c-myc反义寡核苷酸联合5-Fu对胃癌的体外及裸鼠体内抗肿瘤作用  

In vitro and in vivo synergetic antitumor effect of c-myc antisense oligodeoxynucleotides combined with 5-fluorouracil

作　　者：蔡辉[1] 王晓鹏[1] 苏河[1] 李荣范[1] 马云涛[1] 李振军[1] 

机构地区：[1]甘肃省人民医院普外科,甘肃兰州730000

出　　处：《第四军医大学学报》2007年第20期1830-1833,共4页Journal of the Fourth Military Medical University

基　　金：甘肃省中青年科技基金(3YS061-A25-025)

摘　　要：目的:探讨c-myc反义寡核苷酸(ASODN)联合5-Fu对胃癌MKN-45细胞株的体内、外抗肿瘤作用.方法:采用脂质体介导c-myc反义寡核苷酸转染人胃癌细胞系,用四甲基偶氮唑盐法(MTT法)评价c-mycASODN联合5-Fu对胃癌细胞增殖的影响;流式细胞仪(FCM)检测细胞凋亡率和细胞周期;RT-PCR及免疫组化方法检测c-myc基因表达变化;建立荷瘤小鼠模型,通过皮下瘤体内注射药物观察肿瘤体积和抑瘤率的变化.结果:c-myc反义寡核苷酸联合5-Fu可显著抑制细胞增殖,细胞凋亡率(23.4±1.9)%,较单用c-myc反义寡核苷酸(10.6±1.1)%和5-Fu(12.5±1.0)%的抑制作用更为显著(P<0.01);RT-PCR及免疫组化显示ASODN和5-Fu均使c-mycmRNA和蛋白表达下降;体内实验显示c-myc反义寡核苷酸联合5-Fu可明显减少肿瘤体积,抑制肿瘤生长,抑瘤率达46.7%;较单一治疗组明显(P<0.05).结论:c-myc基因反义寡核苷酸联合5-Fu能明显抑制胃癌MKN-45细胞增殖、诱导细胞凋亡和下调c-myc蛋白水平;且可有效抑制人胃癌裸鼠皮下肿瘤的生长.AIM： To investigate the antitumor effect of c-myc antisense oligonucleotides combined with 5-fluorouracil in vitro and in vivo. METHODS： The transfection of MKN-45 cells with c-myc antisense oligodeoxynucleotide was mediated by liposome, Cell proliferation was obseved by MTT assay. The apoptotic rate and cell cycle were examined by flow cytometry （FCM）, The expression of c-myc gene was detected by the method of immunohistochemistry and RT-PCR at the mRNA and protein level respectively. Nude mice tumor models were conducted with transplanting human gastric cancer MKN 45 cells and tumor suppression rate and tumor volume were then evaluated. RESULTS： The apoptotic rate of the group by c-myc antisense oligonucleotides combined with 5-fluorouracil reached （ 23.4 ± 1. 9 ） %, which was higher markedly than the rate of c-myc antisense oligonucleotides （ 10.6 ±1.1 ） % or 5-fluorouracil （ 12.5 ± 1. 0 ） % seperately （ P 〈0.01 ）. The tumor volume and the rate of tumor suppression of the combined group reached 46.7% in vivo experiment, which was significant compared to the separate group （ P 〈 0.05 ）. The results of immunohistochemical and RT-PCR assays showed that c-myc antisense oligonucleotides and 5-Fu could decrease the expression of c-myc gene. CONCLUSION： C-myc antisense oligonucleotides combined with 5-fluorouracil can inhibit cell proliferation, down-regulate c-myc gene expression, induce the apoptosis of MKN45 cells and effectively suppress the growth rate of gastric cancer cells in subcutaneous tumor. It can provide a new option to treat the gastric carcinoma.

关 键 词：基因 myc 5-氟尿嘧啶 寡核苷酸类 反义 胃肿瘤 体外/体内 

分 类 号：R73-36[医药卫生—肿瘤]