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目的:通过研究人中胚层特异性转录基因(mesoderm-specific transcript,MEST)对人绒毛滋养细胞迁移和侵袭等生物学功能的影响,探讨其在子痫前期(preeclampsia,PE)中发病的机制。方法:用免疫组化法和Western blot法检测正常和PE患者胎盘中MEST及DNA甲基转移酶(DNA methyltransferase,DNMT)的表达。并检测2组胎盘中MEST甲基化水平。用Western blot法检测缺氧/复氧(hypoxia/reoxygenation,H/R)和MEST si RNA处理人滋养细胞株HTR8/Svneo后缺氧诱导因子(hypoxia-inducible factor 1α,HIF1α)及MEST表达水平;并检测si RNA转染后对各组细胞增殖、迁移和侵袭能力的影响。结果:MEST在正常胎盘中的表达(4.524±0.586)明显高于PE胎盘(2.640±0.334)且在PE胎盘中处于高甲基化状态。H/R后HIF1α表达明显升高(3.692±0.441,7.091±0.698),而MEST表达明显降低(4.856±0.169,2.205±0.318),特异性敲低MEST后HTR-8细胞侵袭和迁移能力均明显下降(1.057±0.061,0.551±0.029)(1.113±0.087,0.477±0.034),差异均有统计学意义(P<0.05);而增殖能力无明显影响(0.170±0.010,0.158±0.010)(P>0.05)。结论:推测MEST DNA甲基化异常可能影响滋养细胞的迁移和侵袭,从而参与子痫前期的发病。
OBJECTIVE: To investigate the effect of mesoderm-specific transcript (MEST) on the biological function of human chorionic villus trophoblast migration and invasion, and to explore its mechanism in preeclampsia (PE). Methods: The expression of MEST and DNA methyltransferase (DNMT) in the placenta of normal and PE patients were detected by immunohistochemistry and Western blot. The methylation level of MEST in two groups of placenta was detected. Western blot was used to detect the hypoxia-inducible factor 1α (HIF1α) and MEST expression levels after hypoxia / reoxygenation (H / R) and MEST si RNA treatment of human trophoblast cell line HTR8 / Svneo; The effect of si RNA transfection on the proliferation, migration and invasion of cells in each group was also tested. Results: The expression of MEST in normal placenta (4.524 ± 0.586) was significantly higher than that of PE placenta (2.640 ± 0.334) and was hypermethylated in PE placenta. After H / R, the expression of HIF1α was significantly increased (3.692 ± 0.441, 7.091 ± 0.698), while the expression of MEST was significantly lower (4.856 ± 0.169, 2.205 ± 0.318). The ability of invasion and migration of HTR-8 cells was significantly lower after specific knockdown of MEST (1.057 ± 0.061,0.551 ± 0.029) (1.113 ± 0.087,0.477 ± 0.034), the difference was statistically significant (P <0.05), while the proliferation ability had no significant effect (0.170 ± 0.010,0.158 ± 0.010) (P> 0.05). Conclusion: It is speculated that abnormal methylation of MEST DNA may affect the migration and invasion of trophoblast cells and thus participate in the pathogenesis of preeclampsia.