摘要
目的探讨microRNA-125b(miR-125b)在人视网膜母细胞瘤细胞中多药耐药的作用及其机制。设计实验研究。研究对象人视网膜母细胞瘤SO-RB50细胞。方法用RT-PCR方法检测miR-125b视网膜母细胞瘤细胞株SO-RB50和耐药细胞株SO-Rb50/VCR中的表达变化;化学合成的miR-125b过表达(miR-125mimic组)和抑制载体(miR-125inhibitor组)转染SO-Rb50细胞株,用MTT法和Annexin V-FITC法检测在药物长春新碱、依托泊苷和卡铂,依次作用上述转染细胞后,细胞增生力和细胞凋亡的变化;用蛋白印迹法检测miR-125b过表达和抑制表达后细胞株SO-RB50中MAGE-A/P53蛋白的表达变化。主要指标细胞存活率和细胞凋亡率。结果SO-Rb50/VCR组与SO-RB50组相比,miR-125b的表达显著增高(P=0.002);长春新碱、依托泊苷和卡铂依次作用于转染后的SO-RB50细胞株后,miR-125mimic组与miR-125inhibitor组相比,细胞存活率显著增高(P=0.000),细胞凋亡率显著下降(P=0.000),P53蛋白表达水平显著下降(P=0.001),MAGE-A蛋白表达水平显著增高(P=0.004)。结论在SO-RB50细胞中,下调miR-125b后提高肿瘤细胞对化疗药物敏感性,且miR-125b可能是通过MAGE-A/P53通路调控视网膜母细胞瘤多药耐药性。
Objective To explore the effects of microRNA-125 b(miR-125b) on the retinoblastoma multidrug resistance and to study its molecular mechanism of miR-125b on chemotherapy sensitivity.Design Experimental study.Participants The retinablastoma cell lines SO-RB50.Methods The expression of miR-125b was detected by real-time PCR in SO-RB50 cell line and SO-RB50/VCR cell line.The synthesized miR-125b mimic(miR-125 mimic group) and miR-125b inhibitor(miR-125 inhibitor group) was transfected into retinoblastoma SO-RB50 cell line.In addition,three chemotherapeutic drugs,including carboplatin,etoposide and vincristine,were used to treat the transfected SO-RB50 cell line respectively,in order to evaluate the sensitivity of RB cells.The cell proliferation and apoptosis were measured by MTT and Annexin V-FITC.The protein expression level of MGEA-A and P53 was detected in the transfected SO-RB50 cell line by Western blot.Main Outcome Measures The percentage of cell survival and the percentage of apoptotic cells to total cells.Results MiR-125 b was significantly up-regulated(P=0.000)in the SO-RB50/VCR cell line compared to that in the SO-RB50 cell line(P=0.000).It apparently promotes RB cell proliferation and suppresses cell apoptosis(P=0.000) in the miR-125 mimic group compared with that in the miR-125 inhibitor group treated by carboplatin,etoposide and vincristine respectively(P=0.000).The protein expression level of MAGE-A was higher significantly in the miR-125 mimic group than in the miR-125 inhibitor group(P=0.004).M eanwhile,the protein expression level of P53 was lower significantly in the miR-125 mimic group than in the miR-125 inhibitor group(P=0.001).Conclusion The SO-RB50 cell line inhibited the expression of miR-125b was sensitive to carboplatin.etoposide and vincristine,and furthermore miR-125b/MAGE-A/p53 axis may be conducive to enhancing the efficacies of chemotherapeutic treatments for RB.
作者
白淑玮
毕春潮
张广良
邵娟
许治国
Bai Shuwei;Bi Chunchao;Zhang Guangliang;Shao Juan;Xu Zhiguo(Department of Ophthalmology,Xi'an Fourth Hospital,Xi'an 710004,China;Department of Ophthalmology,Tongliao City Hospital,Tongliao 028000,Inner Mongolia,China)
出处
《眼科》
CAS
2020年第6期470-475,共6页
Ophthalmology in China
