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邹艳芬, 孙丽洲. 长链非编码RNA HOTAIR调节PE滋养细胞功能的研究[J]. 四川大学学报(医学版), 2015, 46(1): 113-117.
引用本文: 邹艳芬, 孙丽洲. 长链非编码RNA HOTAIR调节PE滋养细胞功能的研究[J]. 四川大学学报(医学版), 2015, 46(1): 113-117.
ZOU Yan-fen, SUN Li-zhou. Long Noncoding RNA HOTAIR Modulates the Function of Trophoblast Cells in Pre-eclampsia[J]. Journal of Sichuan University (Medical Sciences), 2015, 46(1): 113-117.
Citation: ZOU Yan-fen, SUN Li-zhou. Long Noncoding RNA HOTAIR Modulates the Function of Trophoblast Cells in Pre-eclampsia[J]. Journal of Sichuan University (Medical Sciences), 2015, 46(1): 113-117.

长链非编码RNA HOTAIR调节PE滋养细胞功能的研究

Long Noncoding RNA HOTAIR Modulates the Function of Trophoblast Cells in Pre-eclampsia

  • 摘要: 目的 观察长链非编码RNA(lncRNA)HOX转义反录RNA(HOTAIR)在子痫前期(PE)胎盘组织中的表达及其对滋养细胞HTR-8/SVneo增殖、侵袭及凋亡能力的影响。方法 采用实时定量聚合酶链反应(qRT-PCR)法检测23例重度PE和23例正常产妇胎盘组织中HOTAIR mRNA表达水平。分别构建其封闭(si-HOTAIR)及过表达载体(pcDNA-HOTAIR),与其对照(si-NC及空载体)分别转染HTR-8/SVneo细胞24~48 h后, qRT-PCR检测细胞中HOTAIR mRNA表达水平验证干扰效率和过表达倍数。采用克隆形成实验和MTT法检测转染后细胞生长增殖能力,Transwell法检测转染后细胞迁移、侵袭能力改变。应用流式细胞技术检测转染后细胞凋亡改变, Western blot检测凋亡相关蛋白表达情况。结果 PE胎盘组织中HOTAIR水平高于正常组织(P<0.05); pcDNA-HOTAIR组细胞HOTAIRmRNA过表达倍数为51.27; si-HOTAIRHOTAIRmRNA表达被抑制超过95%,证明过表达和封闭均有效;pcDNA-HOTAIR组细胞增殖能力降低, si-HOTAIR组细胞增殖增加(P<0.05);pcDNA-HOTAIR组细胞的穿膜细胞数低于对照组,si-HOTAIR组细胞穿膜数高于对照组(P<0.05);pcDNA-HOTAIR组细胞凋亡增加,si-HOTAIR组凋亡被抑制(P<0.05);pcDNA-HOTAIR组促凋亡蛋白Caspase-3表达增加,而si-HOTAIR组降低;抑凋亡蛋白 BCL-2与之相反(P<0.05)。结论 HOTAIR可能通过抑制滋养细胞的增殖和侵袭能力,加速滋养细胞凋亡,从而参与PE的发生发展。

     

    Abstract: Objective To investigate the expression of long noncoding RNA (lncRNA) HOTAIR in pre-eclampsia (PE) placentas and its effect on trophoblast cells proliferation, invasion, and apoptosis. Methods The expression of HOTAIR was determined by qPCR for 23 severe PE and 23 normal pregnant women. qRT-PCR was used to detect the efficiency of overexpression and knock-down after the HTR-8/SVneo cells were transfected with HOTAIR overexpression and siRNAs targeting HOTAIR for 24-48 h, respectively. MTT and colony formation assays were used to test the proliferation of HTR-8/SVneo cells transfected. Transwell assay was used to show the migration and invasion ability of HTR-8/SVneo cells transfected. Flow cytometry assay was used to detect the cell apoptosis rate after treatment. Western blot assay was applied to detect the expression level of apoptotic proteins Caspase-3 and BCL-2. Results The level of HOTAIR in severe pre-eclampsia groups was significantly increased compared to normal pregnant placentas (P <0.05). The expression of HOTAIR in HTR-8/SVneo cells after transfected with pcDNA-HOTAIR was 51.27-fold than that of control; while inhibition of HOTAIR was more than 95% in si-HOTAIR than that of control. According to the MTT and colony formation assays, we found that cells proliferation rate of cells were significantly decreased in overexpression HOTAIR group while increased in si-HOTAIR group when compared with control, respectively. The transwell assay showed that the invasive capacity of HTR-8/SVneo cells in cells transfected with pcDNA-HOTAIR decreased, while increased in si-HOTAIR transfected cells when compared with that of control. The apoptosis rate in cells transfected with HOTAIR overexpression was apparently more than that of control, while less in cells treated with si-HOTAIR. Western blot assay showed that the Caspase-3 showed an obvious increase in HOTAIR overexpression group while decreased in si-HOTAIR group. And BCL-2 presented an opposite trend. Conclusion HOTAIR is probably involved in the onset of preeclampsia by regulating proliferation, invasion and apoptosis of trophoblast cells.

     

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