Oct4(Octamer-binding transcription factor4,Oct4)基因是多潜能干细胞的标志之一,为了解Oct4基因在脐带、脐血、羊水中的表达情况,本研究以人胚胎干细胞为阳性对照,β-actin基因为内参基因,利用半定量RT-PCR(Reversetranscrible polymerase chain reaction,RT—PCR)对上述样品检测(各20份),并对扩增片段进行测序和同源分析。结果显示,源于上述样品的RT—PCR扩增片段与人Oct4基因eDNA序列同源性为98%,Oct4基因在脐带、脐血、孕中期羊水和孕晚期羊水中的阳性表达率分别为95%(19/20)、80%(16/20)、90%(18/20)和80%(16/20);在脐带、中期羊水和晚期羊水中,Oct4基因的转录水平没有显著性差异(P〉0.05),但显著高于脐血(P〈0.05)。结果表明Oct4基因在脐带、脐血和羊水中表达,但不同样品有差异。
Oct4 (Octamer-binding transcription factor 4, Oct4) was one of the markers used to evaluate pluripotency or undifferentiated status of stem cells. To investigate expression of the gene Oct4 in umbilical cord, umbilical cord blood and amniotic fluid from human, we analysed 20 samples from women of pregnancy stage or full-term delivery by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), respectively, and PCR products were sequenced and compared with the cDNA sequence of human-derived Oct4 published in GenBank to check specificity of PCR reaction. The RT-PCR results showed that homolog degree of RT-PCR amplified products with the cDNA sequence of human-derived Oct4 achieved to 98%, and Oct4 expressed positively in the samples of 95% (19/20) umbilical cord, 80% (16/20) umbilical cord blood, 90% (18/20) second-trimester amniotic fluid and 80% (16/20) third-trimester amniotic fluid. Expres- sion levels of Oct4 exhibited no statistically significant differences in the umbilical cord, second- and third-trimester amniotic fluid (P〈0.05), while significantly higher than in umbilical cord blood. These results suggested that Oct4 expressed in umbilical cord, umbilical blood and amniotic fluid from human, but the difference existed in different samples.