In an attampt to establish the functional expression of STGC3 with doxycycline (Dox) induced Tet-on regulating system in human nasopharyngeal carcinoma cell line CNE2, an ideal experimental platform was provided for further studies of STGC3. pTet-on regulating plasmid was transfected into CNE2，and stable expression of Tet-on was established in CNE2 through G418 select. Then the response plasmid of recombinant pTRE-STGC3 was steadily transfected into positive CNE2/Tet-on cells with hygromycin screen. Dox was used to induce the expression of STGC3 and a cell clone sensitive to Dox was selected. The best-induced concentration was determined with different concentration of Dox induction. Growth curves, clone formation rate and cell cycle distribution were detected after STGC3 gene up-regulated expression with Dox induction. The growth capacity and clone formation potential of CNE2/Tet /pTRE-STGC3 was significantly suppressed, compared with the controls (P＜0.05). FCM analysis indicated that G0/G1 phrase cell rate of CNE2/Tet /pTRE-STGC3 was markedly higher than that of the controls and CNE2/Tet/pTRE-STGC3 cells were arrested in G0/G1 phase of cell cycle. Functional expression of STGC3 under Dox induced Tet-on regulation system was successfully established in CNE2, which provided an ideal experimental platform for further functional study of STGC3.