关键词: 长链非编码RNA, 胃癌高表达转录本1, 子痫前期, Wnt/β-catenin信号通路, 细胞增殖, 细胞侵袭

Abstract:

Objective： To investigate the expression of long non-coding RNA gastric carcinoma high expression transcript 1 （lncRNA GHET1） in placenta tissue of the preeclampsia （PE） patients and its effects on the proliferation， cycle progression and invasion ability of trophoblast cells， and to clarify their related mechanisms. Methods Thirty pregnant and lying-in women were divided into normal group（n=15） and PE group （n=15）. The pathomorphology of placenta tissue of the subjects in two groups were observed by HE staining.The HTR-8 cells were cultured in vitro， and transfected with over-expression lncRNA GHET1 and control sequence plasmids， and the cells were divided into control group （conventional culture）， GHET1 group （trasfected with over-expressed lncRNA GHET1 plasmid） and negative control group （transfected with negative control sequence plasmid）.Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of lncRNA GHET1 mRNA in placenta tissue of the subjects in two groups and HTR-8 cells in various groups. The proliferation rates of HTR-8 cells in various groups were measured by CCK-8 method. The percentages of HTR-8 cells at different cell cycles in various groups were detected by flow cytometry. The invasion abilities of HTR-8 cells in various groups were detected by Transwell chamber method. Western blotting method was used to detect the expression levels of cellular-myelocytomatosis viral oncogene（c-Myc）， cyclinD1 and epithelial mesenchymal transformation （EMT）-associated protein E-cadherin，and Vimentin and the expression levels of Wnt/β-catenin signaling pathway-associated proteins and ratio of phosphorylated glycogen synthase kinase 3β （p-GSK3β）/glycogen synthase kinase 3β（GSK3β） and β-catenin proteins in the HTR-8 cells in various groups. Results Compared with normal group， the villi of placenta tissue of the patients in PE group were dysplasia and the amount was decreased， the vascular distribution in villi and interstitium was disorder， the vascular wall showed fibrinoid necrosis， and the calcified area and syncytial nodule on the villi were increased. Compared with normal group， the expression level of lncRNA GHET1 mRNA in placenta tissue of the patients in PE group was significantly decreased （P<0.05）. Compared with control group， the lncRNA GHET1 mRNA expression level in the HTR-8 cells in GHET1 group was significantly increased（P<0.05）， while there was no significant difference in the indicators mentioned above in negative control group （P>0.05）. Compared with control group， the proliferation rate， the percentage of cells at S phase，and the number of invasion cells of HTR-8 cells in GHET1 group were significantly increased （P<0.05）， while there were no significant differences in the indicators mentioned above in negative control group（P>0.05）. Compared with control group， the expression levels of c-Myc， cyclinD1， Vimentin and β-catenin proteins and ratio of p-GSK3β/GSK3β in the HTR-8 cells in GHET1 group were significantly increased （P<0.05）， while the expression level of E-cadherin was significantly decreased （P<0.05）； there were no significant differences in the indicators mentioned above in negative control group （P>0.05）. Conclusion Over-expression of lncRNA GHET1 in placenta tissue of the PE patients may promote trophoblast cell proliferation， cell cycle progression， and cell invasion through Wnt/β-catenin signaling pathway， and play a role in improving the progression of PE.

Key words: Long non-coding RNA, Gastric cancinoma high expressed transcript 1, Preeclampsia, Wnt/β-catenin signaling pathway, Cell proliferation, Cell invasion