Influence of Epidermal Growth Factor on Expression of Matrix Metalloproteinase-2 , 9 mRNA in Mouse Embryos

Kim, J H; Chae, H D; Kang, E H; Chu, H S; Jeon, Y P; Kang, B M; Chang, Y S

Abstract

OBJECTIVE
To investigate the influence of epidermal growth factor [EGF] on expression of matrix metalloproteinase-2, 9 [MMP-2, 9] mRNA in mouse embryos Materials and METHOD: From January to June 1998, morula stage mouse embryos were cultured for 48 hours with EGF at concentrations of 0.1, 1.0, 10 and 100 ng/ml. Embryos not treated with EGF were served as control. Reverse transcription-polymerase chain reaction [RT-PCR] has been used to examine the expression of MMP-2, 9 mRNA in developed blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with student t-test and statistical significance was defined as p < 0.05.
RESULTS
The relative quantity [relative volume x intensity] of MMP-2 mRNA expressed in embryos of each group were not differed significantly [control ; 457.7, 0.1 ng/ml group; 525.0, 1 ng/ml group ; 455.0, 10 ng/ml group ; 562.7, and 100 ng/ml group ; 584.5]. There were also no significant differences in the relative quantity of MMP-9 mRNA expressed in embryos among each group [control ; 653.3, 0.1 ng/ml group ; 425.4, 1 ng/ml group ; 360.2, 10 ng/ml group; 394.8, and 100 ng/ml group ; 443.5].
CONCLUSION
This study suggests that EGF itself may not induce the expression of MMP-2 or 9 mRNA in mouse blastocysts.