论文部分内容阅读
目的左卡尼汀(levocarnitine,L-CNT)在改善由化疗药物引起的心脏毒性和周围神经毒性方面有一定的作用。本实验旨在研究联合应用时,L-CNT对多西他赛(docetaxel,DTX)抑制非小细胞肺癌(non-small cell lung cancer,NSCLC)GLC-82细胞增殖作用的影响,初步探讨其作用机制。方法 MTT法检测不同浓度L-CNT(0.625、1.25、2.5、5、10、20、40和80μg/mL)和DTX(0.01、0.1、1、5、10、20、40和80μg/mL)分别对GLC-82细胞增殖作用的影响,倒置显微镜观察其形态的变化。以终浓度为1.25、20、80μg/mL L-CNT分别和DTX(9.301μg/mL)联合,同时处理GLC-82细胞24h,MTT法检测其对细胞增殖作用的影响,流式细胞术检测细胞凋亡的变化,蛋白质印迹法检测Bcl-2、Bax和ANT1蛋白表达的情况。结果与阴性对照组相比,L-CNT(1.25、20和80μg/mL)对GLC-82细胞作用24h后,促进增殖率分别为(14.52±0.73)%、(27.03±1.21)%和(19.77±0.65)%,F=42.538,P=0.001;48h后分别为(11.37±0.13)%、(19.56±1.01)%和(14.89±0.51)%,F=52.119,P=0.001。与DTX(27.319μg/mL)单药组相比,L-CNT(1.25、20和80μg/mL)、DTX联合用药组的细胞增殖率由(49.49±2.52)%增加至(52.64±2.43)%、(59.44±3.20)%和(53.15±2.37)%,F=7.412,P=0.011。与DTX(9.301μg/mL)单药组相比,L-CNT(1.25、20和80μg/mL)、DTX联合用药组GLC-82细胞密度增大,细胞凋亡率由(24.67±1.95)%分别降低至(21.90±1.98)%、(14.38±2.35)%和(22.42±1.43)%,F=15.697,P=0.001。L-CNT、DTX联合作用后,抑凋亡蛋白Bcl-2表达增多,促凋亡蛋白Bax表达减少,同时ANT1蛋白表达降低。结论 L-CNT和DTX联合应用对GLC-82细胞增殖与凋亡有一定的影响,主要通过下调Bax和上调Bcl-2蛋白的表达以及影响线粒体膜通透性转换孔的开放和关闭,减弱DTX对细胞凋亡的诱导作用。
OBJECTIVE: Levocarnitine (L-CNT) plays a role in the improvement of cardiotoxicity and peripheral neurotoxicity induced by chemotherapy drugs. The purpose of this study was to investigate the effects of L-CNT on the proliferation of non-small cell lung cancer (NSCLC) GLC-82 cells treated with docetaxel (DTX) mechanism. Methods MTT assay was used to detect the expression of L-CNT (0.625,1.25,2.5,5,10,20,40 and 80μg / mL) and DTX (0.01,0.1,1,5,10,20,40 and 80μg / mL) On the proliferation of GLC-82 cells, the morphological changes were observed by inverted microscope. GLC-82 cells were treated with L-CNT at 1.25,20,80μg / mL and DTX (9.301μg / mL) respectively for 24 hours. The effects of L-CNT on cell proliferation were detected by MTT assay. Flow cytometry The changes of apoptosis and protein expression of Bcl-2, Bax and ANT1 were detected by Western blotting. Results Compared with the negative control group, the proliferation rates of GLC-82 cells treated with L-CNTs (1.25, 20 and 80 μg / mL) for 24 h were (14.52 ± 0.73)%, (27.03 ± 1.21)% and ± 0.65%, F = 42.538, P = 0.001 respectively; (11.37 ± 0.13)%, (19.56 ± 1.01)% and (14.89 ± 0.51)%, F = 52.119, P = 0.001 after 48h. Compared with DTX alone (27.319μg / mL), the cell proliferation rate increased from (49.49 ± 2.52)% to (52.64 ± 2.43)% in L-CNTs (1.25,20 and 80μg / mL) , (59.44 ± 3.20)% and (53.15 ± 2.37)% respectively, F = 7.412 and P = 0.011. Compared with DTX group (9.301μg / mL), the cell density of GLC-82 in L-CNTs (1.25, 20 and 80μg / mL) and DTX group increased, the apoptotic rate increased from (24.67 ± 1.95) (21.90 ± 1.98)%, (14.38 ± 2.35)% and (22.42 ± 1.43)% respectively, F = 15.697, P = 0.001, respectively. After the combination of L-CNT and DTX, the expression of Bcl-2 and Bax decreased, while the expression of ANT1 decreased. Conclusions The combination of L-CNT and DTX can affect the proliferation and apoptosis of GLC-82 cells by down-regulating the expression of Bcl-2 protein and up-regulating the expression of Bcl-2 and opening and closing of mitochondrial membrane permeability transition pore, Induction of apoptosis.