靶向抑制Slug基因表达对BxPC-3细胞凋亡和对5-FU化疗敏感性影响的实验研究  

Effects of Slug gene silencing by siRNA on increasing sensitivity of pancreatic cancer cell line BxPC-3 to 5-FU-induced apoptosis

作　　者：张克君[1] 曹红诗[2] 欧树安[3] 王齐全[4] 魏小斌[5] 

机构地区：[1]青岛大学医学院附属医院普外二科,山东青岛266003 [2]青岛大学医学院附属医院肿瘤科,山东青岛266003 [3]海南医学院附属医院临床外科实验室,海南海口517010 [4]海口市人民医院消化内科,海南海口517000 [5]海口市人民医院中心实验室,海南海口517000

出　　处：《中华肿瘤防治杂志》2010年第12期919-922,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨siRNA干扰Slug基因表达对BxPC-3细胞增殖、凋亡和对5-FU化疗敏感性的影响。方法:用脂质体转染法将表达siRNA-Slug的真核表达载体pGenesil-1-Slug-siRNA(pSlug-siRNA)和空载体pGen-esil-1-Neg-siRNA(pNeg-siRNA)导入胰腺癌BxPC-3细胞,G418筛选阳性细胞,获得稳定转染的阳性克隆。将转染载体的BxPC-3阳性克隆细胞和未转染载体的BxPC-3细胞分别暴露于系列浓度(0.01～100μmol/L)的5-FU72h。RT-PCR和蛋白质印迹法检测不同组细胞经5-FU作用72h后的Slug表达;MTT法测定各组细胞生存率并计算IC50,Hoechst33258和PI双染色和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测细胞的凋亡。结果:实验组细胞内Slug基因被有效沉默;BxPC-3、Bx-PC-3/pSlug-siRNA和BxPC-3/pNeg-siRNA细胞的5-FUIC50分别为(11.0±2.1)、(1.5±0.4)和(9.7±1.3)μmol/L(P<0.05).BxPC-3/pSlug-siRNA细胞对5-FU作用的敏感性增加了7.33倍。5-FU以剂量依赖方式诱导BxPC-3细胞凋亡,但对BxPC-3/pSlug-siRNA细胞所诱导的凋亡比BxPC-3和BxPC-3/pNeg-siRNA更明显。结论:靶向抑制Slug基因表达促进胰腺癌BxPC-3细胞凋亡,并增强BxPC-3对5-FU的化疗敏感性。OBJECTIVE：To investigate whether Slug gene silencing by siRNA can increase the sensitivity of pancreatic cancer cells （PC） to 5-FU-induced apoptosis.METHODS：pSlug-siRNA and pNeg-siRNA were transfected into BxPC-3 cells by the lipofectamine 2000 methods,and the stable transfected clony was chosen through G418.BxPC-3 cells were divided into three groups：a transfected pSlug-siRNA group,transfected pNeg-siRNA group and control group without transfection BxPC-3 cells,and the three groups were treated with serial concentrations （0.01-100 μmol/L） of 5-FU.The MTT assay was used to observe the inhibitory actions of 5-FU on BxPC-3 cells,and the apoptosis was assessed by Hoechst 33258 and PI dye staining and TUNEL staining.The expression of Slug protein was detected by Western blotting and RT-PCR.RESULTS：The inhibitory action of cell growth was seen in the 3 groups’ cells dealing with 5-FU.It also promoted the occurrence of apoptosis.The IC50 of BxPC-3,BxPC-3/pSlug-siRNA and BxPC-3/pNeg-siRNA were （11.0±2.1）,（1.5±0.4） and （9.7±1.3） μmol/L （P〈0.05）.The sensitivity of BxPC-3/pSlug-siRNA to 5-FU was incveased by 7.33 times.5-FU inhibited the proliferation of BxPC-3 cells in a concentration-dependent manner.In pSlug-siRNA cells treated with 5-FU,the apoptosis rate increased greatly and the proliferation rate decreased compared with the control cells and pNeg-siRNA cells dealing with 5-FU.The expression of Slug was decreased greatly compared with pNeg-siRNA cells treated with 5-FU.CONCLUTION：Slug gene silencing by siRNA greatly enhances the sensitivity of BxPC-3 cells to 5-FU-induced apoptosis.

关 键 词：胰腺肿瘤/药物疗法 基因 slug 氟尿嘧啶 

分 类 号：R735.9[医药卫生—肿瘤]