Influence of Transforming Growth Factor-alpha on Expression of Matrix metalloproteinase-2 and Matrix metalloproteinase-9 mRNA in Mouse Embryos.

Kim, Chung Hoon; Kim, Sung Hoon; Park, Eun Ju; Hong, Seok Ho; Chae, Hee Dong; Kang, Byung Moon

Original Article

Korean Journal of Obstetrics & Gynecology 2002;45(3):443-449.
Published online March 1, 2002.

Influence of Transforming Growth Factor-alpha on Expression of Matrix metalloproteinase-2 and Matrix metalloproteinase-9 mRNA in Mouse Embryos.

Chung Hoon Kim, Sung Hoon Kim, Eun Ju Park, Seok Ho Hong, Hee Dong Chae, Byung Moon Kang

Department of Obstetrics and Gynecology, College of Medicine, University of Ulsan, Asan Medical Center, Seoul, Korea.

Abstract

OBJECTIVE
To investigate the influence of transforming growth factor-alpha (TGF-alpha) on the expression of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9) mRNA in mouse embryos. MATERIALS AND METHOD: Eight-cell stage mouse embryos were cultured for 48hours with TGF-alpha at concentrations of 1, 10 and 100 ng/ml. Embryos not treated with TGF-alpha served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of MMP-2 and MMP-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 analysis of variance (ANOVA) and statistical significance was defined as p<0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of 10 and 100 ng/m of TGF-alpha treatment groups were significantly increased than in the control and 1 ng/ml of TGF-alpha treatment group (67.2+/-7.5 and 77.4+/-11.6 vs. 38.6+/-4.5 and 43.4+/-6.1, p<0.001). The relative quantities of MMP-9 mRNA of 100 ng/ml TGF-alpha treatment group was significantly increased than in the control and 1 ng/ml TGF-alpha treatment groups (67.6+/-6.5 vs. 36.6+/-14.2 and 40.2+/-11.3, p<0.001, p<0.01, respectively). CONCLUSION: This study suggests that TGF-alpha itself may induce the expression of MMP-2 and 9 mRNA in mouse embryos.

Key Words: Transforming growth factor-alpha (TGF-alpha), Matrix metalloproteinase-2 (MMP-2), Matrix metalloproteinase-9 (MMP-9), Reverse transcription-polymerase chain reaction (RT-PCR)