Zhang Pei, Liu Fang, Gao Jiao, Ma Lin-Yan, Sun Xiao-Jin, Zheng Hai-Lun, Liu Hao, Zhao Su-Rong
School of Pharmacy, Bengbu Medical College/Anhui Provincial Engineering Technology Research Center of Biochemical Pharmaceuticals, Anhui Bengbu 233030, China.E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Jul 20;37(7):883-888. doi: 10.3969/j.issn.1673-4254.2017.07.05.
To investigate the effect of small interfering RNA (siRNA)-mediated silencing of monocarboxylate transporter 1 (MCT1) on the sensitivity of drug-resistant nasopharyngeal carcinoma HNE1/DDP cells to cisplatin (DDP)-induced apoptosis and explore the possible mechanism.
The expression of MCT1 was analyzed in HNE1 and HNE1/DDP cells and in HNE1/DDP cells transfected with siRNA using Western blot. MTT assay was used to assess the inhibitory effect of different concentrations of DDP alone or in combination with MCT1 siRNA on the proliferation of HNE1/DDP cells. The apoptosis of cells treated with MCT1 siRNA or/and DDP (8 µmol/L) was assessed using flow cytometry with PI staining, and the mitochondrial membrane potential was detected using JC-1 staining assay; the expressions of Mcl-1, Bak, Bcl-2, and Bax were analyzed using Western blotting.
HNE1/DDP cells showed a high expression of MCT1, and MCT1 silencing using siRNA significantly increased the sensitivity of HNE1/DDP cells to DDP (P<0.05) and partly reversed DDP resistance of the cells. MCT1 silencing enhanced the sensitivity of HNE1/DDP cells to DDP-induced apoptosis. Treatment of HNE1/DDP cells with MCT1 siRNA combined with 8 µmol/L DDP for 24 h resulted in an apoptotic rate of (51.23∓2.86)%, significantly higher than that in cells treated with MCT1 siRNA or DDP alone (P<0.05). The combined treatment also reduced the mitochondrial membrane potential, down-regulated the expression of Mcl-1 and Bcl-2, and up-regulated the expression of Bax in the DDP-resistant cells.
MCT1 siRNA can enhance the sensitivity of HNE1/DDP cells to DDP-induced apoptosis, the mechanism of which may involve the down-regulation of Mcl-1 and Bcl-2 and up-regulation of Bax expression.
探讨小干扰RNA（siRNA）介导的单羧酸转运体1（MCT1）沉默对耐药鼻咽癌HNE1/DDP细胞顺铂（DDP）诱导凋亡敏感性的影响，并探讨其可能机制。
采用蛋白质免疫印迹法分析HNE1和HNE1/DDP细胞以及转染siRNA的HNE1/DDP细胞中MCT1的表达。采用MTT法评估不同浓度DDP单独或联合MCT1 siRNA对HNE1/DDP细胞增殖的抑制作用。采用PI染色流式细胞术评估MCT1 siRNA或/和DDP（8 μmol/L）处理细胞的凋亡情况，采用JC-1染色法检测线粒体膜电位；采用蛋白质免疫印迹法分析Mcl-1、Bak、Bcl-2和Bax的表达。
HNE1/DDP细胞中MCT1表达较高，使用siRNA沉默MCT1可显著提高HNE1/DDP细胞对DDP的敏感性（P<0.05），并部分逆转细胞的DDP耐药性。沉默MCT1增强了HNE1/DDP细胞对DDP诱导凋亡的敏感性。MCT1 siRNA联合8 μmol/L DDP处理HNE1/DDP细胞24 h，凋亡率为（51.23±2.86）%，显著高于单独使用MCT1 siRNA或DDP处理的细胞（P<0.05）。联合处理还降低了耐药细胞的线粒体膜电位，下调了Mcl-1和Bcl-2的表达，上调了Bax的表达。
MCT1 siRNA可增强HNE1/DDP细胞对DDP诱导凋亡的敏感性，其机制可能与下调Mcl-1和Bcl-2以及上调Bax表达有关。
