Effect of proteasome inhibitor MG132 combined with cisplatin on apoptosis and expression of Caspase-3 and Caspase-8 protein in lung cancer cells

WAN Qian; FANG Zhenyu; MA Nanshan; DU Zhuo

doi:10.3969/j.issn.1671-6450.2016.10.017

Abstract：Objective To investigate the effect of proteasome inhibitor MG132 combined with cisplatin on the apopto-sis of lung cancer NCI-H1299 cells and the expression of Caspase-3 and Caspase-8 protein.Methods Lung cancer NCI-H1299 cell line in vitro primary and subculture, will develop good cells into 4 groups: control group ( phosphate buffer was added 3 ml), cisplatin group (group D,added cisplatin 20 mg/L), M group (joined MG 132 2.5μmol /L), D +M group ( added cisplatin 20 mg /L +MG132 2.5μmol /L) .All groups were incubated 30 min.The lung cancer cell growth inhi-bition of each groups were measured with MTT colorimetric assay.The lung cancer cell morphology of each groups were ob-served with Cha Jixi gemcitabine.Lung cancer cells were measured by flow cytometry.The levels of Caspase-3, Caspase-8 protein were applicated with Western blot method.Results The results of MTT showed that M group, D+M group and NCI lung cancer cell proliferation rate of H1299 cells was significantly higher than that of C group and D group ( P <0.05), and D+M group of lung cancer cell proliferation rate was higher than that of M group ( P >0.05);after 48 h group, apoptosis of M cells in D+M group was higher than that of C group and D group (NCI-H1299 lung cancer cell culture P <0.05), and cell apoptosis of D+lung cancer in M group was higher than that of group M;Western blot showed that M group and D+M group the expression of Caspase-3 and Caspase-8 protein level was lower than that of C group and D group ( P <0.05). Conclusion The combination of MG132 and cisplatin can inhibit the apoptosis of lung cancer NCI-H1299 cells induced by TGF.The possible mechanism is that MG132 can effectively down regulate the expression of Caspase 3 and Caspase 8 protein.