结肠癌多药耐药性相关基因筛选与鉴定  

Screening and identification of genes associated with multi-drug resistance in colonic cancer

作　　者：李建芳[1] 郑重[1] 俞焙秦[1] 瞿颖[1] 朱正纲[1] 刘炳亚[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院外科上海消化外科研究所上海市胃肿瘤重点实验室,200025

出　　处：《中华胃肠外科杂志》2012年第4期388-391,共4页Chinese Journal of Gastrointestinal Surgery

基　　金：国家自然科学基金（30872476）;上海市重点学科-外科学（S30204）：上海市重大科技专项（09DZ1950100）：上海市科委重点课题（10JC1411100）

摘　　要：目的筛选结肠癌多药耐药相关基因，探讨结肠癌多药耐药的机制。方法通过逐步递增氟尿嘧啶（5-FU）浓度孵育结肠癌Lovo细胞，建立耐药细胞株Lovo／5-FU。M1Tr法检测Lovo／5-FU细胞对5-FU和顺铂（CDDP）的敏感性。采用二维电泳-质谱联用技（2DE—MS）术比较Lovo和Lovo／5-FU两株细胞间差异表达蛋白，并采用Western blot法鉴定筛选蛋白。结果与Lovo相比，Lovo／5-FU细胞对5-Fu和CDDP的IC。值分别升高31倍和3倍（均P〈0.01）。通过2DE—MS筛选得到。在耐药株中CAP—G和RhoGD12表达上调，6-PGL、DCI、Prdx-6和Maspin表达下调。Western blot检测显示．Lovo／5-FU细胞中RhoGD12和CAP-G较Lovo增高6．14倍和2．98倍（均P〈0．01）．而Maspin则明显下降至Lovo中的5．2％（P〈0．01）。结论结肠癌耐药性的产生是多基因、多通路改变的结果。CAP—G、RhoGD12、Masoin是潜在的结肠癌多药耐药相关基因。Objective To identify novel multi-drug resistance-related genes, and to explore the mechanisms of multi-drug resistance. Methods Multi-dmg resistant cell line Lovo/5-FU was established by incubation with increasing dose of 5-FU. The sensitivity to 5-FU and cis-diaminodichloroplatinum （CDDP） was measured by MTT assay. Two dimensional electrophoresis plus mass spectrum（2-DE/MS） was used to identify the differentially expressed protein between Lovo and Lovo/5-FU. The identified protein was then verified by Western blot analysis. Results The ICSO concentrations of Lovo/5-FU to 5-Fu and CDDP were increased by 31 and 3 times, compared with Lovo （both P〈0.01）. 2DE-MS showed that CAP-G and RhoGDI2 were up-regulated, whereas 6-PGL, DCI, Prdx-6 and Maspin were down-regulated in Lovo/5-FU. Western blot analysis confirmed that the expression levels of RhoGDI2 and CAP-G in Lovo/5-FU were increased by 6.14 and 2.98 fold respectively （both P〈0.01）, whereas Maspin was decreased to 5.2% of Lovo （P〈0.01）. Conlusions Muhi-gene and multi-pathway are involved in the development of muhi-drug resistance of colorectal cancer cells. CAP-G, RhoGD12 and Maspin are potential multi-drug resistant genes.

关 键 词：结肠肿瘤 多药耐药 氟尿嘧啶 蛋白组学 

分 类 号：R735.35[医药卫生—肿瘤]