摘要
目的探讨MiRNA-122对肝癌耐药细胞株BEL-7402/5-FU的5-氟尿嘧啶敏感性的影响以及作用机制。方法构建miR-122及其阴性对照空载体稳定转染至BEL-7402/5-FU细胞中,Western blot检测未转染组,阴性载体转染组和miR-122转染组中与耐药相关的p53蛋白表达水平。用MTT法和流式细胞术检测三组细胞对5-氟尿嘧啶敏感性。结果 MiR-122转染组与未转染组,阴性载体转染组比较,野生型p53蛋白表达水平升高。流式细胞术检测,miR-122转染组的细胞凋亡率较未转染组和阴性载体转染组高。结论 miR-122能上调野生型p53蛋白表达,增加BEL-7402/5-FU细胞对5-氟尿嘧啶敏感性,从而成为治疗肝癌耐药治疗的一个方法。
Objective To investigate the influence and mechanism of miR-122 confer 5-FU sensitivity in BEL- 7402/5-FU cells. Methods MiR-122 and negative miRNA expression vectors were constructed and stably transfected into BEL-7402/5-FU cells. Western Blotting was used to detect Wild-type p53 （ wt-p53 ） protein expression. Drug sensitivity of the cells to 5-fluorouracil （5-FU） were analyzed with MTF and flow cytometry. Results Compared with negative miR- NA transfectants or untreated cells, protein expression level of wt-p53 in stable miR-122 transfectants were decreased. Flow cytometry results demonstrated that apoptosis rate increased in miR-122 transfected cells, compared with negative miRNA or untreated ceils. Conclution MiR-122 can specifically up-regulate the expression of wt-p53 in BEL-7402/5-FU cells, which increased ceils spontaneous apoptosis and sensitize cells to 5-FU. Therefore, miR-122 may be used as a potential therapy agent against human hepatoblastoma.
出处
《中南医学科学杂志》
CAS
2011年第6期640-642,658,共4页
Medical Science Journal of Central South China
