QIN Hu, ZHENG Yi-tong, LIU Wen, MAIMAITI Aierpati, LI Ya-bin, WANG Yong-xin
Objective To investigate the effects of quinolinic phosphoribosyltransferase (QPRT) on the proliferation, migration and invasion of glioma cells, and to preliminarily elucidate its potential molecular regulatory mechanisms.Methods Stable QPRT knockdown and overexpression glioma cell lines (U87MG and U251MG) were established using lentiviral transduction. Western blotting was performed to detect the relative expression of QPRT protein and key enzymes involved in the nicotinamide adenine dinucleotide (NAD) biosynthesis pathway, including nicotinate phosphoribosyltransferase 1 (NAPRT1), nicotinamide phosphoribosyltransferase (NAMPT), nicotinamide mononucleotide adenylyltransferase 1 (NMNAT1), and NAD synthetase 1 (NADSYN1). Cell proliferation was assessed by CCK-8 assay, while cell migration and invasion were evaluated using wound healing assay and Transwell migration and invasion assays.Results Western blotting showed that, compared with knockdown control group, in U87MG cells, QPRT knockdown significantly increased NAPRT1 (t=3.431,P=0.027) and NMNAT1 (t=4.045,P=0.016) expression, and decreased NADSYN1 expression (t=29.715,P=0.000). Compared with overexpression control group, QPRT overexpression decreased NAPRT1 (t=8.970,P=0.009) and NMNAT1 (t=13.290,P=0.000) expression, and increased NADSYN1 expression (t=2.886,P=0.045). In U251MG cells, QPRT knockdown increased NMNAT1 expression (t=2.808,P=0.038) and decreased NADSYN1 expression (t=3.651,P=0.022), whereas QPRT overexpression decreased NMNAT1 expression (t=14.798,P=0.000) and increased NADSYN1 expression (t=4.314,P=0.013). Functionally, compared with knockdown control group, QPRT knockdown significantly reduced cell proliferation (U87MG:t=13.910,P=0.000; U251MG:t=6.118,P=0.004), migration (U87MG:t=5.750,P=0.005; U251MG:t=4.094,P=0.015), and the number of cells penetrating the membrane (migration:t=3.566,P=0.007;t=6.329,P=0.003; invasion:t=7.820,P=0.000;t=11.928,P=0.000). Conversely, compared with overexpression control group, QPRT overexpression significantly enhanced cell proliferation (U87MG:t=2.883,P=0.045; U251MG:t=4.094,P=0.015), migration (U87MG:t=4.908,P=0.008; U251MG:t=17.503,P=0.000), and the number of cells penetrating the membrane (migration:t=12.818,P=0.000;t=6.426,P=0.003; invasion:t=11.605,P=0.000;t=18.852,P=0.000).Conclusions QPRT promotes glioma cell proliferation, migration and invasion by regulating the NAD+biosynthesis pathway, plays an important role in the malignant progression of glioma, and may serve as a potential therapeutic target.