定量检测子痫前期孕妇血浆中超甲基化的RASSF1A基因

Quantitative detection of the hypermethylated RASSF1A gene in maternal plasma of pre-eclampsia

目的探讨子痫前期孕妇血浆中超甲基化的Ras相关区域家族1A（ras association domain family 1A，RASSF1A）基因的水平变化及应用价值。方法选择晚期妊娠子痫前期患者60例为实验组，其中轻度和重度子痫前期各30例；正常晚期孕妇60名为对照组。提取血浆游离DNA，经甲基化敏感的限制性内切酶处理，实时定量检测酶切前、后RASSF1A基因的浓度，同时检测β肌动蛋白（β-actin）基因以确保酶的完全消化。结果子痫前期孕妇血浆中超甲基化的RASSF1A基因含量为正常妊娠组的3．31倍。轻、重度子痫前期患者浓度有差异，中位数分别为1659．00copies／mL及2036．50copies／mL（P〈0．05）。结论子痫前期孕妇血浆中的超甲基化RASSF1A基因含量明显升高，且浓度水平与子痫前期的严重程度相关。

Objective To investigate the level of hypermethylated ras association domain family 1A （RASSF1A）gene in maternal plasma of pre-eclampsia and its clinical value. Methods Sixty pre-eclampsia women including 30 mild and 30 severe cases were selected, 60 women with normal pregnancy were studied as control. Free DNA from plasma samples was extracted, fluorescence quantitative polymerase chain reaction （FQ PCR） was used to detect the concentrations of RASSF1A gene before and after methylationsensitive restriction digestion. Meanwhile, β-actin gene was detected as a control to confirm complete enzyme digestion. Results The median concentration of hypermethylated RASSF1A gene was 3. 31-fold higher in samples from pre-eclamptic pregnancies than that in controls. There was significant difference between the mild and severe pre-eclamptic subjects （P〈0. 05）, with the median concentrations of 1659 copies/mL and 2036. 50 copies/mL, respectively. Conclusion Hypermethylated RASSF1A gene in preeclampsia plasma was significantly increased and the concentrations were related to the severity of preeclampsia.