Gao Xiao-kang, Li Qing, Wang He, Yang Bo, Yuan Jian-lin, Shao Guo-xing, Kang Fu-xia
Department of Urologic Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
Zhonghua Zhong Liu Za Zhi. 2005 Aug;27(8):468-70.
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.
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. Apoptosis index (AI) and proliferation index (PI) were examined by flow cytometry.
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 epirubicin 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).
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 epirubicin.
探讨靶向生存素的反义寡核苷酸(ASODN)对肾癌细胞系786 - O凋亡、增殖的影响及其对表柔比星敏感性的增强作用。
设计并构建靶向生存素的ASODN。将培养的细胞分为6组:对照组、脂质体组、正义寡核苷酸(SODN)组、600 nmol/L ASODN组、600 nmol/L ASODN联合表柔比星组。转染24 h后,收集培养的细胞进行下一步检测。通过透射电子显微镜观察细胞形态变化。采用免疫组织化学方法检测生存素蛋白。通过流式细胞术检测凋亡指数(AI)和增殖指数(PI)。
ASODN转染组观察到细胞形态异常。与对照组、脂质体组和SODN组相比,ASODN组生存素的表达显著降低。ASODN组的AI显著高于其他组。ASODN组的PI显著低于其他组。ASODN联合表柔比星组的PI为(35.7±1.67)%,而脂质体组或SODN组联合表柔比星时分别为(9.3±0.34)%或(8.5±0.21)%。与对照组相比,ASODN组增强凋亡的作用最强(P<0.05),而SODN未引起统计学显著变化(P>0.05)。
生存素ASODN可下调肾透明细胞癌细胞系786 - O中生存素蛋白的表达。靶向生存素的ASODN诱导786 - O细胞凋亡并抑制其增殖。抑制生存素增强了786 - O对表柔比星的敏感性。