CAJ | 학술논문

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.MTT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RhoGDI2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RhoGDI2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、RhoGDI2、Maspin是潜在的结肠癌多药耐药相关基因.
목적 사선결장암다약내약상관기인,탐토결장암다약내약적궤제.방법 통과축보체증불뇨밀정(5-FU)농도부육결장암Lovo세포,건립내약세포주Lovo/5-FU.MTT법검측Lovo/5-FU세포대5-FU화순박(CDDP)적민감성.채용이유전영-질보련용기(2DE-MS)술비교Lovo화Lovo/5-FU량주세포간차이표체단백,병채용Western blot법감정사선단백.결과 여Lovo상비,Lovo/5-FU세포대5-FU화CDDP적IC.치분별승고31배화3배(균P＜0.01).통과2DE-MS사선득도,재내약주중CAP-G화RhoGDI2표체상조,6-PGL、DCI、Prdx-6화Maspin표체하조.Western blot검측현시,Lovo/5-FU세포중RhoGDI2화CAP-G교Lovo증고6.14배화2.98배(균P＜0.01),이Maspin칙명현하강지Lovo중적5.2％(P＜0.01).결론 결장암내약성적산생시다기인、다통로개변적결과.CAP-G、RhoGDI2、Maspin시잠재적결장암다약내약상관기인.
Objective To identify novel multi-drug resistance-related genes,and to explore the mechanisms of multi-drug resistance.Methods Multi-drug 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 dimersional electrophoresis plus mass spectrum(2-DE/MS) was used to identifv the differentially expressed protein between Lovo and Lovo/5-FU.The identified protein was then verified by Western blot analysis. Results The IC50 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 Multi-gene and multi-pathway are involved in the development of multi-drug resistance of colorectal cancer cells.CAP-G,RhoGDI2 and Maspin are potential multi-drug resistant genes.