Effect Of PPAR Alpha Agonist Fenofibrate On The Growth And Metastasis Of Lung Cancer And Relevant Mechanism

Objective: To observe the effects of Peroxisome Proliferator—activated receptor α agonists fenofibrate on the influence of cell growth and metastasis in A549 cells line,discuss PPARα, MMPs and TIMPs meaning in the fenofibrate anti-tumor effect.Methods :Lung adenocarcinoma cell line A549 cultured in vitro were divided into six drug groups(0、12.5、25、50、75、100μmol/L),A549 cells were exposed to fenofibrate, MTT was employed to investigate proliferation,flow was used to detect the cell cycle and apoptosis of A549 cells,Hoechest staining to observe the morphological changes of A549 cells, wound healing test and Transwell chamber were used to detect the effect of drug intervention on the migration and metastatic ability of A549 cells, total RNA and proteins was extracted from different drug concentrations of A549 cells, RT-PCR test PPARα, MMP-2, MMP-9 and TIMP-1 and TIMP-2 m RNA expression,and Wertern blot the expression of MMP-2 and MMP-9 protein levels, enzyme spectrum method to detect MMP-2 and MMP-9 enzyme activity in cell cultures.Results:1. Cell proliferation inhibition rate: compared with control group, with the increasing of fenofibrate drug concentration and action time the growth of A549 proliferation inhibition rate significantly increased. 2.Apoptosis detection: fenofibrate can induce the apoptosis of A549 lung cancer cell, high concentration group effect more apparent. 3.Cell cycle detection: fenofibrate can make A549 lung cancer cell block in G2/M phase and inhibition the cell proliferation. 4.AS shown by Hoechest 33258 staining, under the action of fenofibrate A549 shrinkage, part of the cell membranes by pyrolysis nucleus pycnosis, a thick dense, and high concentration group accompanied the formation of apoptotic bodies. 5.Wound healing test show that: 1. After 48 hours, the negative control group cell scratch basic healing, with the increase of concentration of fenofibrate, the greater the scratch width,each cell migration rates: control group:( 88.21±0.09) %,12.5μmol/L group:(71.70±0.47)%, 25μmol/L group:(48.23±0.09)%,50μmol/L group:(30.45±0.53)%,75μmol/L group:(20.90±0.47)%, 100μmol/L group:(16.20±0.29)%,(p<0.05).6. Transwell chamber showed that :compared with negative control group,with the increase of concentration of fenofibrate, the number of cells through the basement membrane gradually reduced, The highest concentration of fenofibrate through the chamber under the action of basement membrane cells less. Each invasive inhibition rates: 12.5μmol/L group:(23.10±0.62)%, 25μmol/L group:(30.97±0.45)%, 50μmol/L group:(40.35±0.19)%, 75μmol/L group:(48.50± 0.23)%, 100μmol/L group:(65.78±0.58)%,(p<0.05). 7. RT-PCR results showed that: compared with control group, fenofibrate treatment after 48 hours, with the increase of concentration of fenofibrate, PPARα,Timp-1 and Timp-2 m RNA expression increased, and the expression of MMP-2 and MMP-9 m RNA decreased.8.Wertern blot experiments show that: with fenofibrate intervention the expression of MMP-2 and MMP-9 protein levels drop.9.Enzyme spectrum method to detect further confirmed: fenofibrate can inhibition the enzyme activity of MMP-2 and MMP-9 in lung cancer A549 cells, and with the increase of drug concentration, the inhibition enhanced.Conclusion: 1. Fenofibrate can inhibit A549 cell growth, proliferation and induce its apoptosis.2. Fenofibrate can inhibit the invade athletic ability lung cancer A549 cells.3. Fenofibrate can inhibiting A549 lung cancer cell growth and function mechanism may be activated after PPARα increases the expression of TIMP-1 and TIMP-2, cut the expression of MMP-2 and MMP-9, and inhibition of MMP-2, MMP-9 enzyme activity.