The Effect of Granulocyte Colony Stimulating Factor and Granulocyte Macrophage Colony Stimulating Factor on Expression of Matrix Metalloproteinase-2, 9 in Mouse Embryos.
- Author:
Chung Hoon KIM
;
Hee Dong CHAE
;
Eun Hee KANG
;
Yong Pil CHEON
;
Byung Moon KANG
;
Yoon Seok CHANG
- Publication Type:Original Article
- Keywords:
Granulocyte colony stimulating factor(G-CSF);
Granulocyte macrophage colony stimulating factor(GM- CSF);
Matrix metalloproteinase-2;
Reverse transcription-polymerase chain reaction(RT-PCR)
- MeSH:
Animals;
Blastocyst;
Colony-Stimulating Factors*;
Digestion;
DNA, Complementary;
Embryonic Structures*;
Granulocyte Colony-Stimulating Factor;
Granulocyte-Macrophage Colony-Stimulating Factor*;
Granulocytes*;
Matrix Metalloproteinase 2*;
Mice*;
Morula;
Reverse Transcription;
RNA, Messenger;
Sensitivity and Specificity;
Sequence Analysis
- From:Korean Journal of Obstetrics and Gynecology
1999;42(10):2205-2213
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
OBJECTIVES: To investigate the effect of granulocyte colony stimulating factor (G-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF) on expression of matrix metalloproteinase-2, 9 (MMP-2, 9) mRNA in mouse embryos. Materials and METHOD: From October 1997 to December 1998, morula stage mouse embryos were cultured for 48 hours with G-CSF and GM-CSF at concentrations of 0.1 pg/ml, 1 pg/ml, 10 pg/ml, 100 pg/ml, 1 ng/ml and 10 ng/ml, respectively. Embryos not treated with G-CSF or GM-CSF 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 Kolmogorov-Smirnov test and analysis of variance (ANOVA). The statistical significance was defined as p< 0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of all G-CSF treatment groups were significantly increased than in the control, especially in 10, 100 pg/ml and 1 ng/ml treatment groups. The relative quantities of MMP-2 mRNA in all GM-CSF treatment groups were also significantly increased than in the control, especially in 100 pg/ml treatment group. The relative quantities of MMP-9 mRNA of all GM-CSF treatment groups except 10 ng/ml group were significantly increased than in the control, especially 10, 100 pg/ml and 1 ng/ml treatment group. However, the relative quantity of MMP-9 mRNA was significantly increased in only 10 ng/ml G-CSF treatment group than in the control and other treatment groups. CONCLUSION: This study suggests that G-CSF and GM-CSF may increase the m-RNA expression of MMP-2 or 9 in mouse blastocysts with the concentration-specific manner.
