Zhang Xue Mei, Xiong Xi, Tong Chao, Li Qin, Huang Shuai, Li Qing Shu, Liu Ya Ming, Li Hai Ying, Baker Phillip, Shan Nan, Qi Hong Bo
Department of Obstetrics, The First Affiliated Hospital, Chongqing Medical University, Chongqing, China.
Department of Pathology, Chongqing Medical University, Chongqing, China.
Cell Physiol Biochem. 2018;51(5):2030-2040. doi: 10.1159/000495822. Epub 2018 Dec 6.
BACKGROUND/AIMS: Preeclampsia (PE) is a gestational disorder defined as hypertension and proteinuria, which is deemed a major cause of maternal and neonatal mortality and morbidity worldwide. The aim of this study was to investigate the expression patterns of placental laminin (LN)-α5 expression in normal and PE pregnancies, as well as evaluating the effects of LN-α5 on trophoblast proliferation, apoptosis, and invasion.
LN-α5 expression levels were examined by reverse-transcriptase polymerase chain reaction (RT-PCR), and further confirmed by western blotting and immunofluorescence staining. Cell proliferation and apoptosis were measured by CCK-8 assay and flow cytometry. Cell invasion was assessed by matrigel-based transwell assay. LN-α5 DNA methylation in placentas was determined by bisulfite sequencing PCR (BSP).
LN-α5 expression levels in PE placentas were significantly lower than that of normal pregnancies. Deficiency in LN-α5 expression resulted in decreased trophoblast proliferation and invasion but increased cell apoptosis, meanwhile, PI3K/AKT/mTOR signaling pathway was impaired by LN-α5 silencing. LN-α5 promoter methylation didn't show significant difference between PE and normal placentas.
LN-α5 downregulation is associated with PE placenta and impairs trophoblast viability and invasiveness, which could be a causative factor of PE pathogenesis.
背景/目的：子痫前期（PE）是一种妊娠期疾病，定义为高血压和蛋白尿，被认为是全球孕产妇和新生儿死亡及发病的主要原因。本研究的目的是调查正常妊娠和子痫前期妊娠中胎盘层粘连蛋白（LN）-α5的表达模式，并评估LN-α5对滋养细胞增殖、凋亡和侵袭的影响。
通过逆转录聚合酶链反应（RT-PCR）检测LN-α5表达水平，并通过蛋白质印迹法和免疫荧光染色进一步证实。采用CCK-8法和流式细胞术检测细胞增殖和凋亡。通过基于基质胶的Transwell实验评估细胞侵袭。通过亚硫酸氢盐测序PCR（BSP）测定胎盘组织中LN-α5的DNA甲基化情况。
子痫前期胎盘组织中LN-α5表达水平显著低于正常妊娠。LN-α5表达缺失导致滋养细胞增殖和侵袭减少，但细胞凋亡增加，同时，LN-α5沉默会损害PI3K/AKT/mTOR信号通路。子痫前期胎盘组织与正常胎盘组织中LN-α5启动子甲基化无显著差异。
LN-α5下调与子痫前期胎盘有关，并损害滋养细胞活力和侵袭能力，这可能是子痫前期发病机制的一个致病因素。
