CRLP与DNA修复相关性的初步研究  

Preliminary study on relativity between CRLP and DNA repair

作　　者：汪朝晖[1] 覃文新[1] 潘志美[1] 万大方[1] 顾健人[1] 

机构地区：[1]上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海200032

出　　处：《肿瘤》2003年第1期14-18,共5页Tumor

基　　金：国家重点基础研究发展规划"973"资助项目(G1998051004)

摘　　要：目的探讨细胞经紫外处理后CRLP与DNA修复的相关性。方法将插入反义CRLP的pcDNA3.1/V5-His质粒用脂质体介导法转染BEL7402肝癌细胞,G418筛选获得的细胞用15J/m2的紫外作用,4h后用Annexin V—EGFP试剂处理细胞,荧光激活细胞分选法测定细胞凋亡比例。另外用RT-PCR和Northern杂交的方法检测了肿瘤细胞株和多组织中的CRLP的表达。结果转染反义CRLP质粒的细胞株经紫外处理后,统计学处理表明凋亡比率显著高于对照;该基因在所检测的肿瘤细胞株和各类组织中均有表达。结论CRLP基因的表达受到抑制以后可能通过影响细胞DNA修复能力降低细胞抗紫外杀伤的能力。Objective To explore the relativity between CRLP and DNA repair in cell treated with ultraviolet ray. Methods CRLP and antisense CRLP were inserted into pcDNA3.1/V5-His plasmid and transfected into BEL7402 hepatocellular carcinoma cell line by using liposome transfection reagent. The G418 screened cells were exposed to 15J/m2 ultraviolet ray and treated with Annexin V-EGFP reagent four hours later, then the ratio of apoptotic cell was determined by fluorescence-activated cell sorting. In addition, RT-PCR and Northern blot were used to detect the expression of CRLP in tumor cell lines and multiple tissues. Results The apoptotical ratio of antisense CRLP transfected cell line was dramatically higher than that in control group. CRLP expressed in all the detected tumor cell lines and tissues. Conclusion Inhibition of CRLP may reduce the DNA repair ability through blocking DNA repairing complex forming.

关 键 词：DNA修复 荧光激活细胞分选法 CRLP基因 相关性 肿瘤细胞 

分 类 号：R730.21[医药卫生—肿瘤]