate reductase gene and Folate and Homocysteine in Urumqi Han women [J]. Journal of Modern Laboratory Medicine,2019,34(5):98-100, 105.
[20] HUANG Xiaowen, ZHAO Qinglin, LI Dankang, et al. Association between gene promoter methylation of the one-carbon metabolism pathway and serum folate among patients with hyperhomocysteinemia[J].European Journal of Clinical Nutrition, 2020, 74(12):1677-1684.

相似文献/References:

[1]王 碧,陈 霏.尿液KIM-1 及血浆PAPP-A 水平检测与妊娠期高血压疾病患者早期肾损伤的相关性研究[J].现代检验医学杂志,2020,35(04):79.[doi:10.3969/j.issn.1671-7414.2020.04.019]
　WANG Bi,CHEN Fei.Correlation between the Levels of Urinary KIM-1 and Plasma PAPP-A and Early Renal Injury in Patients with Pregnancy Induced Hypertension[J].Journal of Modern Laboratory Medicine,2020,35(04):79.[doi:10.3969/j.issn.1671-7414.2020.04.019]

备注/Memo

备注/Memo:
作者简介： 王维（1987-），女，本科，主管技师，研究方向：妊娠并发症以及出生缺陷相关疾病预防与控制，E-mail：shenxie70868@163.com。
通讯作者：唐静（1973-），女，本科，副主任医师，研究方向：围产期保健，E-mail：bbb124533@163.com。
更新日期/Last Update: 1900-01-01