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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 study the inhibitory effect of Ki-67 antisense polypeptide nucleic acid (AS-PNAs) on human renal cell carcinoma cells in vitro and in vivo, AS-PNAs (10.0μmol / L) were transfected into human renal carcinoma 786-0 cells using immunohistochemistry Western blotting was used to detect the expression of Ki67. 3H-TdR incorporation assay was used to detect cell proliferation. Immunohistochemical TUNEL method was used to detect apoptosis. Mice were sacrificed on the 3rd, 6th and 12th day after AS-PNAs injection (10.0μmol / L) for 4 days in vivo. The tumor volume, Ki67 expression and apoptosis were detected. The results showed that the Ki67 expression of 786-0 cells in AS-PNAs treated group was significantly decreased, the 3H-TdR incorporation rate was significantly decreased, and the apoptosis rate was significantly increased, which was significantly different from the control group (P <0.01). Compared with the control group, the tumor volume of AS-PNAs treatment group was significantly reduced, the expression of Ki-67 antigen was significantly decreased, and the apoptosis rate of AS-PNAs group was significantly increased (P <0.01). Ki-67-based AS-PNAs, as a promising antisense drug, inhibit proliferation and promote apoptosis in vitro and in vivo.