Reversion of multidrug resistance in human glioma by RNA interference
Reversion of multidrug resistance in human glioma by RNA interference
复制标题
RNA干扰逆转人神经胶质瘤的多药耐药性
DOI：
10.1179/174313208x297869
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发表时间：
2008-07
影响因子：
1.9
通讯作者：
He, Yue
中科院分区：
文献类型：
--
作者：
Zhang, Yazhuo;Sun, Meizhen;Zhao, Peng;He, Yue
摘要 目的：探讨体内短发夹载体构建能否使人胶质瘤细胞系BT325产生RNA干扰（RNAi）双链体并逆转MDR1基因的表达。方法：根据GeneBank MDR1序列构建3个62nt寡核苷酸片段（shRNA），并克隆至逆转录病毒载体。将这些载体通过Lipofectamine 2000与增强型绿色荧光蛋白（EGFP）共转染直接转染至BT325细胞后，通过实时PCR（RT-PCR）、Northern blot和Western blot分析，通过mRNA和P-糖蛋白（P-gp）水平的变化检测MDR1基因沉默效应。为了评估对阿霉素（DOX）和长春新碱（VCR）的多药耐药性，通过细胞计数试剂盒8进行细胞增殖测定并计算IC50。结果：RNAi质粒载体构建成功。 Northern blot转染48小时后基因沉默最强； Western blot分析表明P-gp表达量降低至12.9%、30.3%和4.8%。化疗敏感性实验表明，转染细胞可增加DOX和VCR的敏感性。从IC50值来看，BT325细胞对药物的敏感性明显增加。序列特异性RNAi可以抑制胶质瘤细胞系的MDR1 mRNA和P-gp表达。它可能逆转多药耐药表型，这可能为人类神经胶质瘤的治疗提供有希望的治疗方式。
Abstract Objective: To explore whether the vector construction of short hairpin in vivo could make human glioma cell line BT325 produce RNA interference (RNAi) duplexes and reverse the expression of the MDR1 gene. Methods: Three 62nt oligonucleotide fragments (shRNA) were constructed according to GeneBank MDR1 sequence and were cloned to the retrovirus-delivered vectors. After these vectors were transfected directly to the BT325 cell by Lipofectamine 2000 with enhanced green fluorescent protein (EGFP) co-transfection, the MDR1 gene silence effects were detected by the changing levels of mRNA and P-glycoprotein (P-gp) including real-time PCR (RT-PCR), Northern blot and Western blot analysis. For assessing multidrug resistance against doxorubicin (DOX) and vincristine (VCR), cell proliferation assays were performed by cell counting kit-8 and IC50 was calculated. Results: The RNAi plasmid vectors were constructed successfully. The gene silence became the strongest after 48 hour transfection from Northern blot; Western blot analysis demonstrated that P-gp expression reduced to 12.9, 30.3 and 4.8%. The chemosensitivity assays showed that the transfected cell could increase the sensitivity of DOX and VCR. Based on the value of IC50, BT325 cells increased sensitivity to drugs obviously. The sequence specific RNAi could inhibit MDR1 mRNA and P-gp expression of the glioma cell line. And it may reverse multidrug resistance phenotype, which may provide promising therapeutic modalities in the treatment of human glioma.
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影响因子：
64.8
作者：
Mello, CC;Conte, D
通讯作者：
Conte, D
DOI：
--
发表时间：
2000
期刊：
--
影响因子：
--
作者：
杨贤志
通讯作者：
杨贤志
影响因子：
3.5
作者：
Nieth, C;Priebsch, A;Lage, H
通讯作者：
Lage, H
影响因子：
6
作者：
Chao Liu;I. A. Qureshi;Xun-Jie Ding;Yi-Fei Shan;Y. Huang;Yi Xie;Mei-Rong Ji
通讯作者：
Chao Liu;I. A. Qureshi;Xun-Jie Ding;Yi-Fei Shan;Y. Huang;Yi Xie;Mei-Rong Ji
DOI：
--
发表时间：
--
期刊：
--
影响因子：
--
作者：
B. Davidson;H. Paulson
通讯作者：
B. Davidson;H. Paulson