新的肿瘤耐药相关基因ERGIC-53的研究

目的:研究ERGIC-53基因表达与肿瘤细胞耐药的关系,寻找肿瘤耐药逆转可能的新靶点。方法:采用Northern Blot检测ERGIC-53基因在K562和K562/A02中的表达差异;应用逆转录聚合酶链反应(RT-PCR)检测11种肿瘤敏感及耐药细胞系中ERGIC-53基因的表达;设计合成针对ERGIC-53基因的siRNAs,用脂质体法转染K562和K562/A02细胞,用MTT比色法和流式细胞仪检测其干扰耐药细胞中ERGIC-53基因表达后的细胞对阿霉素的敏感性。结果:ERGIC-53在K562/A02细胞中的表达明显高于亲代K562细胞(P<0.05);6种耐药细胞株中ERGIC-53基因表达的平均值约为亲代敏感细胞的2倍。siRNA转染组与对照组相比,K562/A02细胞对阿霉素的敏感性均有不同程度的提高,且在转染后72h时细胞内阿霉素积累接近其亲代敏感细胞水平。结论:ERGIC-53基因与肿瘤细胞耐药相关,明确该基因参与耐药肿瘤细胞表型的形成。

Study on A Novel Tumor-Related Drug Resistant Gene ERGIC-53

Objective:To investigate the role of ERGIC-53 gene in tumor drug resistance,and provide a new target for circumventing tumor drug resistance in the future.Methods:The different expression levels of ERGIC-53 between K562/A02 and its parental K562 cell line were determined by Northern blot.The method of RT-PCR was used to detect the expression level of ERGIC-53 in 11 kinds of tumor cell lines,including sensitive and drug resistant cell lines.The siRNA targeted to ERGIC-53 were transfected into K562/A02 by Lipofectamine 2000.The expressions of ERGIC-53 in those transfected cells were analyzed by RT-PCR,and the sensitivity to doxocubicin of K562/A02 was investigated by MTT assay and FACS.Re-sults:The expression level ERGIC -53 was significantly increased,about two -fold higher,in K562/A02 than that in its parental K562 cell line.The siRNA targeted to ERGIC-53 decreased the ERGIC-53 expression level,and significantly in-creased doxocubicin accumulation in K562/A02,and sensitize ERGIC-53-expressing K562/A02 resistance to doxocubicin.Conclusion:ERGIC-53 may play an important role in tumor drug resistance,and further studies are needed for a clear un-derstanding of the molecular mechanism of this novel drug resistant-related gene.