miR-200及其甲基化调节在人早产子宫平滑肌收缩机制中的作用研究
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摘要
目的探讨miR-200及其甲基化调节在早产子宫平滑肌收缩机制中的作用。方法收集2014年1月-2015年12月该院收治的6例早产临产剖宫产孕妇子宫平滑肌作为病例组,6例未临产早产孕妇子宫平滑肌作为对照组。BSP方法检测两组miR-200启动子区域甲基化水平; Real-PCR比较两组miR-200家族表达差异,应用Dot-blot方法检测两组整体子宫平滑肌甲基化差异。结果 miR-200家族在早产临产与未临产子宫平滑肌表达均呈高甲基化状态,但临产未临产miR-200家族表达无明显差异。早产临产子宫平滑肌较早产未临产子宫平滑肌整体基因组呈低甲基化状态;结论早产子宫平滑肌基因组整体呈低甲基化状态,miR-200家族可能不参与早产子宫平滑肌收缩机制的调节。
Objective To explore the role of miR-200 and its methylation in contractile mechanism of human premature uterine smooth muscle. Methods From January 2014 to December 2015,the specimens of uterine smooth muscle of 6 cases of premature labor treated in Obstetrics and Gynecology Hospital of Fudan Unviersity were collected as case group,and the specimens of uterine smooth muscle of 6 cases of normal labor were selected as control group. BSP method was used to detect the methylation levels of miR-200 promoter region. Realtime PCR was used to compare the difference of miR-200 family expression. Dot-blot method was used to detect the difference of overall uterine smooth muscle methylation between the two groups. Results The methylation level of miR-200 family in uterine smooth muscle of cases of premature labor and normal labor were high,but there was no statistically significant difference in the expression of miR-200 family.Compared with cases of normal labor,methylation level of miR-200 family in uterine smooth muscle of cases of premature labor was lower.Conclusion The whole genome of premature uterine smooth muscle shows a low methylation state,miR-200 family may not be involved in the regulation of uterine smooth muscle contraction mechanism in premature labor.
Objective To explore the role of miR-200 and its methylation in contractile mechanism of human premature uterine smooth muscle. Methods From January 2014 to December 2015,the specimens of uterine smooth muscle of 6 cases of premature labor treated in Obstetrics and Gynecology Hospital of Fudan Unviersity were collected as case group,and the specimens of uterine smooth muscle of 6 cases of normal labor were selected as control group. BSP method was used to detect the methylation levels of miR-200 promoter region. Realtime PCR was used to compare the difference of miR-200 family expression. Dot-blot method was used to detect the difference of overall uterine smooth muscle methylation between the two groups. Results The methylation level of miR-200 family in uterine smooth muscle of cases of premature labor and normal labor were high,but there was no statistically significant difference in the expression of miR-200 family.Compared with cases of normal labor,methylation level of miR-200 family in uterine smooth muscle of cases of premature labor was lower.Conclusion The whole genome of premature uterine smooth muscle shows a low methylation state,miR-200 family may not be involved in the regulation of uterine smooth muscle contraction mechanism in premature labor.
引文
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[16]Burris HH,Collins JW Jr.Race and preterm birth--the case for epigenetic inquiry[J].Ethn Dis,2010,20(3):296-299.
[17]Zhang Q,Wang Y,Xin X,et al.Effect of folic acid supplementation on preterm delivery and small for gestational age births:a systematic review and meta-analysis[J].Reprod Toxicol,2017,67:35-41.
[18]Chen X,Bai G,Scholl TO.Spontaneous preterm delivery,particularly with reduced fetal growth,is associated with DNA hypomethylation of tumor related genes[J].J Pregnancy Child Health,2016,3(1):40.
[2]Zakar T,Mesiano S.How does progesterone relax the uterus in pregnancy?[J].N Engl J Med,2011,364(10):972-973.
[3]Renthal NE.miR-200 family and targets,ZEB1 and ZEB2,modulate uterine quiescence and contractility during pregnancy and labor[J].Proc Natl Acad Sci U S A,2010,107(48):20828-20833.
[4]Zhang N,Liu Y,Wang Y,et al.Decitabine reverses TGF-beta1-induced epithelial-mesenchymal transition in non-small-cell lung cancer by regulating miR-200/ZEB axis[J].Drug Des Devel Ther,2017,11:969-983.
[5]Vrba L,Jensen TL,Garbe JC,et al.Role for DNA methylation in the regulation of miR-200c and miR-141 expression in normal and cancer cells[J].PLoS One,2010,5(1):e8697.
[6]Li A.Pancreatic cancers epigenetically silence SIP1 and hypomethylate and overexpress miR-200a/200b in association with elevated circulating miR-200a and miR-200b levels[J].Cancer Res,2010,70(13):5226-5237.
[7]Neves R,Scheel C,Weinjhold S,et al.Role of DNA methylation in miR-200c/141 cluster silencing in invasive breast cancer cells[J].BMC Res Notes,2010,3:219.
[8]Wiklund ED,Bramsen JB,Hulf T,et al.Coordinated epigenetic repression of the miR-200 family and miR-205 in invasive bladder cancer[J].Int J Cancer,2011,128(6):1327-1334.
[9]Hassan SS.The molecular basis for sonographic cervical shortening at term:identification of differentially expressed genes and the epithelial-mesenchymal transition as a function of cervical length[J].Am J Obstet Gynecol,2010,203(5):472.e1-472.e14.
[10]Faupel-Badger JM,Fichorova RN,Allred EN,et al.Cluster analysis of placental inflammatory proteins can distinguish preeclampsia from preterm labor and premature membrane rupture in singleton deliveries less than 28 weeks of gestation[J].Am J Reprod Immunol,2011,66(6):488-494.
[11]Cook JR,Maclntyre DA,Samara E,et al.Exogenous oxytocin modulates human myometrial microRNAs[J].Am J Obstet Gynecol,2015,213(1):65.e1-65.e9.
[12]Jimenez PT,Mainigi MA,World RA,et al.miR-200 Regulates Endometrial Development During Early Pregnancy[J].Mol Endocrinol,2016,30(9):977-987.
[13]Li Y.Epigenetically deregulated miR-200c is involved in a negative feedback loop with DNMT3a in gastric cancer cells[J].Oncol Rep,2016,36(4):2108-2116.
[14]Mitsuya K.Epigenetics of human myometrium:DNA methylation of genes encoding contraction-associated proteins in term and preterm labor[J].Biol Reprod,2014,90(5):98.
[15]Bukowski R,Malone FD,Porter FT,et al.Preconceptional folate supplementation and the risk of spontaneous preterm birth:a cohort study[J].PLoS Med,2009,6(5):e1000061.
[16]Burris HH,Collins JW Jr.Race and preterm birth--the case for epigenetic inquiry[J].Ethn Dis,2010,20(3):296-299.
[17]Zhang Q,Wang Y,Xin X,et al.Effect of folic acid supplementation on preterm delivery and small for gestational age births:a systematic review and meta-analysis[J].Reprod Toxicol,2017,67:35-41.
[18]Chen X,Bai G,Scholl TO.Spontaneous preterm delivery,particularly with reduced fetal growth,is associated with DNA hypomethylation of tumor related genes[J].J Pregnancy Child Health,2016,3(1):40.