transforming-growth-factor-beta and Hydatidiform-Mole

Topics: Adolescent; Adult; Animals; Chromosome Mapping; Diptera; Feedback; Female; Follicle Stimulating Hormone; Humans; Hydatidiform Mole; Inhibins; Luteinizing Hormone; Male; Middle Aged; Ovary; Pregnancy; Transforming Growth Factor beta; Xenopus

Previous studies indicate that epigenetic modifications play an important role in transcriptional regulation and contribute to the pathogenesis of gestational trophoblastic disease, including complete hydatidiform moles (CHMs). However, the underlying mechanisms and the critical genes have not been clearly identified. In the present study, we developed a novel technique, NotI subtraction and methylation-specific genome subtractive hybridization (MS-G-SH), as a method of screening for methylation changes between hydatidiform moles and villi. Following NotI subtraction and hybridization, three different positive DNA clones were found in 110 random clones of DNA samples. Most importantly, two DNA clones having long CpG islands and high homology with exons of insulin-like growth factor 2 (IGF2) and transforming growth factor-β (TGF-β) were identified using bioinformatic tools. After bisulfite treatment and methylation-specific PCR, the specific methylation of certain exons of IGF2 and TGF-β was identified. In addition, the mRNA expression levels of these two genes were markedly different. In conclusion, this novel MS-G-SH technique is an alternative and effective approach for the detection of specific DNA methylation.

Topics: Base Sequence; Chorionic Villi; Computational Biology; CpG Islands; DNA Methylation; Epigenesis, Genetic; Epigenomics; Female; Gene Expression Regulation; Genetic Loci; Genome, Human; Humans; Hydatidiform Mole; Insulin-Like Growth Factor II; Molecular Sequence Data; Pregnancy; Sequence Alignment; Transforming Growth Factor beta; Trophoblasts

The semiquantitative reverse transcription polymerase chain reaction was employed to detect the expression of transforming growth factor beta (TGF-beta) and insulin-like growth factor (IGF) in complete hydatidiform mole, normal first-trimester villi, the normal term placenta (after vaginal/abdominal deliver) and the preeclamptic placenta at term. The expression of IGF-I mRNA was seen in all five tissues, but its level was much lower in the term placental tissues with preeclampsia than in other tissues. The content of IGF-I mRNA in villous tissues from molar pregnancy was slightly higher than in normal first-trimester villi. IGF-II mRNA was detected at similar levels in all three sorts of term placental tissues. However, the expression level of IGF-II mRNA in tissues of complete molar pregnancy was significantly lower than in normal first-trimester villi. TGF-beta(3) was found expressed in all five tissues, while TGF-beta(1) and TGF-beta(2) mRNA were not detected. Compared to the normal first-trimester villi, the expression of TGF-beta(3) in complete hydatidiform molar tissues was comparatively higher. Furthermore, the expression levels of TGF-beta(3) in the preeclamptic placenta and the normal placenta after cesarean birth were higher than in the placenta after vaginal delivery. We concluded that, the change of TGF-beta and IGF expression in placental tissues might be involved in the development of trophoblastic diseases of pregnancy.

Topics: Adult; Chorionic Villi; Electrophoresis, Agar Gel; Female; Humans; Hydatidiform Mole; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Placenta; Pre-Eclampsia; Pregnancy; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta