Tumor cells characteristically exhibit an increased rate of glycolysis. A higher level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was found in human uterine cervical cancers. This study was designed using GAPDH antisense oligodeoxynucleotide (ODN) phosphorothioate (PS) to evaluate how alterations of GAPDH expression in human cervical carcinoma could influence growth inhibition and induction of apoptosis. Northern blot analyses revealed that the levels of GAPDH gene expression were strongly elevated in three cervical carcinoma cell lines (HeLa, CUMC-3, and CUMC-6) compared with normal cervical tissue. Reverse transcription-polymerase chain reaction (RT-PCR) showed that expression of the GAPDH gene was inhibited by 10 microM of GAPDH antisense ODN in all three cell lines. Western blot analysis showed that the levels of GAPDH protein were decreased or absent after GAPDH antisense ODN treatment for 12 days in cultured cervical carcinoma cell lines. Cervical carcinoma cell lines exposed to GAPDH antisense ODN showed reduced cellular proliferation, which was accompanied by reduced colony-forming efficiency. This effect of GAPDH antisense ODN on cultured carcinoma cells was associated with the apoptotic process, including increased DNA fragmentation. These results suggest that future gene therapy using antisense ODN directed against cervical cancer-specific GAPDH mRNA might be another therapeutic tool against human uterine cervical carcinomas.