The Expression Of Glypican-3 And Glypican-6 In Macrosomic Placenta

Macrosomia is referred to newborn infants with birth weight greater than 4000g. With the increase of life standard and the prevalence of knowledge about pregnancy-related health care in China, many pregnant women eat too much and exercise too less, which resulted in the rise of incidence of macrosomia. Although the majority of macrosomia is physiologically healthy infants, some macrosomia infants got some symptoms. In clinic, pregnancy combined with diabetes, pregnant obesity, genetic factors, and endocrine, etc, are the common sinner of macrosima. But, the etiologic definition of macrosomic is still dependent on the biomedical and molecular methods. In recent years, the relationship of hormone and macrosomic development gained more and more attentions. A great number of studies have shown that insulin, IGFs, thyroxin, growth hormone, leptin and FGF22, etc, are correlated with fetal growth. Among them, IGF is certainly one of the most important macrosoma-related factors, any changes of expression or imprinted status of which could led to fetal overgrowth and anomaly.Glypicans are a family of heparan sulfate proteoglycans (HSPGs) that are linked to the exocytoplasmic surface of the plasma membrane by a lycosyl-phosphatidylinositol (GPI) anchor. Through the interaction with growth factors, ECM-related factors, cell-adhesion molecules and degradation pathway-related molecules, glypicans regulate cell proliferation and differentiation. A couple of evidences have indicated that glypicans are related with growth and development. In glypican superfamily, glypican-3 is the most important regulator of growth. The relationship of glypican-3 and Simpson-Golabi-Behmel has been reported. But, as the new member of glypican family, the expression of glypican-6 has been found in human placenta, but the relationship of glypican-6 and fetal growth is still unknown.Objective: our study will investigate the expression difference of glypican-3 and glypican-6 between macoromic and normal placenta, the result of which will be beneficial to the interpretation of glypican's role in macrosomic development.Methods:1.Collection of 24 normal newborn infant placenta and 17 macrosomic placenta, and conservation of them in iquid nitrogen.2.After extraction of total RNA of placenta tissues using TRIzol kit, SuperScriptTM First-Strand Synthesis System is used for reverse transcription. Then, glypican-3, glypican-6 was amplified using Takara PCR kit, with house-keeping gene GAPDH as internal control.3.Total protein of placenta tissues were prepared by using RIPA, then bradford protein assay was performed to quantify the protein concentration. After SDS-PAGE separation, proteins was transferred to PVDF membrane using semi-dry methods. Then, after incubation with the primary antibodies of glypican-3, glypican-6, GAPDH ( as internal control), exposure was performed by using ECL kit.4.Quantity One was used to quantify the gray density of bands, the result of which was statistically analyzed with SPSS15.0 software. P<0.05 is deemed as significant difference.Results: The expression of glypican-3 of macrosomic placenta tissues in mRNA or protein level are lower than that of normal placenta, with statistical significance (p<0.05). The expression of glypican-6 in the two groups has not significant difference (p>0.05).Conclusions:The expression of glypican-3 in macrosomic placenta is lower than that of normal placenta, which suggested an important role of glypican-3 in macrosomic development.