应用染色体显微切割技术筛选人小细胞肺癌多药耐药相关基因的实验研究  

Experimental research of screening multidrug-resistant genes in human small cell lung cancer by chromosome microdissection

作　　者：戢福云[1] 钱频[2] 吴国明[1] 李福祥[1] 郑世珍[1] 陈维中[1] 陈琰[1] 钱桂生[1] 

机构地区：[1]第三军医大学附属新桥医院全军呼吸内科研究所,重庆400037 [2]第三军医大学附属新桥医院超声科,重庆400037

出　　处：《中华肺部疾病杂志（电子版）》2010年第3期30-33,共4页Chinese Journal of Lung Diseases(Electronic Edition)

摘　　要：目的应用染色体显微切割技术筛选人小细胞肺癌多药耐药基因。方法在倒置显微镜下,显微切割人小细胞肺癌(small cell lung cancer,SCLC)多药耐药(multidrug-resistance,MDR)细胞系NCI-H446/CDDP特异性畸变染色体并构建该染色体片段DNA微文库,然后采用菌落斑点杂交、反Northern Blot及Northern Blot等方法,筛选人SCLC MDR相关基因。结果筛选出25个在耐药细胞中上调表达的基因或DNA片段。在已测序的20个DNA序列中,3个分别为人类2号和5号染色体BAC片段DNA序列,在全长库中未检索到其对应的相似序列,可能代表了某个基因或者该序列位于基因变异丰富的3’端,而无法查到与其它物种基因的同源性。其它17个序列与已知基因存在95%～100%的同源性。已知基因包括硫氧还蛋白、Bcl-2、TRAF、神经酰胺等基因。结论多个基因可能构成网络调控系统,参与人SCLC MDR相关基因的形成,但其具体的调控机制,则仍需大量的研究探索。Objective To screen multidrug-resistant genes in human small cell lung cancer by chromosome microdissection. Methods After the microdissection of specifically abnormal chromosomal fragments of NCI-H446/CDDP cells and construction of micro-DNA library, Dot blot, reverse Northern Blot and Northern Blot were used to screen and identify MDR-related genes. Results Totally, there were 25 DNA fragments or genes which were found to be up-regulated in NCI-446/CDDP cells. Among 20 fragments sequenced, 3 fragments were chromosome BAC sequences respectively and no homologs were found in GenBank. We thought the three sequences maybe belong to the 3 ＇ ends of certain genes where exited highly variable sequence and it was difficult to identify their homolog in GenBank. Other 17 sequences were found to be similar to several known genes with 95%-100% similarity. The known genes include Trx,Bcl-2,TRAF and Ceramide. Conclusion Many genes were involved in the development of MDR of human SCLC and the genes came into being a complex network which regulates the MDR. While, so far, the regulation mechanism was still unknown and many researches were need to be done.

关 键 词：SCLC 多药耐药 染色体显微切割 NORTHERN BLOT 

分 类 号：R563[医药卫生—呼吸系统]