Ki67反义核酸对裸鼠人肾癌细胞移植瘤生长及凋亡的影响

Effects of antisense oligodeoxynucleotides against Ki67 mRNA on the growth and apoptosis of implanted human renal carcinoma cells in nude mice

目的 观察Ki67基因反义寡核苷酸(ASODNs)对裸鼠人肾癌细胞移植瘤生长及凋亡的影响。方法 BALB／C-nu裸鼠接种入肾癌786-0细胞,治疗组瘤体注射ASODNs(10 mmol／g),连续4 d。对照组注射RPMI 1640培养基。药物处理后第3、6、12天每次处死小鼠4只,取瘤组织检测肿瘤体积;免疫组织化学、Western blot检测Ki67表达;脱氧核苷酸末端转移酶介导的缺口末端标记(TUNEL)法检测凋亡。结果ASODNs处理组小鼠肿瘤体积缩小,与对照组比较差异有显著性(P<0.01);肿瘤Ki-67抗原表达率、Ki-67蛋白定量均降低,与对照组比较差异均有显著性(P<0.01);肿瘤细胞凋亡率增加,与对照组比较差异均有显著性(P<0.01)。结论Ki67 ASODNs能有效抑制裸鼠入肾癌细胞移植瘤生长并促进其凋亡。

Objective To investigate the effects of antisense oligodeoxynucleotides against Ki67 gene (ASODNs) on the growth and apoptosis of implanted human renal carcinoma (HRC) cells in nude mice. Methods Human renal carcinoma 786-0 cells were implanted subcutaneously in BALB/C-nu nude mice to develop the implanted tumor model of HRC. In ASODNs treated group, ASODNs (10 nmol/100 μl) were directly injected into tumors in nude mouse every day for 4 days. In control group nude mice were treated with the same volume of RPMI 1640. Twelve nude mice were killed at 3rd, 6th, 12th day respectively after treatment in each group. The tumor sizes were measured and tumor volumes were calculated. The Ki67 expression of tumor was detected by immunohistochemical technique and Western blot method respectively. The apoptosis of tumor was detected by TUNEL assay. Results The tumor volume in ASODNs group was significantly less than that in the control group (P<0. 01) . The Ki67 positive expression rate in ASODNs group was significantly lower than in the control group (P< 0. 01). The expression level of Ki67 protein in ASODNs group was significantly lower than in the control group ( P < 0.01).The apoptosis rate in ASODNs group was significantly higher than in the control group (P< 0.01). Conclusion ASODNs against Ki67 gene can inhibit the growth and induce apoptosis of implanted HRC cells in nude mice by blocking Ki67 expression of HRC cells.