摘要
目的研究survivin反义寡核苷酸(ASODN)转染对肾透明细胞癌786O细胞的survivin蛋白表达、细胞凋亡、增殖的影响,及其对表阿霉素诱导细胞凋亡的作用。方法设计并合成特异性靶向survivin的ASODN及正义寡核苷酸(SODN),将其转入786O细胞。设空白对照组、脂质体对照组、SODN组和600nmol/LASODN组,处理24h后收获各组细胞。透射电子显微镜观察细胞形态变化,免疫组化法检测各组细胞survivin表达情况,流式细胞术检测各组细胞增殖和凋亡指数。结果ASODN组的细胞呈典型凋亡的形态学改变,而对照组细胞生长良好;ASODN组细胞survivin表达减弱,凋亡指数明显升高(P<0.05);转染ASODN24h后,表阿霉素诱导786O细胞凋亡的作用明显增强。结论survivinASODN能下调survivin蛋白表达,诱导肾透明细胞癌786O细胞凋亡,抑制786O细胞增殖,并增强表阿霉素诱导的细胞凋亡。
Objective To investigate the effect of antisense oligonucleotide （ASODN） targeting survivin on the apoptosis and proliferation of renal cancer cell line 786-O and enhancement of its sensitivity to epirubicin. Methods ASODN targeting survivin was designed and constructed. Cultured cells were divided into 6 groups： control group, liposome group, sense oligonucleotide （SODN） group, 600 nmol/L ASODN group, and 600 nmol/L ASODN combined with epirubicin group. After transfected for 24 h, cultured cells were harvested to carry on the next tests. Cell morphological changes were examined by transmission electron microscopy. Survivin protein was detected by immunohistochemical method. Apoptesis index （AI） and proliferation index （PI） were examined by flow cytometry. Results Morphological abnormalities of cells were observed in ASODN transfected groups. Expression of survivin in ASODN groups were significantly decreased compared with that in the control group, liposomes group and SODN group. AI of ASODN groups was significantly higher than that in other groups. PI of ASODN groups was significantly lower than that in other groups. The PI of ASODN combined with epiruhicin group was （35.7±1.67）%, but （9.3±0.34）% or （8.5±0.21）% in liposomes group or SODN group that had combined with epirubicin. The ASODN group achieved the strongest effects to enhance apoptosis in comparison with control group （ P 〈 0.05 ） , while SODN did not cause statistically significant change （ P 〉 0.05 ） . Conclusion The expression of survivin protein in the renal clear cell carcinoma cell line 786-O is downregulated by survivin ASODN. ASODN targeting survivin induces apoptosis and inhibits proliferation of 786-O cells. Inhibition of survivin enhances sensitivity of 786-O to epinthicin.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2005年第8期468-470,共3页
Chinese Journal of Oncology
