Ki67反义肽核酸、反义寡核酸对肾癌细胞系增殖及凋亡影响的研究

 目的 探讨肿瘤增殖相关基因Ki67反义肽核酸(PNAs)、反义寡核酸(ASODNs)对人肾癌细胞增殖及凋亡的调控。寻找肾癌反义治疗的合适药物。方法 将PNAs转染人肾癌786-0细胞系，采用免疫组化、Western blot技术检测Ki67表达，细胞生长曲线，³H-thymidine掺入试验检测肾癌细胞增殖，TUNEL法检测癌细胞凋亡。并与相同浓度的ASODNs进行对比。结果 PNAs处理组(10μmol／L)786-0细胞Ki67表达阳性率(％)(16．9±0．7)降低，Ki67蛋白(％)(42．1±2．2)降低，与ASODNs处理组(28．6±0．4)(83.6±1．4)比较差异有显著性(P<0．01，P<0．01)。PNAs处理组³H-thymidine掺入率(％)(20．7±1．5)减少，与ASODNs处理组(58．6±1．4)比较差异有显著性(P<0．01)。PNAs处理组凋亡细胞阳性率(％)(28．7±2．3)增加，与ASODNs处理组(13．8±1．0)比较差异有显著性(P<0．01)。结论 PNAs可在反义及反基因二个环节发挥抗肿瘤作用，与ASODNs相比，PNAs有更强的阻抑人肾癌Ki67基因表达、增殖及促进凋亡作用，是一种有前途的基因治疗药物。

Effects of Peptide Nucleic Acids Targeting Ki67 on the Proliferation and Apoptosis of Human Renal Carcinoma Cell Line

Objective 　To investigate the effects of peptide nucieic acid stargeting Ki67 gene ( PNAs) on the proliferation and apoptosis of human renal carcinoma cell line cells. Methods 　Human renal carcinoma cell line 786-0 cells were t reated with PNAs (10. 0μmol/ L) . The Ki67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot method respectively. The proliferation of 786-0 cells was studied by cell growth curves and 3>> H-thymidinuptake assay. The apoptosis of 786-0 cells was detected by TUNEL assay. The cont ral group was t reated with antisense oligonucleotides (ASODNs) of Ki67.Results 　The Ki67 experssion rate of 786-0 cells t reated by PNAs (16. 9 ±0. 7) was lower than that of ASODNs t reated group (28. 6 ±0. 4) ( P < 0. 01) . The Ki67 protein rate of 78620 cells t reated by PNAs (42. 1 ±2. 2) was lower than that of ASODNs t reated group (83. 6 ±1. 4) ( P < 0. 01) . The 3 >>H-thymidine incorporation rates of 78620 cells t reated by PNAs (20. 7 ±1. 5) was lower than that of ASODNs t reated group (58. 6 ±1. 4) ( P < 0. 01) . The apoptosis rate of 786-0 cells t reated by PNAs (28. 7 ±2. 3) was higher than that of ASODNs treated group (13. 8 ±1. 0) ( P < 0. 01) . Conclusion 　PNAs have both antisense and antigene effcct s. PNAs of Ki67 have more powerful effect s on the proliferation and apoptosis of human renal carcinoma cells than ASODNs of Ki67 gene.