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Objective: 18F-3deoxy-3-18F-fluorothymidine (18F-FLT) has been described recently as a new tracer for imaging tumor cell proliferation.There are few reports about correlation of 18F-FLT uptake with different situations of cycle cell cycle.Therefore, we conducted the study to observe the uptake of 18F-FLT in vitro of lung adenocarcinoma SPC-A1 cells which were in the different situations of division cycle, and to correlate that relationship with the cellular proliferation of the tumor.Methods: The cell proliferation was blocked by drug intervention.Respectively, the G2/M phase,S phase, G0/G1 phase-based samples were harvested for each batch, a total of three batches of 36 samples.There were about 5 × 106 cells in each culture bottle.1.85×105Bq (5μCi)18F-FLT was added to each bottle and the cells were continued incubating for 60 minutes.Cells were harvested and were divided into two parts.For one parts of cells, detected the synchronization ratio by Flow Cytometry after a 24-hour period, and expressed as percentage in each cycle.For another part of cells, the uptake rate of cells was counted by the γ-counter.Results: 1.The 18F-FLT uptake of every 1 × 106 lung adenocarcinoma SPC-A1 cells was significantly correlated with the cell percentage of S phase cells (r =0.642, P <0.01), and also had a significant correlation with the cell percentage of G2/M phase cells (r =0.695, P =0.001).2.The 18F-FLT uptake of S-phase cells is higher than G2/M phase cells (P<0.05).Conclusions: 18F-FLT uptake had a significant correlation with the the cellular proliferation in vitro lung adenocarcinoma SPC-A1 cells, which has the highest uptake in S phase ceils.It was suggested that 18F-FLT PET/CT imaging maybe a good marker for assessment of response to anticancer drugs.