| Objective: To investigate the effect of the ligand of peroxisome proliferator-activated receptor Rosiglitazone using alone and combinated with Cisplatin on human lung adenocarcinoma A549 cells'growth and apoptosis in vitroto, and to explore its mechanism.Methods: human lung adenocarcinoma A549 cells was cultures in vitro. A549 cells were treated with different concentrations of ROZ or DDP or both using MTT (tetrzolium-based colorimetric assay, MTT)measurement of different concentrations of ROZ in human lung adenocarcinoma A549 cell proliferation inhibition, choose suitable dose. Distribution of cell cycle and apoptosis were measured by flow cytometry and The expression of PPARγ,NF-κB,survivin, caspase-3 protein was studied by immunohistochemistry.Results:1 MTT assay indicated that ROZ whose concentration excessed 50μmol/L could inhibit the proliferation of A549 cells. Different concentration of ROZ treat the human lung adenocarcinoma A549 cells in 24~72h, the OD value of experimental group were decreased to varying degrees, compared with the control group, there was a significant difference (P<0.01). With the increase in ROZ concentration and the role of the extended time, the OD values gradually decreased. Different concentration of ROZ treat the human lung adenocarcinoma A549 cells in 24~72h, the inhibitory rate were decreased from 9.94%~48.2% to 25.6%~91.7%, 1.25μmol/L of ROZ on A549 cells,no significant toxicity (IR<10%), but if Combined with DDP it could enhances DDP on A549 cell proliferation inhibition (q>1.15).2 Distribution of cell cycle was measured by flow cytometry, the results were: 50μmol/L ROZ treat A549 cells after 48h there apoptosis rate was 7.43±0.01%. With a disability, and 100μmol/L ROZ treat A549 cells after 48h the apoptosis rate of A549 cells increased to 11.32±0.10%. With the increase of ROZ concentration, G1 phase cells gradually increased, S phase cells decreased. ROZ can be prompted A549 cell cycle arrest in G1/G0 phase, the effect has a dose-dependent manner; 48 hours of natural apoptosis of A549 cells was 1.72±0.038%, 1.25μmol/L ROZ separate treat the cells in 48 hours the A549, apoptosis rate was 1.84±0.794%. It is suggesting that 1.25μmol/L ROZ does not induce the apoptosis; different concentrations of DDP (1.0,2.0,4.0mg/L) with A549 cells for 48 hours, the A549 cells can be induced apoptosis, and combinated with the ROZ (1.25μmol/L) the apoptosis rate increased, DDP will enable the A549 cells in G0/G1 phase cells increased, S phase cells decreased in a dose-dependent manner. ROZ (1.25μmol/L) and United Group DDP and DDP cell cycle no significant changes in group.3 Immunohistochemical method results showed that: the normal A549 cells expressed PPARγ, NF-κB, Survivin, Caspase-3 protein, adding different concentrations of ROZ for 48 hours, in the low and high dose group PPARγprotein IHS values were:3.60±0.75 and 8.40±0.75, as well as the negative control group was 1.40±0.24. There was a significant difference (P <0.05) between each dose group and negative control group. In the low and high dose of ROZ groups of cancer cells NF-κB protein in the IHS values were: 4.80±0.49,2.40±0.40 as well as the negative control group was 8.40±0.60. There was a significant difference (P<0.01). In the low and high dose of ROZ groups of cancer cells Survivin protein in the IHS values were: 6.80±0.80, 2.80±0.49, as well as the negative control group was 9.60±0.75. There is significant difference (P<0.01). In the low and high dose of ROZ groups of cancer cells Caspase-3 protein IHS values were: 3.20±0.49, 6.00±0.63, as well as the negative control group was 1.60±0.24. There is also a significant difference compared with the control group. NF-κB, Survivin protein expression with the role of the increase in concentration decreased, while the PPARγ, Caspase-3 protein expression with the role of the concentration of ROZ increases the expression and the location changes.Conclusion:1 ROZ within a certain dose range can inhibit the growth of lung adenocarcinoma A549 cells and induced the apoptosis, block cells in G1 phase, the role was a certain amount of time, dose-dependent manner.2 ROZ can enhance cisplatin on A549 cell growth inhibition and induction of apoptosis.3 ROZ inhibition of A549 cell proliferation and sensitizing effect of chemotherapy may be associated with activation of PPARγ, reduced NF-κB, Survivin protein and increase of Caspase-3 protein expression.
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