| Role of nitric oxide and interleukin-1beta on expression of matrix metalloproteinase-2 in TL cell line. |
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Jae Dong Lee, Jong Chul Shin, Dong Eun Yang, Hee Bong Moon, Jee Hyun Lee, Hyun Young Ahn, Gui SeRa Lee, Sa Jin Kim, Su Pyung Kim |
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Department of Obstetrics and Gynecology, College of Medicine, Catholic University, Seoul, Korea. |
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| Abstract |
OBJECTIVE
We studied to investigate whether nitric oxide (NO) and IL-1beta modulate MMP-2 and MMP-9 using TL cell line obtained from the normal term placenta. METHODS: After culturing TL cell line for 4 hours, we treated 0.1 mM of SNAP (NO donor) and 50 ng/ml of IL-1beta for 0, 1, 3, 6, and 12 hours, for investigating changes from time. We treated SNAP of 0, 0.01, 0.1, and 0.5 mM for 12 hours and IL-1beta of 0, 1, 10, and 50 ng/ml, for investigating changes from concentration. After extraction of total RNA, we performed reverse transcriptase-polymerase chain reaction (RT-PCR), gelatine zymography and Western blot analysis, for investigating expression of MMP-2 and MMP-9. RESULTS: MMP-9 was not observed in TL cell line. The expressions of MMP-2 mRNA and protein were gradually increased according to the culture time in SNAP treated group. The expressions of MMP-2 mRNA and protein were gradually increased according to the culture time in IL-1beta treated group. The expression of MMP-2 protein was not more increase in SNAP/IL-1beta-treated group than in IL-1beta treated group. The expression of MMP-2 protein was more reduced in SNAP/hemoglobin treated group than in SNAP treated group. MMP-2 protein activity was only increase in SNAP treated group. CONCLUSION: These results indicate that NO, rather than IL-1beta, upregulates the MMP-2 in TL cell line and furthermore may influence in the invasive process of trophoblasts. |
| Key Words:
Trophoblast, Matrix metalloproteinase-2, Nitric oxide, Inteleukin-1beta |
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