姜黄素调控MicroRNA-1246及对膀胱癌T24细胞的放疗增敏机制探究

目的:研究在膀胱癌化疗中使用姜黄素的增敏功能原理。方法:在T24细胞经过姜黄素及放射处理后,探讨分析其细胞活力(细胞增殖试验试剂盒)、microRNA表达(miRNA芯片测序)、集落形成、凋亡(AnnexinV-F)分析、miR-1246及p53 mRNA及蛋白质(Westernblot)表达、ITC/7-AAD流式细胞计数(ITC/7-AAD)。结果:通过测试,发现17个异常microRNA表达,细胞在经过姜黄素处理后,T24细胞活力相较于常规组明显下降,并出现浓度依赖性的现象。在T24细胞中,miR-1246相较于人膀胱上皮永生化细胞(SV-HUC-1细胞)表达明显较高。姜黄素的浓度较高时,T24细胞miR-1246的表达会受其影响并明显下降。研究组使用浓度为20μg/mL的姜黄素,并结合放疗处理的方式,与常规组相比,研究组在控制miR-1246的表达、集落形成及细胞活力方面,效果更为显著,T24细胞比例会因转染antagomiR-1246而增加,细胞死亡现象一般发生于转染antagomiR-NC细胞时期。结论:膀胱癌细胞中,姜黄素和放射治疗都可促进microRNA-1246表达下调,在对肿瘤的治疗中可以使姜黄素及放疗结合,共同产生抗肿瘤作用。

A study on regulate and control microRNA-1246 andradiosensitization mechanism on bladder cancer T24 cells with curcumin

Objective:To study the radiosensitization mechanism on bladder cancer T24 cells with curcumin.Methods:After given curcumin and radiation,the cell viability(by cell proliferation test kit),microRNA expression(by miRNA chip sequencing),colony formation,apoptosis(by AnnexinV-F)of T24 cell,miR-1246 and p53 mRNA and protein(Westernblot)expression,ITC/7-AAD flow cytometry were analyzed.Results:Through testing,17 abnormal microRNA expressions were found.After the cells were treated with curcumin,the cell viability of T24 was significantly decreased compared with the conventional group,and the concentrationdependent phenomenon was appeared.In T24 cells,the expression of miR-1246 was significantly higher than SV-HUC-1 cells.When the concentration of curcumin is higher,the expression of miR-1246 in T24 cells will be affected and significantly decreased.After given 20μg/mL curcumin radiation and in the research group,there were more effective in controlling miR-1246 expression,colony formation and cell viability.T24 The proportion of cells will increase due to transfection of antagomiR-1246,and cell death generally occurs during the period of transfection of antagomiR-NC cells.Conclusion:In bladder cancer cells,curcumin and radiation therapy can promote the downregulation of microRNA-1246.In the treatment of tumors,curcumin and radiation can be combined to produce antitumor effects.