MDA-7/IL-24基因与化疗药物联合应用对食管癌细胞产生协同抑制作用  

Enhanced cytotoxicity of chemotherapeutic agents in combination with adenoviruses expressing MDA-7 /IL-24 to esophageal carcinoma cells in vitro

作　　者：吴雅琼[1] 张明增[2] 刘北辰[2] 周凤灵 高玉环[2] 马光宇[2] 

机构地区：[1]河北医科大学第四医院心血管内科,河北石家庄050011 [2]河北医科大学第四医院东院区血液内科,河北石家庄050035

出　　处：《中国肿瘤生物治疗杂志》2014年第3期275-281,共7页Chinese Journal of Cancer Biotherapy

基　　金：河北省医学科学研究重点课题计划(No.20120125;No.20120128)~~

摘　　要：目的:探讨人黑素瘤分化相关基因7(melanoma differentiation-associated gene-7,MDA-7)/白介素-24(IL-24)基因对食管癌细胞的抑制作用及其与化疗药物的协同抗瘤作用。方法:RT-PCR法检测人食管癌细胞株TE-11和YES-5中MDA-7/IL-24受体IL-20R1、IL-20R2和IL-22R1的表达水平。用携带MDA-7/IL-24基因的重组人复制缺陷腺病毒Ad-MDA-7感染TE-11和YES-5细胞,Ad-LacZ为对照腺病毒,人成纤维细胞株OUMS-24为对照细胞。MTT法检测感染细胞抑制率,Western blotting法检测感染前后细胞中的MDA-7水平。化疗药物5-氟尿嘧啶(5-FU)、顺铂(CDDP)、丝裂霉素(MMC)和足叶乙甙(VP-16)分别与Ad-MDA-7联合作用于食管癌细胞株,MTT法检测单独或联合应用对食管癌细胞的抑制作用,流式细胞术检测AdMDA-7与化疗药单独或联合应用后食管癌细胞周期的变化,Western blotting检测Ad-MDA-7与化疗药联合作用后细胞凋亡和增殖的相关分子的变化。结果:TE-11和YES-5细胞均表达3种IL-24受体。Ad-MDA-7感染后,两种食管癌细胞中均有MDA-7蛋白表达,同时细胞均被剂量依赖性地抑制生长,Ad-MDA-7达3×104VP/细胞时TE-11细胞抑制率超过80%、YES-5细胞超过50%;同剂量Ad-LacZ对细胞无抑制作用,成纤维细胞OUMS-24被Ad-MDA-7感染后没有发生明显细胞抑制。AdMDA-7分别与5-FU、CDDP、MMC和VP-16联合应用后,与单独应用相比产生了更强的抗肿瘤协同效应。Ad-MDA-7与5-FU联合应用诱导细胞更多停滞在S和G2/M期,subG1期细胞比例明显增加。与单用5-FU相比,联合应用时Ad-MDA-7诱导了更多的细胞凋亡相关蛋白cleaved caspase-8、-9、-3的表达,增加了Akt的磷酸化,但降低了IκB-α的表达水平。结论:MDA-7/IL-24与化疗药联合应用于食管癌细胞,产生了更强的抗肿瘤协同效应,为临床化疗和基因治疗的联合应用提供新选择。Objective： To evaluate the synergic antitumor effects of chemotherapeutic agents and melanoma differentiation-associated gene-7-expressing adenoviruses（ Ad-MDA-7） in esophageal carcinoma cells in vitro. Methods： Human esophageal carcinoma TE-11 and YES-5 cells and human fibroblasts（ control） underwent Ad-MDA-7 infection and chemotherapy,either each alone or in combination. Changes in mRNA levels of the IL-24 receptor complexes before and after treatment were assessed by RT-PCR. Cell viability was determined by MTT assays. Cell cycle progression was analyzed by flow cytometry. Results： Transcripts for IL-24 receptor complex components,IL-20R2,IL-20R1 and IL-22R1,were detected in both TE-11 and YES-5 cells but only IL-20R2 mRNA was detected in fibroblasts. TE-11 and YES-5 cells were susceptible were to Ad-MDA-7-mediated cytotoxicity in a dose-dependent manner; at 3 × 104VP / cell,cytotoxicity was 〉 80% and 〉 50%,respectively,in TE-11 and YES 5 cells. In contrast,fibroblasts were resistant to Ad-mad-7. The cytotoxicity of 5-fluorouracil,cisplatin,mitomycin C or etoposide,each in combination with Ad-MDA-7 infection was significantly higher than that of these therapeutic agents and Ad-MDA-7,each alone. Increases in G 2 / M-phase and S-phase arrests were observed in cells,respectively,infected with Ad-MDA-7 and treated with 5-FU. The combination of Ad-mad-7 and 5-FU augmented sub-G1 populations. Compared with 5-FU alone,the combination regimen resulted in increases in caspase-8,-9,-3 expression and Akt phosphorylation and a decrease in IκB-α level. Conclusion： These data collectively suggest that adenovirus delivery of melanoma differentiation-associated gene-7 may enhance the sensitivity of esophageal carcinoma cells to chemotherapeutic agents through Akt activation.

关 键 词：食管癌 腺病毒 MDA-7 IL-24 基因治疗 化学治疗 

分 类 号：R735.1[医药卫生—肿瘤] R730.54[医药卫生—临床医学]