抑制氯通道CLCN2基因表达对顺铂诱导胶质瘤U251细胞损伤的影响

Effect of CLCN2 antisense oligonucleotide on U251 cells injury by cisplatin

目的:探讨使用特异性CLCN2反义寡核苷酸抑制CLCN2基因表达对顺铂诱导的神经胶质瘤U251细胞损伤的影响。方法:U251细胞按处理方法不同分4组,对照组(转染无义寡核苷酸)、转染CLCN2反义寡核苷酸组、顺铂组(无义寡核苷酸与顺铂联用)、CLCN2反义寡核苷酸与顺铂联用组。应用MTT法检测U251细胞生存率;RT-PCR检测CLCN2、cyclinD1 mRNA表达;TUNEL法检测细胞凋亡率。结果:与对照组相比,转染CLCN2反义寡核苷酸组、顺铂组和CLCN2反义寡核苷酸与顺铂联用组U251细胞生存率、CLCN2 mRNA表达降低,凋亡细胞数增加;与顺铂组相比,CLCN2反义寡核苷酸与顺铂联用组细胞生存率(P<0.05)、CLCN2、cyclinD1 mRNA表达降低,凋亡细胞数增加(P<0.01);与CLCN2反义寡核苷酸组相比,CLCN2反义寡核苷酸与顺铂联用组CLCN2 mRNA表达明显降低(P<0.01)。结论:①转染CLCN2反义寡核苷酸能有效抑制CLCN2 mRNA表达;②抑制CLCN2 mRNA表达能促进顺铂对U251细胞的损伤作用;③CLCN2基因表达降低参与顺铂对U251细胞损伤作用。

AIM： To investigate the effect of chloride channel CLCN2 antisense oligonucleotide on the cell injury of malignant U251 glioma cells induced by cisplatin （DDP）. METHODS： The experiment was divided into 4 groups： control group （nonsense oligonucleotide） , CLCN2 antisense oligonucleotide group, DDP group （DDP ＋ nonsense oligonucleotide）, DDP ＋ CLCN2 antisense oligonucleotide group. The viability of U251 cells was measured by MTT assay, CLCN2 mRNA level was determined by RT - PCR, cell apoptosis was measured by TUNEL assay. RESULTS： Compared to the control group, the cell viability, CLCN2 and cyclin D1 mRNA decreased in CLCN2 antisense oligonucleotide group, DDP treated group and CLCN2 antisense oligonucleotide with DDP treated group, cells apoptosis increased. Compared to DDP group, the cell viability （P 〈 0.05 ） and CLCN2 mRNA decreased in CLCN2 antisense oligonucleotide with DDP treated group, and cells apoptosis increased （P 〈 0.01 ）. Compared to CLCN2 antisense oligonucleotide group, CLCN2 mRNA significantly decreased （P 〈 0.01 ） in CLCN2 antisense oligonucleotide with DDP treated group. CONCLUSION： CLCN2 antisense oligonucleotide inhibits the expression of CLCN2 mRNA in U251 cells. Inhibition of CLCN2 mRNA facilitates the cell injury of U251 cells induced by DDP. The decrease in CLCN2 mRNA is involved in the mechanism of cell injury by DDP.