摘要
为研究肿瘤增殖基因Ki67反义多肽核酸(AS-PNAs)对人肾癌细胞的体内外抑制作用.将AS-PNAs(10.0μmol/L)转染人肾癌786-0细胞,采用免疫组化、Western印迹技术检测Ki67表达;3H-TdR掺入试验检测细胞增殖;免疫组化TUNEL法检测细胞凋亡。裸鼠移植瘤内注射AS-PNAs(10.0μmol/L)连续4d后,第3、6、12天处死小鼠,取瘤组织检测肿瘤体积、Ki67表达、细胞凋亡。结果发现,AS-PNAs处理组786-0细胞Ki67表达明显降低,3H-TdR掺入率明显降低,细胞凋亡率明显增加,与随机PNAs对照组比较差异均有显著性(P<0.01)。动物实验AS-PNAs处理组小鼠肿瘤体积明显缩小,Ki-67抗原表达明显降低,细胞凋亡明显增加,与对照组比较差异均有显著性(P<0.01)。Ki67基囚AS-PNAs在体外及动物体内均有抑制增殖、促进凋亡作用,是一种有前途的反义治疗药物。
To investigate the effect of the antisense peptide nucleic acid (AS-PNAs) of the tumor cell proliferation associated gene Ki67 on the proliferation and apoptosis of human renal carcinoma(HRC) cells, the HRC cell line 786-0 cells were transfected with AS-PNAs (10 μmol/L), and were implanted subcutaneously into BALB/c nude mice for 4 successive days to develop the implanted tumor model of HRC. The expression of the Ki67 gene in 786-0 cells was detected by immunohistochemical techniques and Western blotting, the proliferation of 786-0 cells was assayed by 3H-thymidine incorporation method, and the apoptosis of 786-0 cells was determined by TUNEL assay. It was found that the expression of Ki67 gene in 786-0 cells transfectd with AS-PNAs showed a significant reduction with a simultaneous reduction of proliferation of 786-0 cells as demonstrated by the decreased uptakes of 3H-thymidine, meanwhile, the apoptosis of 786-0 cells increased. The tumor volumes in the AS-PNAs treated group were significantly less than those in the control group. In addition, the expression of Ki67 gene in the AS-PNAs treated group was significantly lower, and the apoptosis was rather higher than those of the control group. These results indicate that AS-PNAs of the Ki67 gene shows definite effects on the proliferation and apoptosis of human renal carcinoma cells both in vitro and in vivo.
出处
《现代免疫学》
CAS
CSCD
北大核心
2005年第2期149-151,165,共4页
Current Immunology
基金
江苏省卫生厅医学科技发展基金资助项目(H2000153)江苏省教育厅资助项目(02KJD320030)
关键词
肾细胞癌
KI67基因
反义肽核酸
凋亡
裸鼠
renal cell carcinoma
Ki67 gene
antisense peptide nucleic acid
apoptosis
nude mouse
