Influence of Hypoxia on Vascular Endothelial Growth Factor and Basic Fibroblast Growth Factor in Cultured Human Trophoblast / 대한산부인과학회잡지

OBJECTIVE: To investigate whether the hypoxic condition influences on the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA in the cultured human trophoblast. METHODS: Trophoblasts were isolated from the normal placenta in early pregnancy (6-10 weeks in gestation). Isolated trophoblasts were cultured under normoxic (5% CO2, 95% humid air in incubator) and hypoxic (MERCK, 1% O2, 99% CO2) conditions for 24, 48 and 72 hours, respectively. Total RNA was extracted from the cultured trophoblasts in each culture condition. The expressions of VEGF and bFGF mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis. RESULTS: The expression of VEGF189 mRNA was significantly increased in the hypoxic condition compared to the normoxic condition and control after 24 hours (p<0.05, p<0.05, respectively). The expression of VEGF206 mRNA was also significantly increased in the hypoxic condition compared to the normoxic condition and control after 48 hours (p<0.05, p<0.05, respectively). However, there was no significant difference between the normoxic and hypoxic conditions in the expression of VEGF121 and VEGF165 mRNA. The expression of bFGF mRNA was significantly increased in the hypoxic condition compared to the normoxic condition and control at 24 hours and 48 hours (p<0.05, p<0.05, respectively). bFGF mRNA was more expressed than VEGF mRNA in the hypoxic condition. CONCLUSION: These findings suggest that the hypoxic condition may stimulates the expression of bFGF and VEGF mRNA, and besides bFGF may be a more potent inducer of angiogenesis rather than VEGF in early human gestation.

Influence of Hypoxic Condition on Invasion of Cultured Trophoblast / 대한주산의학회잡지

OBJECTIVE: To investigated whether lowering oxygen tension affects invasion of cultured trophoblast. METHODS: Trophoblasts were isolated from the normal placenta in early pregnancy(6-10 weeks in gestation). Isolated trophoblasts were cultured under normoxic(5% CO2, 95% humid air in incubator) and hypoxic(MERCK, 1% O2, 99% CO2) conditions for 24, 48 and 72 hours, respectively. The proliferation ability was measured using [H3] thymidine assay. Total RNA was extracted from the cultured trophoblasts. The expressions of matrix metalloproteinase(MMP-2) and tissue inhibitor of metallo- proteinase(TIMP-2) were determined by reverse transcription- polymerase chain reaction(RT-PCR) and Northern blot analysis. The invasiveness of cultured trophoblast was observed using in vitro invasion assay. RESULTS: [H] thymidine assay indicated that cellular DNA synthesis was not affected by the culture condition. The expression of MMP-2 mRNA was decreased at 24 hours and then progressively increased in the time-dependent manner in each culture condition. The expression of TIMP-2 was decreased in the time-dependent manner under hypoxic condition. In vitro invasion assay revealed that the cultured trophoblasts under hypoxic condition has more invasive ability than them under normoxic condition. CONCLUSION: These data suggests that hypoxic condition may stimulates the invasion of trophoblast in the human placentation. And MMP-2 and TIMP-2 may be related to control their invasiveness under hypoxic condition.