Abstract: Objective To establish an anoikis-resistant subpopulation of cell lines and to observe the relationship between metastasis and anoikis resistance in esophageal carcinoma cells. Methods Analyze the anoikis-resistant ability of EC9706、KYSE30、YES2 and NEC with Flow cytometry.The cells were cultured in the plates coated with Poly-HEMA to select the anoikis-resistant cells through four rounds.The sub-line NEC-P4 were filtrated from the NEC parent cell.The BALB/c nu Mice were intravenously injected with half million cells per animal for NEC-P4 and examined for metastases development. Results The ability of metastasis of the four cells showed that anoikis-resistant EC9706 and KYSE30 had obvious lung metastasis.Visible lung metastasis of EC9706 was 100% and KYSE30 was 83%.EC9706 had 121metastasis sites per lung and KYSE30 had 83.Anoikis-sensitive YES2 and NEC had only 9 and 7.There were no visible lung metastasis in YES2 and NEC guoup.The anoikis-resistant cells we called NEC-P4 were selected through 4 rounds from the anioikis-NEC parent cells.The apoptosis of NEC-P4 was about 18% and NEC parent was about 78% when cultured in plates coated with poly-HEMA.NEC-P4 group had visible lung metastasis and 5 times of HE metastasis sites per lung in average than NEC group. Conclusion There were close relationship between metastasis and Anoikis-resistance in esophageal carcinoma cells.The anoikis resistant cells selected from parent cells always had high metastasis ability.