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目的研究胰岛素样生长因子2(insulin like growth factor2,Igf2)基因的印迹状态及其表达在早期自然流产中的作用。方法选择2010年2月至2011年2月在中国医科大学附属盛京医院节育门诊就诊孕妇60例,采用PCR结合限制性片段长度多态性分析(restriction fragment length polymorphism,RFLP)技术,分析25例正常早孕、35例早孕自然流产绒毛组织中Igf2基因印迹丢失(loss of imprinting,LOI)情况;采用荧光定量PCR方法检测两种绒毛组织Igf2基因mRNA的表达。结果正常绒毛组织中Igf2基因型为杂合子的样本有11例,其中4例被检测出印迹丢失;自然流产绒毛组织中Igf2基因型为杂合子的样本有16例,其中6例被检测出印迹丢失,二者相比差异无统计学意义(P>0.05)。自然流产绒毛组织中Igf2基因mRNA的表达量为(2.29±0.60)×103copy/μgRNA,明显低于正常绒毛组织中Igf2基因的表达量(3.15±0.88)×103copy/μgRNA,两者比较差异有统计学意义(P<0.001)。结论 Igf2基因的印迹丢失与自然流产无关,Igf2基因mRNA的表达降低与自然流产有关,在早期自然流产发生中可能存在其他机制调节Igf2基因的表达。
Objective To study the imprinting status of insulin like growth factor 2 (Igf2) gene and its role in early spontaneous abortion. Methods From February 2010 to February 2011, 60 pregnant women were treated in the contraceptive department of Shengjing Hospital, China Medical University. PCR and restriction fragment length polymorphism (RFLP) were used to analyze the clinical data of 25 pregnant women Igf2 gene loss in imprinting (LOI) was observed in 35 cases of early pregnancy induced by spontaneous abortion. The mRNA expression of Igf2 in two kinds of villi was detected by fluorescence quantitative PCR. Results Eleven samples with heterozygous Igf2 genotypes were found in normal villi tissues, of which four were found to have loss of imprinting. Sixteen cases of Igf2 genotype heterozygous in spontaneous abortion chorion were detected in 6 of them Lost, the difference between the two was not statistically significant (P> 0.05). The mRNA expression of Igf2 in spontaneous abortion chorion was (2.29 ± 0.60) × 103copy / μgRNA, which was significantly lower than that in normal chorion (3.15 ± 0.88) × 103copy / μgRNA Significance (P <0.001). Conclusion The imprinting loss of Igf2 gene has nothing to do with spontaneous abortion. The decrease of Igf2 mRNA expression is associated with spontaneous abortion. There may be other mechanisms regulating the expression of Igf2 gene in spontaneous abortion.