Objective：To establish esophageal Ec-109 cell subtypes with high and low migration abilities and to explore the possible mechanisms of their different biological properties. Methods：Ec-109 cell subtypes with high and low migration abilities were selected out by modified Transwell chamber migration experiment. Growth differences of the two cell types were detected by growth curve and doubling time. Migration and invasion abilities of the two cells were observed by Transwell chamber. Expression of RhoC and matrix metal proteinase-9（MMP-9） in the two cells was detected by RT-PCR and Western blot. Results：Ec-109 cell subtypes with high and low migration abilities（Ec-109H and Ec-109L） were successfully selected out by modified Transwell chamber migration experiment. No statistical difference was observed between Ec-109H and Ec-109L in growth rate in vitro or doubling time（P=0.105）. Num－bers of migrated cells were 124.9±6.8 and 43.9±6.3 respectively（P=0.000）. Numbers of invaded cells were 50.0±9.7 and 15.2±5.1 respectively（P=0.000）. Expression of RhoC was higher in Ec-109H than in Ec-109L demonstrated by RT-PCR and Western blot（P=0.000），but there was no statistical difference in expression of MMP-9 between the two cell types（P=0.523 and P=0.655）. Conclusion：RhoC plays an important role in migration and metastasis of esophageal cancer，while MMP-9 does not.