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长链非编码RNA母系印记基因3对人类绒毛发育的作用
丁海遐,马妍,盛菲,纪逸萱,李文*
0
(第二军医大学长征医院生殖医学中心, 上海 200003
*通信作者)
摘要:
目的 研究长链非编码RNA(lncRNA)母系印记基因3(MEG3)对人类流产绒毛发育的影响,并探讨其分子机制。方法 收集自然流产及人工流产各15例患者的绒毛组织,使用免疫组化方法检测其凋亡因子Bax及凋亡抑制因子Bcl-2的表达,使用qPCR法检测MEG3的表达。在人滋养细胞系HTR-8/SVneo细胞中过表达MEG3后,利用qPCR法检测MEG3的表达,利用细胞侵袭实验比较过表达组与对照组细胞的侵袭能力。结果 人工流产绒毛组织中Bax的表达低于自然流产绒毛,而Bcl-2的表达高于自然流产绒毛(P<0.01)。人工流产绒毛组织中的MEG3的表达低于自然流产(P<0.01)。人滋养层细胞系HTR-8/SVneo细胞中过表达MEG3后,HTR-8/SVneo细胞侵袭能力降低(P<0.01)。结论 lncRNA MEG3可调控滋养层细胞的凋亡及侵袭能力,影响人类绒毛的发育。
关键词:  长链非编码RNA  绒毛膜绒毛  自然流产  人工流产  母系印记基因3  细胞凋亡  侵袭
DOI:10.16781/j.0258-879x.2017.02.0183
投稿时间:2016-07-26修订日期:2016-12-19
基金项目:军队创新重点项目(13CXZ006).
Role of long non-coding RNAs maternally imprinted genes 3 in human villi development
DING Hai-xia,MA Yan,SHENG Fei,JI Yi-xuan,LI Wen*
(Center of Reproductive Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
*Corresponding author)
Abstract:
Objective To study the effect of long non-coding RNAs (lncRNAs) maternally imprinted genes 3 (MEG3) on human abortion villi development and to explore the related molecular mechanisms. Methods We collected the villi samples from 15 spontaneous abortion (SA) and 15 induced abortion (IA) patients. Immunohistochemistry was applied to detect the expressions of apoptosis factor Bax and apoptosis inhibitory factor Bcl-2 in villi samples. Real-time quantitative polymerase chain reaction (qPCR) was used to analyze the levels of MEG3 of villi samples. Overexpression of MEG3 in human trophoblast cell line HTR-8/SVneo was identified by qPCR; the invasion ability of HTR-8/SVneo cells was examined by matrigel invasion assay in MEG3 overexpression and control groups. Results Immunohistochemistry showed that the expression of Bax in IA group was lower than that in SA group, while the expression of Bcl-2 was higher (P<0.01). The level of MEG3 in IA group was significantly higher than that in SA group (P<0.01). The expression of MEG3 was obviously increased and invasion ability was inhibited in MEG3 overexpressed HTR-8/SVneo cells (P<0.01). Conclusion LncRNAs MEG3 may regulate the apoptosis and invasive ability of bizarre trophoblastic cells and influence on the development of human villi.
Key words:  long noncoding RNAs  chorionic villi  spontaneous abortion  induced abortion  maternally imprinted genes 3  apoptosis  invasion