Effect Of Proliferation And AEG-1Expression Of Human Lung Cancer Cell Line A549Induced By NF-κB Inhibitor Pyrrolidine Dithiocarbamate

ObjectiveTo explore the effect of pyrrolidine dithiocarbamate (PDTC) on proliferation of human lung cancer cell line A549and expression of astrocyte elevated gene-1(AEG-1), in order to evaluate PDTC as a potential drug for chemoprevention and chemotherapy of human lung cancer.Methods1. A549cells cultured in vitro were treated with PDTC of different concentrations (25、50、100、200μmol/L) for24h,48h and72h respectively. MTT assay detected the inhibition rates of cell growth.2. Flow cytometry (FCM) analyzed the changes of cell cycle after treatment by drugs.3. Hoechst33342observed the apoptosis of cells after treatment by drugs with the fluorescence microscope.4. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot detected the expression of the AEG-1gene and protein before and after treated with drugs.5. SPSS17.0statistical software was applied for all the data statistics.x±s and one-way ANOVA or Kruskal-Walli Test was used to compare the quantitative data, and LSD Test was used to multiple comparisons. P<0.05was consider significant.Results1. PDTC inhibited the A549cells proliferation in a time-dose dependent manner. After treated with PDTC for24hours,48hours and72hours, the inhibition rate of cell proliferation of all the concentration groups are higher compare to the control group, and the differences were statistically significant (P<0.05)2. PDTC can change the distribution of cell cycle. In contrast to the control, the percentage of cells in G0/G1phase increased significantly in PDTC group. The distribution of the four concentration groups are different compare to the control group, and the differences were statistically significant (P<0.05), in a time-dose dependent manner.3. After the fluorescence staining, the results observed by converted fluorescence microscope showed that there are irregular shaped cells treated by drugs, and with the higher concentration of drug, there were apparent apoptotic morphological changes in the cells.4. The mRNAand protein expression of AEG-1in A549cells which induced with increasing concentration of PDTC were obviously lower than those in normal cells, and the differences were statistically significant(P<0.05), while the inhibitory effects presents in does-dependent manner.ConclusionsPDTC can inhibit the proliferation of A549cells, change the distribution of cell cycle and also can induce the apoptosis of A549cells in a time-dose dependent fashion. The mechanism may be related to the down-regulated expressions of NF-kB and AEG-1genes induced by PDTC.