IL-24重组载体的构建及其联合化疗药物对SKOV3细胞中TUBB3与ERCC-1基因表达的影响  

Construction of Recombinant Vector on IL-24 and Its Effect in Combination with Chemotherapy Drugs for TUBB3 and ERCC-1 Expression in SKOV3 Cells

作　　者：祝贺[1] 杜珍武[1] 范丽梅[1] 高海成[2] 崔满华[1] 

机构地区：[1]吉林大学第二医院,长春130021 [2]吉林大学药学院,长春130021

出　　处：《高等学校化学学报》2015年第5期927-931,共5页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:81272875);吉林省科技厅项目(批准号:20140204023YY)资助~~

摘　　要：通过基因重组技术将白介素24(IL-24)基因片段分别克隆成pc DNA3.0-OSP-1-IL-24和pc DNA3.0-IL-24载体,利用逆转录PCR(RT-PCR)测定2种载体的表达.将2种化疗药物紫杉醇(PTX)与顺铂(DDP)分别作用于正常SKOV3细胞及pc DNA3.0,pc DNA3.0-IL-24,pc DNA3.0-OSP-1和pc DNA3.0-OSP-1-IL-24稳定转染SKOV3细胞(卵巢癌细胞株),通过噻唑蓝(MTT)法检测化疗药物联合IL-24基因对细胞的杀伤效果及细胞的增殖能力,应用实时荧光定量PCR技术检测人β-微管蛋白3(TUBB3)与核苷酸切除修复交叉互补基因位1(ERCC1)基因在各组细胞中的表达.RT-PCR检测结果显示,通过基因重组技术克隆了pc DNA3.0-IL-24与pc DNA3.0-OSP-1-IL-24载体,IL-24基因能提高SKOV3细胞对化疗药物顺铂和紫杉醇的敏感性.其联合化疗药物对肿瘤药物的IC50值分别下降了10倍和6倍,TUBB3和ERCC1基因在IL-24基因转染的SKOV3细胞内的表达水平与在正常SKOV3细胞内相比,分别下降4倍和10倍多.上述结果表明,IL-24联合化疗药物可明显下调TUBB3与ERCC1基因的表达,该实验为临床治疗卵巢癌提供了重要的理论依据.Gene fragments of IL-24 were cloned into pc DNA3.0-OSP-1-IL-24 and pc DNA3.0-IL-24 vector using gene recombination technology,the expression of two vectors was measured by RT-PCR methods.The two chemotherapy drugs of paclitaxel and cisplatin were applied into normal SKOV3 cells,stable transfected SKOV3 cells of pc DNA3.0,Pc DNA3.0-IL-24,pc DNA3.0-OSP-1 and pc DNA3.0-OSP-1-IL-24,respectively.The cell killing effect and proliferation ability of IL-24 in combination with chemotherapy drugs were detected by 3-（4,5）-dimethylthiahiahiazo-（z-yl）-2,5-di-phenytetrazolium bromide（MTT） in SKOV3 cells.Gene expression of TUBB3 and ERCC-1 were detected by real-time fluorescent quantitative polymerase chain reaction（PCR） technology in cell of each group.RT-PCR results showed that the pc DNA3.0-IL-24 and pc DNA3.0-OSP-1-IL-24 vector were cloned successfully by gene recombination technology.IL-24 gene can increase the sensitivity of SKOV3 cells to chemotherapeutic drug of cisplatin and paclitaxel.Its combination with chemotherapy drugs for IC50 of tumor drugs decreased 10 times and 6 times,respectively.Gene expression of TUBB3 gene and ERCC-1 decreased 4 times and 10 times,respectively.These results suggested that IL-24 in combination with chemotherapy drugs could reduce gene expression of TUBB3 and ERCC-1.This experiment provided an important theory basis for the clinical treatment of ovarian cancer.

关 键 词：IL-24基因 顺铂 紫杉醇 SKOV3细胞 

分 类 号：O629[理学—有机化学] R737.31[理学—化学]