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Objective To investigate the effect of hepatitis B virus X (HBx)protein on the apoptosis of placental trophoblastic cells and its potential mechanism.Methods A pcDNA3.1 expression vector of HBx gene was constructed and transfected into JEG-3 and HTR-8 human placental trophoblastic cell lines,respectively.After transfection for 48 h,RT-PCR and immunofluorescence analyses were made to detect HBx mRNA and protein expressions.Flow cytometry was used to detect the early apoptosis status of JEG-3 and HTR-8 cells.The expressions of PI3K and p-Akt were detected by immunofluorescence and Western blotting.Results After transfection for 48 h,RT-PCR and immunofluorescence analyses showed that HBx mRNA and protein expressions were detected in JEG-3 and HTR-8 cells.Flow cytometry revealed that early apoptosis of JEG-3 and HTR-8 cells was reduced by pcDNA-HBx transfection (P <0.05).Immunofluorescence and Western blotting showed that PI3K and p-Akt were significantly upregulated in HTR-8 cells (P < 0.05 ).Conclusion HBx gene can be transfected into JEG-3 and HTR-8 human placental trophoblastic cell lines,respectively.After the transfection,the early apoptosis of JEG-3 and HTR-8 cells is reduced.Its inhibition on apoptosis is related to the activation of the PI3K/Akt signaling path-way.
RESUMO
Objective To investigate the effect of hepatitis B virus X (HBx)protein on the apoptosis of placental trophoblastic cells and its potential mechanism.Methods A pcDNA3.1 expression vector of HBx gene was constructed and transfected into JEG-3 and HTR-8 human placental trophoblastic cell lines,respectively.After transfection for 48 h,RT-PCR and immunofluorescence analyses were made to detect HBx mRNA and protein expressions.Flow cytometry was used to detect the early apoptosis status of JEG-3 and HTR-8 cells.The expressions of PI3K and p-Akt were detected by immunofluorescence and Western blotting.Results After transfection for 48 h,RT-PCR and immunofluorescence analyses showed that HBx mRNA and protein expressions were detected in JEG-3 and HTR-8 cells.Flow cytometry revealed that early apoptosis of JEG-3 and HTR-8 cells was reduced by pcDNA-HBx transfection (P <0.05).Immunofluorescence and Western blotting showed that PI3K and p-Akt were significantly upregulated in HTR-8 cells (P < 0.05 ).Conclusion HBx gene can be transfected into JEG-3 and HTR-8 human placental trophoblastic cell lines,respectively.After the transfection,the early apoptosis of JEG-3 and HTR-8 cells is reduced.Its inhibition on apoptosis is related to the activation of the PI3K/Akt signaling path-way.
RESUMO
Objective To assess the association between the depth of trophoblastic penetration into the tubal wall with serum concentrations of vascular endothelial growth factor (VEGF) and β-human chorionic gonadotropin (β-HCG). Meth-ods Eighty patients with a diagnosis of tubal pregnancy in the ampullary region underwent radical surgical treatment (sal-pingectomy), were included in this study. The serum levels of VEGF andβ-HCG were detected on the day of surgery. The se-rum level of VEGF was measured by ELISA. The serum level ofβ-HCG was quantified with a two-site immunofluorimetric assay based on the direct sandwichtechnique. Histological material was stained with Masson's trichrome to identify muscular fibers. Immunohistochemical staining was used for human placental lactogen (hPL) to identify intermediate trophoblast and determine the depth of trophoblastic invasion into the tubal wall. The ampullary pregnancies were classified histologically ac-cording to the depth of trophoblastic infiltration into the tubal wall. Results The mean serum values of VEGF andβ-HCG were significantly lower in patients with stage I tubal infiltration than those of stageⅡ, and which were significantly lower in patients with stageⅡthan those in stageⅢ(P<0.05). The threshold serum value of VEGF was 308.6 ng/L, the sensitivity was 100.0%and the specificity was 92.6%for stageⅠand stageⅡ. The threshold serum value of VEGF was 431.9 ng/L, the sensitivity was 79.3%and specificity was 79.2%for stageⅡandⅢ. The threshold serum value ofβ-hCG was 2 509.6 IU/L, the sensitivity was 91.7%and specificity was 81.5%for stageⅠand stageⅡ, and levels of 13 142.6 IU/L, 72.4%and 95.8%for stageⅡand stageⅢ. Conclusion The depth of trophoblastic penetration into the tubal wall is associated with maternal serum concentrations of VEGF andβ-HCG, which can be used as the evaluation index for histological staging of trophoblas-tic tissue infiltration.