利用SmartFlare技术进行胶质瘤细胞系U87中Twist1^(hi)亚群分选及其对顺铂耐药性研究

Twistl Confers Cisplatin Resistance inGlioma U87 Cells by Inhibiting Cisplatin-induced Apoptosis

目的利用特异性SmartFlare^(TM)RNA检测探针在人脑胶质瘤细胞系U87中分选Twist1^(hi)-U87细胞亚群,并研究其对顺铂的耐药性。方法用SmartFlare^(TM)RNA探针对Twist1^(hi)-U87细胞亚群进行分选。以未分选细胞作为对照。CCK8法检测Twist1^(hi)-U87亚群和亲代U87细胞对顺铂的化疗敏感性。流式细胞仪检测顺铂作用后细胞凋亡情况。结果根据细胞核内Twist1的表达量,流式细胞分选所得Twist1^(hi)-U87亚群约占亲代U87细胞的57.9%~69.8%。CCK8结果显示,加药后24h和48h,Twist1^(hi)-U87亚群对顺铂的耐药性均高于亲代U87细胞(P<0.05)。经顺铂处理后,Twist1^(hi)-U87细胞凋亡率(14.07%±0.96%)明显低于U87细胞(20.4%±1.29%)(P<0.05)。结论 Twist1基因高表达可以抑制胶质瘤细胞凋亡,进而降低顺铂的杀伤作用。

Objective Twist1 is a transcription factor that is highly expressed in many human cancers and its expression is related to chemotherapy resistance. In this study, we aimed to enrich for cells with a high-level expression of Twistl from human glioma cell line U87 and investigate their sensitivity to the chemotherapeutic agent cisplatin. Methods Twistlhi-u87 subsets were sorted through flow cytometry using Twistl specific SmartFlareTMRNA probe. Unsorted U87 cells were served as control. Different concentrations of cisplatin were administered to the cells. Cell viability was measured by CCK-8 assay and apoptosis was detected by flow cytometry. Results The percentage of Twistlhi-u87 subpopulations in unsorted parental cells was 57.9% - 69.8%. CCK-8 assay showed that Twistlhi-u87 subsets were more resistant to cisplatin than U87 cells at 24h, 48h （P 〈0.05）. In addition, Twistlhi-u87 subsets had a lower rate of apoptosis than did the parental U87 cells （ 14.07% ± 0.96% vs 20.4% ± 1.29% ） （ P 〈 0.05 ）. Conclusion Twistl is involved in cisplain resistance by inhibiting cisplatin-induced apoptosis.