摘要
目的:观察c-myc反义寡核苷酸(ASODN)对胃癌MKN-45细胞株的生物学影响。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)评价对细胞增殖的影响,免疫细胞化学ABC方法检测转染后c-myc蛋白的表达;流式细胞仪(FCM)观察细胞凋亡;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基因反义寡核苷酸转染MKN-45细胞后,ASODN在0.25～4μmol/L的浓度范围内对MKN-45细胞均有不同程度的增殖抑制作用,作用48h后抑制率为11.2%～23.6%,且呈一定的剂量依赖性;FCM分析结果显示,转染后能诱导胃癌细胞凋亡,G0/G1期细胞增多;免疫细胞化学显示,c-myc蛋白水平明显降低,阳性表达率为(64.9±5.68)%;动物实验显示,ASODN可以抑制荷瘤小鼠肿瘤细胞的生长,肿瘤生长抑制率达41.5%。结论:c-myc基因反义寡核苷酸能明显抑制胃癌MKN-45细胞增殖、诱导细胞凋亡和下调c-myc蛋白水平。
OBJECTIVE, To observe the biological effect of c-myc antisense oligonucleotides （ASODN） on human gastric cancer MKN-45 cells. METHODS： The transfection on MKN-45 cells of c-myc antisense oligodeoxynucleotide was mediated by liposome. The cell proliferation was measured by MTT assay. The expression of c myc protein was measured by immunocytochemistry after the transfection. The apoptosis of cells was detected by flow cytometry and fluorescence microscopr. The tumor models were constructed of nude mice transplanted with human gastric cancer MKN 45 cells and the observations were made on tumor suppression rate and tumor volume. RESULTS.. After sealing c myc gene with ASODN, the proliferation of MKN45 cells was inhibited markedly and the inhibition rates treated with 0. 25 . FCM analysis 4 μmol/L ASODN were from 11.2% to 23.6%.showed that there appeared an obvious ap optosis peak after the transfection. The immunocytochemistry staining showed that c-myc protein expression was suppressed significantly（64.9 ± 5.68）%. The tumor volume decreased obvi ously and the rate of tumor suppression reached 41.5%0 in vivo experiment. CONCLUSION： Liposome-mediated c-myc ASODN can evidently inhibit cell proliferation,down-regulate c-myc protein expression, and induce apoptosis of MKN45 cells.
出处
《中华肿瘤防治杂志》
CAS
2007年第24期1860-1863,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
甘肃省中青年科技基金资助项目(3YS061-A25-025)
