In order to study the effect of microfilament cytoskeleton on SHEEC esophageal cancer cells and the biological activity variation of cancer cells caused by antisense blocking of NGAL gene expression, making use of antisense expressing vector of different length NGAL gene segments and antisense oligonucleotide segments of thio-modification to transfer the SHEEC cancer cells of esophagus, a series of subcellular clones aiming at the blocking of NGAL gene expression of SHEEC esophageal cancer cells were established by means of G418 selection. On the basis of the fluorescent dual-labeling of F-actin and DNA within cells and after antisense blocking of NGAL gene expression, the content of F-actin and DNA, the F-actin cytoskeleton structure in SHEEC cell and the variational characteristics of biological action on the tumor cell were tested by use of flow cytometry and confocal scanning laser microscope (CSLM). The results showed that F-actin level in the SHEEC cells was obviously reduced after antisense blocking and it was similar to that in SHEE cells, but no obvious change of cellular division index was found. This indicates that the blocking of NGAL gene expression has an obvious effect on the microfilament cytoskeleton of SHEEC cells, but no obvious influence on SHEEC cell proliferation. With the technique of CSLM, it is revealed that the blocking of NGAL gene expression could make the cellular F-actin cytoskeleton of SHEEC cell uniformly distributed, F-actin body evidently decreased, the joint structure of the cell reestablished, the cellular structure more compact, and the main structure characteristics of SHEEC cell and those of F-actin cytoskeleton structures of SHEE cell consistently approached. The result suggests that SHEEC cellular F-actin cytoskeleton of the esophagus cancer could be obviously affected by antisense blocking of NGAL gene expression, and it is conjectured that microfilament cytoskeleton F-actin in cancer cell could be a function key of NGAL gene that plays a role in SHEEC cancer cell.