Franskevych D V, Grynyuk I I, Prylutska S V, Pasichnyk G V, Petukhov D M, Drobot L B, Matyshevska O P, Ritter U
Taras Shevchenko National University of Kyiv, Kyiv 01601, Ukraine.
Palladin Institute of Biochemistry, NAS of Ukraine, Kyiv 01011, Ukraine.
Exp Oncol. 2016 Jun;38(2):89-93.
To estimate the combined action of C60 fullerene and light irradiation on viability of L1210 leukemic cells, nitric oxide (NO) generation, p38 mitogen-activated protein kinase (MAPK) activity and cell cycle distribution.
Cell viability was assessed by MTT test. Light-emitting diode lamp (λ = 410-700 nm, 2.45 J/cm(2) ) was used for C60 fullerene photoexcitation. Nitrite level and NO-synthase activity were measured by Griess reaction and by conversion of L-arginine to L-citrulline, respectively. p38 MAPK activity was assessed by Western blot analysis. Cell cycle distribution was determined by flow cytometry.
It was shown that light irradiation of C60 fullerene-treated L1210 cells was accompanied by 55% decrease of their viability at 48 h of culture. Nitrite level measured as an index of reactive NO generation was increased at the early period after C60 fullerene photoexcitation due to activation of both constitutive and inducible NO-synthase isoforms. The simultaneous activation of p38 MAPK was detected. Accumulation of L1210 cells in sub-G1 phase of cell cycle was observed after C60 fullerene photoexcitation.
Photoexcited C60 fullerene exerts cytotoxic effect, at least in part, through triggering production of reactive NO species and activation of p38 kinase apoptotic pathways in L1210 leukemic cells.
评估C60富勒烯与光照联合作用对L1210白血病细胞活力、一氧化氮（NO）生成、p38丝裂原活化蛋白激酶（MAPK）活性及细胞周期分布的影响。
采用MTT法评估细胞活力。使用发光二极管灯（λ = 410 - 700 nm，2.45 J/cm²）对C60富勒烯进行光激发。分别通过格里斯反应和将L - 精氨酸转化为L - 瓜氨酸来测量亚硝酸盐水平和NO合酶活性。通过蛋白质免疫印迹分析评估p38 MAPK活性。通过流式细胞术确定细胞周期分布。
结果表明，在培养48小时时，光照处理的C60富勒烯处理的L1210细胞活力下降了55%。作为活性NO生成指标测量的亚硝酸盐水平在C60富勒烯光激发后的早期由于组成型和诱导型NO合酶亚型的激活而增加。检测到p38 MAPK的同时激活。在C60富勒烯光激发后观察到L1210细胞在细胞周期的亚G1期积累。
光激发的C60富勒烯至少部分地通过触发活性NO物质的产生和激活L1210白血病细胞中的p38激酶凋亡途径发挥细胞毒性作用。
