Mechanism of oxymatrine promoting the apoptosis of osteosarcoma MG63 cell line through mitophagy mediated by COX-2/PINK1/Parkin signaling pathway
OBJECTIVE To study the mechanism of oxymatrine inducing apoptosis of osteosarcoma MG63 cell line based on mitophagy mediated by cyclooxygenase-2(COX-2)/PTEN-induced putative kinase-1(PINK1)/Parkinson disease protein-2(Parkin)signaling pathway.METHODS MG63 cells were treated with 2.0,4.0,8.0 mg/mL oxymatrine and 6 μmol/L 5-fluorouracil,then the apoptotic rate,the expression of apoptosis-related proteins[B-cell lymphoma-2(Bcl-2),Bcl-2 related X protein(Bax)],the proportion of decrease in mitochondrial membrane potential,the level of mitophagy as well as the protein expressions of PINK1,Parkin,and microtubule-associated protein 1 light chain-3Ⅱ(LC3-Ⅱ)were detected.PINK1 small interfering RNA(siRNA)was adopted to intervene in the expression of PINK1,the cells were divided into control group,PINK1 siRNA group,oxymatrine group,and PINK1 siRNA+oxymatrine group;the protein expressions of PINK1,Parkin,and LC3-Ⅱ,the proportion of decrease in mitochondrial membrane potential(MMP)as well as apoptotic rate were detected.The lentivirus infection technique was used to overexpress COX-2,the cells were divided into control group,oxymatrine group,COX-2 group,and COX-2+oxymatrine group.The protein expressions of COX-2,PINK1,and Parkin,as well as the proportion of decrease in MMP were detected.RESULTS After being treated with oxymatrine,the apoptotic rate,the protein expressions of Bax,PINK1,Parkin,and LC3-Ⅱ,the level of mitophagy as well as the proportion of decrease in MMP were significantly increased(P＜0.05),while the protein expression of Bcl-2 was significantly decreased(P＜0.05).Compared with the oxymatrine group,the protein expressions of PINK1,Parkin,and LC3-Ⅱ,apoptotic rate and the proportion of decrease in MMP were significantly decreased in PINK1 siRNA+oxymatrine group(P＜0.05).Compared with the oxymatrine group,the protein expression of COX-2 in the COX-2+oxymatrine group was increased significantly(P＜0.05),while the protein expressions of PINK1 and Parkin as well as the proportion of decrease in MMP were decreased significantly(P＜0.05).CONCLUSIONS Oxymatrine can mediate the overactivity of mitophagy based on the PINK1/Parkin signaling pathway by inhibiting COX-2 expression,thus promoting the apoptosis of the MG63 osteosarcoma cell line.
国家科技期刊平台
NSTL
万方数据
维普
