Objective: To study the effect of SNC19/ST14 gene transfection and expression into colorectal cancer cells on biological behavior.
Methods: The recombination vector pSecTag2a-SNC19/ST14 was constructed, and transfected into the RKO colorectal cancer cell line by liposome. The transfected cell was screened by real time PCR, Western Blot and Immunohistochemical technique. The population doubling time (T(D)) and cell cycle of the transfected cell were analyzed by MTT assay and flow cytometer. Rhodamine-labeled Phallodin was used to label the cell cytoskeletal protein-F-actin, and the F-actin distribution was observed by confocal scanning microscope. The adhesion ability of the transfected cell to extracellular matrix (ECM) was measured by MTT assay.
Results: The full length Open Reading Frame (ORF) of SNC19/ST14 gene was inserted into the vector pSecTag2a, and transfected into RKO cells, and expressed successfully. The changes of F-actin organization took place in transfected cell. The Adherence ability of the transfected cell to ECM was decreased, but the proliferation ability was not significantly changed except highly expressing of the SNC19/ST14, and not the cell cycle and the apoptosis.
Conclusion: SNC19/ST14 gene was successfully expressed into the RKO cell line, and could influence on the cell cytoskeletal protein (F-actin) organization and on cell adherence ability to ECM, but could not make the cell cycle, apoptosis and proliferation ability change significantly.