姜黄素通过调控miR-1305表达影响胶质瘤细胞对顺铂的耐药性

Curcumin Affects Cisplatin Resistance of Glioma Cells by Regulating the Expression of miR-1305

目的探讨姜黄素对人胶质瘤细胞株(U251)/顺铂(cisplatin,DDP)耐药性的影响及其对微小RNA(miR)-1305的调控作用。方法以顺铂浓度递增和间歇大剂量冲击诱导相结合的方法建立U251/DDP耐药株。用不同剂量姜黄素和不同浓度顺铂处理U251/DDP耐药株,分为对照组、低剂量姜黄素组、中剂量姜黄素组、高剂量姜黄素组、DDP组、DDP+姜黄素组、DDP+miR-NC(阴性)组、DDP+miR-1305组、DDP+姜黄素+anti-miR-NC组和DDP+姜黄素+anti-miR-1305组。采用MTT法检测U251与U251/DDP增殖抑制率并计算顺铂IC50值;qRT-PCR法检测miR-1305表达量;MTT法和流式细胞术检测细胞增殖抑制率和凋亡率。结果U251/DDP细胞增殖抑制率明显低于U251(P<0.05),U251/DDP的顺铂IC50值明显高于U251(P<0.05);与对照组比较,不同剂量姜黄素组细胞增殖抑制率升高(均P<0.05)、顺铂IC50值降低(均P<0.05)、miR-1305表达水平升高(均P<0.05)。与DDP组比较,DDP+姜黄素组U251/DDP细胞增殖抑制率和凋亡率升高(均P<0.05);与DDP+miR-NC组比较,DDP+miR-1305组U251/DDP细胞增殖抑制率与凋亡率升高(均P<0.05);与DDP+姜黄素+anti-miR-NC组比较,DDP+姜黄素+anti-miR-1305组细胞增殖抑制率与凋亡率降低(均P<0.05)。结论姜黄素可通过上调miR-1305表达抑制细胞增殖和促进细胞凋亡,从而增强胶质瘤对顺铂治疗的敏感性。

Aim To explore the effect of curcumin on cisplatin resistance of glioma cells and its regulatory effect on microRNA(miR)-1305.Methods The cisplatin-resistant glioma cell line U251/DDP was established by a combination of cisplatin(DDP)concentration increase and intermittent highdose shock induction,and the MTT assay was used to detect the proliferation inhibition rate of human glioma cells U251 and U251/DDP,and calculate the cisplatin IC50 value.U251/DDP cells were treated with different doses of curcumin and different concentrations of cisplatin,and MTT assay was used to detect the proliferation inhibition rate of U251/DDP cells and calculate the cisplatin IC50 value.Using U251/DDP cells as research materials,the experimental groups were as follows:a control group,a lowdose curcumin group,a middle-dose curcumin group,a high-dose curcumin group,a DDP group,a DDP+curcumin group,a DDP+mi R-NC group,a DDP+mi R-1305 group,a DDP+curcumin+anti-mi R-NC group,and a DDP+curcumin+anti-mi R-1305 group.q RT-PCR method was used to detect the expression of mi R-1305.MTT experiment and flow cytometry were used to detect cell proliferation inhibition rate and apoptosis rate respectively.Results The proliferation inhibition rate of U251/DDP cells was significantly lower than that of U251 cells(P<0.05),and the IC50 value of cisplatin in U251/DDP cells was significantly higher than that of U251 cells(P<0.05).Compared with the control group,the inhibition rate of cell proliferation was increased(P<0.05),the IC50 value of cisplatin was decreased(P<0.05),and the expression level of mi R-1305 was increased(P<0.05).Compared with DDP group,the U251/DDP cell proliferation inhibition rate and apoptosis rate in DDP+curcumin group were increased(P<0.05).Compared with DDP+mi R-NC group,the U251/DDP cell proliferation inhibition rate and apoptosis rate in DDP+mi R-1305 group were increased(P<0.05).Compared with DDP+curcumin+anti-mi R-NC group,the cell proliferation inhibition rate and apoptosis rate in DDP+curcumin+anti-mi R-1305 group were decreased(P<0.05).Conclusion Curcumin could inhibit cell proliferation and promote cell apoptosis by up-regulating the expression of mi R-1305,thereby enhancing the cisplatin sensitivity of gliomas.