Influence of Hepatocyte Growth Factor on its Receptor (c-met) expression and Invasion in HT Cell Line. |
Jong Chul Shin, Jee Hyun Lee, Dong Eun Yang, Min Jeong Oh, Hee Bong Moon, Young Lee, Sa Jin Kim, Chong Seung Yi, Soo Pyung Kim |
Department of Obstetrics and Gynecology, Catholic University Medical College, Seoul, Korea. |
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Abstract |
OBJECTIVE To evaluate the invasive capacity of trophoblastic cells when treated with HGF, and see whether HGF induces c-met expression in vitro. METHODS: HT cell line was treated with recombinant HGF (rHGF) at different concentrations (0, 10, 50 and 100 ng/mL) and cultured for 24 hours. To evaluate according different time of culture, HT cell line was pre-treated with 50 ng/mL rHGF and cultured for 24, 36, 48 and 72 hours. We analyzed the expression of c-met mRNA using reverse transcriptase-polymerase chain reaction and the expression of c-met protein using western blot in each samples. We also observed cellular invasion capacity through the invasion assay under a microscope and confirmed 72 kDa gelatinase and 92 kDa gelatinase expression patterns by zymography assay. RESULTS: The expressions of c-met mRNA and protein were increased in all concentrations of rHGF, compared with that of normal control although it was not in dose-dependent fashion. In invasion assay, the number of invaded HT cells were increased in dose-dependent fashion, compared with that of normal control. In zymography ssay, the expression of 72 kDa gelatinase was increased in dose-dependent fashion, compared with the control. However, 92 kDa eglatinase was not detected in any studied group. CONCLUSION: These results suggests that HGF might be related to upregulation of trophoblast cell invasiveness by activation of c-met and subsequent induction of 72 kDa gelatinase. |
Key Words:
Trophoblast, HGF, c-met, Invasiveness |
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