子痫前期患者胎盘组织中抑微管装配蛋白1表达对滋养层细胞的影响及其机制

doi:10.13481/j.1671-587X.20230615

摘要/Abstract

摘要：

目的探讨子痫前期（PE）患者胎盘组织中抑微管装配蛋白1（STMN1）表达对滋养层细胞侵袭和迁移功能的影响，并阐明其相关机制。方法选取17名健康妊娠孕产妇（对照组）和15例PE患者（PE组）的胎盘组织。将人滋养层HTR-8细胞分为siRNA-STMN1组（转染siRNA-STMN1）、siRNA-NC组（转染阴性对照序列siRNA-NC）、pcDNA3.1-STMN1组（转染pcDNA3.1-STMN1）和pcDNA3.1-NC组（转染阴性对照序列pcDNA3.1-NC）。采用免疫组织化学（IHC）染色观察2组研究对象胎盘组织中STMN1和上皮-间质转化（EMT）相关蛋白表达强度，CCK-8法检测各组HTR-8细胞增殖率，细胞划痕实验检测各组HTR-8细胞迁移能力，Transwell小室实验检测各组HTR-8细胞侵袭能力，实时荧光定量PCR（RT-qPCR）法检测2组研究对象和各组HTR-8细胞中STMN1 mRNA表达水平，Western blotting检测2组研究对象和各组HTR-8细胞中STMN1、E-钙黏蛋白（E-cadherin）和N-钙黏蛋白（N-cadherin）蛋白表达水平。结果与对照组比较，PE组患者收缩压、舒张压和尿蛋白水平均明显升高（P<0.01），新生儿体质量增加（P<0.05）。IHC染色观察，与对照组比较，PE组患者胎盘绒毛组织中STMN1和E-cadherin蛋白表达强度增加，N-cadherin蛋白表达强度降低。CCK-8法检测，与siRNA-NC组比较，siRNA-STMN1组HTR-8细胞增殖率降低（P<0.05）；与pcDNA3.1-NC组比较，pcDNA3.1-STMN1组HTR-8细胞增殖率升高（P<0.05）。细胞划痕实验检测，与siRNA-NC组比较，siRNA-STMN1组HTR-8细胞迁移率降低（P<0.05）；与pcDNA3.1-NC组比较，pcDNA3.1-STMN1组HTR-8细胞迁移率升高（P<0.05）。Transwell小室实验检测，与siRNA-NC组比较，siRNA-STMN1组侵袭细胞数明显减少（P<0.01）；与pcDNA3.1-NC组比较，pcDNA3.1-STMN1组侵袭细胞数明显增加（P<0.01）。RT-qPCR法检测，与对照组比较，PE组患者胎盘组织中STMN1 mRNA表达水平降低（P<0.05）；与siRNA-NC组比较，siRNA-STMN1组HTR-8细胞中STMN1 mRNA表达水平明显降低（P<0.01）；与pcDNA3.1-NC组比较，pcDNA3.1-STMN1组HTR-8细胞中STMN1 mRNA表达水平明显升高（P<0.01）。Western blotting法检测，与对照组比较，PE组患者胎盘组织中STMN1蛋白表达水平降低（P<0.05）；与siRNA-NC组比较，siRNA-STMN1组HTR-8细胞中STMN1和N-cadherin蛋白表达水平降低（P<0.05），E-cadherin蛋白表达水平升高（P<0.05）。与pcDNA3.1-NC组比较，pcDNA3.1-STMN1组HTR-8细胞中STMN1和N-cadherin蛋白表达水平升高（P<0.05），E-cadherin蛋白表达水平降低（P<0.05）。结论 PE患者胎盘组织中STMN1表达明显减少，STMN1可能通过调控滋养层细胞的EMT进程抑制细胞增殖、细胞侵袭和细胞迁移能力，从而参与PE的发生发展。

关键词: 子痫前期, 抑微管装配蛋白1, 滋养层细胞, 细胞增殖, 细胞侵袭, 上皮-间质转化

Abstract:

Objective To discuss the effect of the expression of stathmin 1 （STMN1） in placenta tissue of the pre-eclampsia （PE） patients on the invasion and migration functions of the trophoblast cells， and to clarify the related mechanism. Methods The placenta tissues of 17 healthy pregnant women （control group） and 15 PE patients （PE group） were selected. The human trophoblast HTR-8 cells were divided into siRNA-STMN1 group （transfected with siRNA-STMN1）， siRNA-NC group （transfected with negative control sequence siRNA-NC），pcDNA3.1-STMN1 group （transfected with pcDNA3.1-STMN1）， and pcDNA3.1-NC group （transfected with negative control sequence pcDNA3.1-NC）. The expression intensities of STMN1 and epithelial-mesenchymal transition（EMT）-related proteins in placenta tissue of the patients in two groups were observed by immunohistochemistry （IHC） staining； the proliferation rates of the HTR-8 cells in various groups were detected by CCK-8 method； the migration abilities of the HTR-8 cells in various groups were detected by cell scratch experiment； the invasion abilities of the HTR-8 cells in various groups were detected by Transwell chamber experiment； the expression levels of STMN1 mRNA in placenta tissue of the patients in two groups and HTR-8 cells were detected by real-time fluorescence quantitative PCR （RT-qPCR） method； the expression levels of STMN1， E-cadherin， and N-cadherin proteins in placenta tissue of the patients in two groups and HTR-8 cells were detected by Western blotting method. Results Compared with control group， the systolic blood pressure， diastolic blood pressure， and the level of urine protein of the patients in PE group were significantly increased （P<0.01）， and the body weight of the newborn was increased （P<0.05）.The IHC staining results showed that compared with control group， the expression intensities of STMN1 and E-cadherin proteins in placenta tissue of the patients in PE group were increased， and the expression intensity of N-cadherin protein was decreased.The CCK-8 results showed that compared with siRNA-NC group， the proliferation rate of the HTR-8 cells in siRNA-STMN1 group was decreased（P<0.05）； compared with pcDNA3.1-NC group，the proliferation rate of the HTR-8 cells in pcDNA3.1-STMN1 group was increased（P<0.05）.The cell scratch experiment results showed that compared with siRNA-NC group，the migration rate of the HTR-8 cells in siRNA-STMN1 group was decreased（P<0.05）；compared with the pcDNA3.1-NC group， the migration rate of the HTR-8 cells in pcDNA3.1-STMN1 group was increased （P<0.05）. The Transwell chamber experiment results showed that compared with siRNA-NC group， the number of invasion cells in siRNA-STMN1 group was significantly decreased （P<0.01）； compared with pcDNA3.1-NC group， the number of invasion cells in pcDNA3.1-STMN1 group was significantly increased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression level of STMN1 mRNA in placenta tissue of the patients in PE group was decreased （P<0.05）； compared with siRNA-NC group， the expression level of STMN1 mRNA in the HTR-8 cells in siRNA-STMN1 group was significantly decreased （P<0.01）； compared with pcDNA3.1-NC group， the expression level of STMN1 mRNA in the HTR-8 cells in pcDNA3.1-STMN1 group was significantly increased （P<0.01）. The Western blotting results showed that compared with control group， the expression level of STMN1 protein in placenta tissue of the patients in PE group was decreased （P<0.05）. Compared with siRNA-NC group， the expression levels of STMN1 and N-cadherin proteins in the HTR-8 cells in siRNA-STMN1 group were decreased （P<0.05）， and the expression level of E-cadherin protein was increased （P<0.05）. Compared with pcDNA3.1-NC group， the expression levels of STMN1 and N-cadherin proteins in the HTR-8 cells in pcDNA3.1-STMN1 group were increased （P<0.05）， and the expression level of E-cadherin protein was decreased （P<0.05）. Conclusion The expression of STMN1 in placenta tissue of the PE patients is significantly reduced，suggesting that STMN1 may be involved in the development of PE by regulating the EMT process of the trophoblast cells and inhibiting the cell proliferation， invasion， and migration.

Key words: Preeclampsia, Stathmin 1, Trophoblast cell, Cell proliferation, Cell invasion, Epithelial-mesenchymal transition

中图分类号:

R714.25

欧曼颖,胡春霞,李跃萍. 子痫前期患者胎盘组织中抑微管装配蛋白1表达对滋养层细胞的影响及其机制[J]. 吉林大学学报(医学版), 2023, 49(6): 1519-1527.

Manying OU,Chunxia HU,Yueping LI. Effect of expression of microtubule inhibitory assembly protein 1 in placenta tissue of pre-eclampsia patients on trophoblast cells and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2023, 49(6): 1519-1527.

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