Transfer of p14~(ARF) gene in human tumor cell lines inhibits cell proliferation

Objective: To clone p14 ARF gene from cells of the human normal liver cell line L02 and explore its inhibitory effect on tumor cell proliferation. Methods: p14 ARF cDNA was cloned from L02 cells by using RT-PCR method and inserted into the BamHI and EcoRI sites of pcDNA3 (called pcDNA3ARF). Lipofectamine mediated pcDNA3ARF was transferred into tumor cell lines, such as Hep2, KB, KB-v200, HepG2, HLE, Huh7, MCF7 and MCF7/ADR. The number of colonies of these tumor cell lines were calculated after more than two weeks' screening in DMEM medium containing G418. Results:The colony-forming efficiency of the p14ARF -transfected KB-v200 and MCF7/ADR drug-resistant cells was 13±2.0 and 21±1.5 , respectively, compared with 86±3.1(15%)and 126±7.5(17%)in the cells transfected with empty vector pcDNA3, and in the p53 positive tumor cell lines, such as HepG2(47%), Hep2(57%), MCF7(58%) cells, the inhibition was also found,whereas in the p53 negative tumor cell lines, such as KB(74%), HLE(65%), Huh-7(80%) cells, the inhibitory effect was milder. Conclusions: p14 ARF can suppress the growth of the p53 positive tumor cell lines in a p53-relevant manner, while in the p53 negative, drug-resistant tumor cells, p14 ARF has strong inhibitory effect, indicating the inhibitory effect of p14 ARF on growth of drug-resistant tumor cells is independent on p53, at least in KB-v200 and MCF7/ADR drug-resistant cells.