ABSTRACT
Objective To study Edg4 and Edg7 expression in placenta of women with hypertensive disorder complicating pregnancy,and to investigate the relation between the expression of lysophosphatidic acid(LPA)and hypertensive disorder complicating pregnancy.Methods Immunohistochemical SP method was used to measure the expressions of Edg4 and Edg7 in placenta of women with normal pregnancy,20 women with gestational hypertension,20 with mild preeclampsia,and with severe preeclampsia.Results (1)Location:immunohistoehemical staining for Edg4 and Edg7 protein were located at the membrane and endoehylema of cytotrophoblast as well as decidua cells.(2)The positive expression of Edg4 protein and Edg7 protein on membrane and endochylema of cytotrophoblast was 25% and 20%(normal women),60% and 40%(gestational hypertension),80% and 65%(mild preeclampsia),and 83.3% and 86.7%(severe preeclampsia).The expression of Edg4 and Edg7 protein in mild preeclampsia and severe preeclampsia was significantly correlated with the degree of differentiation(P0.05). (3)The positive expression of Edg4 protein and Edg7 protein on membrane and endochylema of decidua was 20% and 25%(normal pregnancy),55% and 50%(gestational hypertension),70% and 55%(mild preeclampsia),and 83.3% and 73.3%(severe preeclampsia)respectively.The expression of Edg4 and Edg7 protein in mild preeclampsia and severe preeclampsia showed a significant correlation with the degree of differentiation(P0.05).Conclusions The high expression of Edg4 and Edg7 protein in the placentas of patients with hypertensive disorder complicating pregnancy indicates that LPA combines with Edg4 and Edg7,inducing the occurrence of hypertensive disorder complicating pregnancy.
ABSTRACT
Objective To construct the recombinant eukaryotic expression vector pRNAT-U6,1- siEdg4 which curries small interfering RNA(siRNA)of Edg4 and observe the silencing effect of Edg4 gene targeted siRNA in ovarian cancer cell line SKOV3.Methods The Edg4 gene-targeted hairpin siRNA sequence was designed according to the Edg4 sequence in Genbank,and the two complementary oligo nucleotide strands were synthesized and annealed and inserted into the pRNAT-U6.1 plasmid to build a recombinant Edg4 siRNA eukaryotic expression vector,which was sequenced and identified to contain the correct Edg4 siRNA sequence.The human ovarian carcinoma cell lines SKOV3 were transfeeted with the vector using lipofeetamine method.The efficiency of transfecting cells was observed with fluorescent microscope and the mRNA expression level of Edg4 gene was detected by real time quantitative PCR.The LPA levels in cell supernatants were detected using a biochemical method.And the apoptosis of SKOV3 cells induced by the vector was evaluated by flow cytometry.Results The recombinant eukaryotic expression vector was confirmed to contain correct Edg4 siRNA sequence by PCR and sequencing.After transfection large amounts of green fluorescence were seen in plasma and nuclei of SKOV3 cells and the positive cell rates were 64%.The expression level of Edg4 mRNA in transfeeted SKOV3 cell line was significantly decreased (0.05?0.01 vs 0.29?0.04,P