Effects Of PARP-1Inhibitor PJ34Combined With Chemothearpy On Proliferation And Apoptosis Of Non-small Cell Lung Cancer

Objective To investigate the third-generation selective PARP-1inhibitor PJ34alone or combined with cisplatin, docetaxel on the effect of the cell proliferationinhibition and apoptosis of NSCLC cell lines A549, NCI-H460, so as to provide atheoretical basis for the PARP-1inhibitor PJ34clinical application,and expect thepossibility of the PARP inhibitors combining with the chemotherapy in the treatment ofNSCLC and provide new ideas for the treatment of lung cancer,for improving theprognosis of the patients with advanced lung cancer.Methods1. The human non-small cell lung cancer cell lines A549, NCI-H460cellswere cultured in vitro,and the anti-proliferative effect of PJ34, Cisplatin, Docetaxelalone or PJ34combined with Cisplatin/Docetaxel on A549and NCI-H460lines wasassessed by3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT).Drug-drug interacions were analyzed using combination index, The cells Morphologicalchanges were observed by Hoechst33342/PI fluorescent staining. The distributionchanges of the cell cycles were detect by Flow cytometry with PI stain, and theapoptosis rate of the cells was detect by Flow cytometry with AnnexinV-FITC/PI doublefluorescent staining.Results1.MTT colorimetry shows: compared with the control group, after variousconcentrations of PARP-1inhibitor PJ34separate treatment A549, H460cells, the cellproliferation was significantly inhibited, the difference was statistically significant (P<0.05), Dose-dependent growth inhibitory effects of PJ34were observed in NSCLC celllines A549,H460. the24h,48h, and72h IC50of PJ34for A549were (16.42±1.49)、(9.36±1.18)、(6.86±0.87)μg/ml, the IC50of PJ34for H460were respectively (13.67 ±1.94)、(8.33±1.16)、(5.38±0.43)μg/ml. the growth ofA549,H460cells significantlyslow down, and the cells decreased in the number and became shrinked in the cellmorphology compared with the control group,after PJ34treated.2. A549, H460cells were concurrently treated with PJ34and Cisplatin withconcentration of the1/8,1/4,1/2,1,2times IC50for72h, and the inhibition rate ofA549was respectively (13±1.12)、(31.25±2.4)、(55.32±4.32)、(83.31±5.23)、(93.78±2.8)%;and the cell inhibition rate of the H460cell line was (16.1±1.83)、(30.31±2.87)、(49±5.54)、(64.26±6.22)、(84.21±3.32)%. by Compusyn dose effect analysis softwareanalysing, PJ34combing with Cisplatin has a significant synergistic effect (CI <1).3. A549, H460cells were concurrently treated with the1/8,1/4,1/2,1,2times IC50concentration PJ34and Docetaxel for72h. The cell proliferation inhibition rate of theA549cells was (17.3±1.8)、(24.3±2.7)、(49.35±3.5）、(66.87±5.2)、(80.34±2.8)%, andcell inhibition rate of H460was (14.25±1.6)、(28.27±2.47)、(48.27±4.12)、(65.44±4.87)、(78.95±3.21)%, through Compusyn software analysis, PJ34combining withDocetaxel showed the antagonistic effect (CI>1)。4.Cell cycle analysis showed that the NSCLC A549, H460cells cycle was arrestedin G0/G1phase after PJ34treated for72h, and the number of cells in G0/G1phasebetween different concentrations was significantly more than the control group, and thecells in S phase decreased significantly compared with the control group. the differencewas statistically significant (P <0.01) in a dose-dependent way, and there was not muchdifference in the G2/M phase (P>0.05).5.Flow cytometry apoptosis detection showed that A549，H460cells apoptosisincreased significantly after PJ34treated when compared with the control group, andwhen PJ34combine with Cisplatin the cell apoptosis rates were much higher thanmonotherapy, and the difference was statistically significant (P <0.01).Conclusion1. Within a certain concentration range, PARP-1inhibitor PJ34has aninhibitory effect on the proliferation of the human NSCLC A549, NCI-H460cell lines, this effect is in time-and concentration-dependent manners.2The anti-proliferative activity of the PJ34for NSCLC A549, NCI-H460cells may be related to the cell cycle arrest and inducing apoptosis3.The synergistic effect was observed when PJ34was combined withCisplatin, however,(CI<1). However, the antagonistic effect was observed when PJ34was combined with Docetaxel(CI>1).