Investigation Of Effect Of Fructus Viticis Total Flavonoids And Casticin On Induction Of Growth And Apoptosis In Lung Cancer A549Cell

Objective: To investigate the growth inhibitory and proapoptotic effects of humanlung cancer A549cells by treatment with Fructus Viticis total flavonoids(FVTF) andcasticin(CAS) that is an active constituent of FVTF. To investigate whether theirmechanisms are involved in down-regulate the protein expression of FoxM1.Methods: Human lung cancer A549cell line was cultured in vitro, and treated withvarious concentrations of CAS and FVTF. Colony formation on the plate was used toexamine the inhibitory effects of Casticin on the growth of A549cell. Flowcytometry(FCM) with propidium iodide(PI) staining was emploied to analyze the cellapoptosis rate and cell cycle. The cell apoptosis morphology was observed by AO/EBFluorescent staining. The expression level of FoxM1protein was analyzed by Westernblot analysis.Results:1.The CAS group was tested with concentrations of0.3,1,3,10,15,30μmol/L and the DDP group with concentration of3μg/mL. The colony inhibition ratesof A549cells were2.06±0.38%,19.51±2.61%,46.15±5.12%,58.16±7.76%,65.67±3.11%,71.86±2.90%and73.55±2.89%respectively. There was significantdifference compared to the solvent control group (P<0.05), when IC50was7.26μmol/L.2. The CAS group with concentration of10,30μmol/L and the DDP groupwith3μg/mL were observed under AO/EB Staining fluorescence microscope. Thetypical morphological changes of apoptotic cell was found in A549cells.3. Solventcontrol group: the CAS group with concentration of3,10,30μmol/L and the DDPgroup with3μg/mL, the apoptosis rates of A549cells were1.81±0.91%,3.26±2.08%,5.27±4.29%,10.64±5.40%and23.13±6.91%respectively, there was a significantdifference compared to the solvent control group (P<0.05).4. The expression ofintracellular FoxM1protein decreased in a concentration-and time-dependent manner.5. The FVTF group was tested with concentrations of0.3,1,3,10,15,30μmol/L andthe DDP group with concentration of3μg/mL. The colony inhibition rates of A549cells were3.72±0.39%,27.00±3.74%,40.41±1.72%,53.63±2.38%,67.78±3.23%,71.86±2.28%and72.25±2.33%respectively. There was a significant difference compared to the solvent control group (P<0.05), with IC50was16.02μmol/L;6. TheFVTF group(20,40μg/mL) and DDP of3μg/mL were observed under AO/EB Stainingfluorescence microscope. The typical morphological changes of apoptotic cell wasfound in A549cells.7. Solvent control group,10,20,40μg/mL of FVTF group and3μg/mL of DDP group was measured by PI staining flow cytometry. Cell apoptosisrates of A549cells were1.81±0.91%,4.35±0.91%,6.74±0.57%,8.67±0.58%and23.13±6.91%. There was a significant difference compared to the solvent controlgroup (P<0.05);8. A549cells treated by FVTF using western blot analysis, there wasno obvious change in FoxM1protein expression.Conclusion:1.Casticin can inhibit growth and induce apoptosis in lung cancer A549cell, in aconcentration-dependent manner.2.Casticin can a concentration-dependent down-regulate the protein expression ofFoxM1.3.FVTF can inhibit growth and induce apoptosis in lung cancer A549, in aconcentration-depeendent manner.4.The mechanism underlying FVTF’s effect of induction of cell growth inhibition andapoptosis did not associate with alternation of FoxM1protein expression.